Somali Movement

The initial opposing party to Barre's regime was conducted by the officers in the army after his defeated in Ogden war in 1977, some of them were sentenced and others escaped to Ethiopia and established the first opposing front called â€Å"Somali salvation Democratie Frof† (SSDF), as well as the immigrants of Ishaq tribe in England established â€Å"Somali National Movement† (SNM). Later in 1989, the United Somali Movement was formed by Hawiya tribe and controlled Mogadishu. The next year about (100) politicians signed a document demanding Barre to resign and appointing a transitional government. In 1991, the Somali Congress Forces (SCF) led by Mohammed Farah Aidid defeated the government forces, however Barre coerced to escape to â€Å"Gedo† near Kenyan borders. The ouster of Barre's government left behind a vacuum power which inspired the appetite of opposing groups to govern. An armed combat between (USC) commanders Mohammed Aidid and Ali Mahdi Mohammed to control of the capital. The fighting lasted for 100 days and the number of the victims was estimated about (30.000) thousands. On the meantime, the (SNM) proclaimed independence under the name of Somaliland. In 1992, a cease fire between Aidid and Ali Mehdi was a greed, and no one controlled the capital which amplify the gap between the north and the south. The United Nation Operation in Somalia I (UNOSOM I) was establish in 1992 to maintain ceasefire and support humanitarian relief efforts (Khaleji, 2007). Unfortunately, the ceasefire was ignored and fighting nonstop. In 1992, about (350.000) Somali died due to illness, hunger and civil war. The US established multinational forces to secure humanitarian processes and it became United Task Force (UNITF) after approving by Security Council. In March 1993, the United Nation operation in Somalia II (UNOSOM II) started to reconstruct Somali state and economy. They withdraw in 1995 after having incurred causalities. In 1995, the military combat between local parties became less intense. Aidid stated himself as a president of the Somalia without getting any recognition. His forces occupied Daidoa. In 1996, Ali Mehdi elected as a chairman of the United Somali Congress and Somali Calvation Alliance (USC//SSA). In the same year, Aidid died of his wounds. In 2000, Ali Mehdi lost votes to Abdulqasim Salad Hasssan. In the same year, the transitional national government (TNG) was established because of Somali National Arta Conference in Djibouti. In 2004, the Transitional Federal Government was established in Nairobi. In 2006, a clash between U.S backed militia leaders and Islamic Courts Union (ICU) which won the combat and controlled most of the Southern Somalia (Dresso,2009). Thus, Ethiopia entered Somalia as allies to (TFG) against (ICU). The next year president Abdullah control Mogadishu and US began strike against al-Qaida. In 2010, al-Shabaab declared a coalition with al-Qaida and attacked the capital. In the same year, the famine killed about 260,000 people. In 2012, the Federal Government of Somalia was established. In 2013, US recognized the Somali government for the first time since 1991. In the same year and the next, al-Shabaab conducted some attack operations against Kenya like what happened in November a mass killing in north-east Kenya. In 2016, the African Union leaders agreed to raise their troops and support their military to deteriorate al-Shabaab. In 2017, Mohammed Abdullah selected as a president of Somalia. In October 2017, bombing killed about 350 people in Mogadishu. In march 2018, about 18 citizens were killed and 22 others injured because of blast close to hotel in Mogadishu.

Writing and Colonial New England

Men were not responsible for anything that went on in the house back in that time. Married and divorced parents spent more time now with their children than 40 years ago. Children time for fathers Increased a lot more now than in the colonial times. Fathers weren't responsible for their children and women were obligated to do all house work. Response: This particular article took me by surprise because the fact that back in Colonial times fathers didn't really help around the house is upsetting and surprising.In my opinion, women and men are obligated to do the same and equal work as catheter. The Role of Men and Women In Colonial New England: Summary: Women and Men were forbidden to strike each other in the Colonial times. A man was forced to give bond if he was caught verbally abusing his wife. The duty of a husband was to go work and support his wife at all times. Women's property was forced to be given up to her husband once they were married and she was not allowed to work or ow n anything. In men's pollen women lacked strength for Intellectual exercise.Response: This article shocked me because the fact that men saw themselves as better than women is extremely degrading and unfair, women can do the same wings men can do The Role of Children In Colonial New England: Summary: Puritan parents were obligated to direct their children responsibly. Children who were too spoiled were sent to be treated by a master to become more obedient. Girls started learning house work as young as the age of 5. They had to learn how to cook and clean and do all the kinds of housework.I feel like with time writing exams you are so rushed to finish writing and outline that by the time you start your essay you Just go blank. To prevent that I read my articles more than once to completely understand it thoroughly and then I begin my essay. This really helps me in the long run and is good or completing and understanding my essay. Log 1 felt like I was prepared for the midterm. If I w ere to change anything I would read my articles a few more times next time to better understand them before my midterm.But generally I felt like I did a exceptional Job on my midterm and tried my hardest spending all the time I could to finish it. Writers Checklist: 1 . Does your idea draft respond fully to the assignment? Yes, it does. 2. Are your ideas organized the way you want? Yes, in my opinion, the ideas are organized how I want them to be. 3. Does your intro explain what the essay is about and what its repose is? My essays introduction introduces the topic and explains what the essay is going you be about. . Do you have a thesis that states your point or indicates the issue the essay will address? Yes, my thesis indicates the issue that my essay will address. 5. Do the body paragraphs each have a topic sentence? Do they develop the main points by giving specifics and examples to support these points? Yes. 6. Does your conclusion make one or more recommendations? Yes, my conc lusion makes at least one recommendation. 7. Yes, both my trusted friend and a classmate has reviewed my essay.

Diagnostic Control Systems: Implementing Intended Strategies Essay

The article authors, Johnson and Kaplan looks at how management accounting has evolved over the years and within different industries and how those management accounting reports have failed to help mangers make decisions to reduce costs and improve productivity. The authors state that contemporary trends in competition, technology, and management demand major changes in the way organizations measure and manage costs and how they evaluate short- and long-term performance. The article takes a look at management accounting over varies periods of times and specific industries and discusses how at each period of time the management reports were used. For example, in the 19th century after the Industrial Revolution it was observed that gains could be earned by managing a hierarchical organization. The management system at the time focused on conversion costs and produced only summary results. Fast-forward a several years to roughly around 1925, we see that the management accounting practices that are practiced today had been developed by that time. They had been evolved to serve the control and informational needs of managers of increasingly complex and diverse organizations. As time progressed it is not until after the 1920s that the authors believe that evolution of management accounting did not keep the pace with the improvement in corporations’ product and process technologies. It is stated that the systems today provide misleading targets for managerial review. They fail to provide the relevant set of measures that reflect the technology, products, processes and competitive environments. Which has resulted in what they consider as today’s problems: distorted product costs, delayed and overly aggregated process control information, and short-term performance measures that do not reflect the increases or decreases in the organization’s economic position. Johnson and Kaplan conclude by stating that if companies fail to make modifications in their management accounting systems, their ability to be effective and efficient global competitors will be inhibited. Diagnostic Control Systems: Implementing Intended Strategies In chapter four, Robert Simons introduces what is known as the third lever of control: diagnostic control systems. These systems are defined as the backbone of traditional management control, and are designed to ensure predictable goal achievement. The other levers (Belief systems, Boundary Systems and Interactive Control Systems) are mentioned in the reading as well, however the focus of chapter four is to discuss the diagnostic control systems. He highlights three features that distinguish the control systems: (1) the ability to measure the outputs of a process, (2) the existence of predetermined standards against which actual results can be compared, and (3) the ability to correct deviations from standards. The chapter goes on to describe critical performance variables. Those variables as defined are those factors that must be achieved or implemented successfully for the intended strategy of the business to work. The term, â€Å"key success factors† can also be used. In which effectiveness and efficiency are the prime criteria for the selection measures used in diagnostic control systems to ensure that they are managed both effectively and efficiently. Kaplan and Norton uses the term â€Å"balanced scorecard† to describe a systematic way of analyzing critical performance variables and measures associated with intended strategies. This method allows managers to use measures from each of the four categories (Financial, Customer, Internal Business and Innovation & Learning Measures) simultaneously to guide their business toward the desired goals. The author conveys the message that equipping management systems to control strategy is not an easy task. Managers have to understand their strategies and be able to recognize the relationships between strategic and operating decisions and how they affect the bottom line.

Positioning View of McDonalds Competitive Advantage Essay

Positioning View of McDonalds Competitive Advantage - Essay Example According to the discussion McDonald’s emerges as a key player in the fast food industry and has worked relentlessly to attain a competitive advantage thereof.   The three major competitive strategies that a firm can follow include differentiation, focus, market segmentation, and low cost. In this sense, completive advantage can be conceived as the relative superiority in skills and resources. There are two major views of achieving competitive advantage, and they include resource-based view (RBV) and positioning view, which is construed as a consequence of RBV. In this discussion, the RBV and the portioning view will be defined with reference to the McDonald’s Company.This paper highlights that  resources and skills of a company are overly important as they are regarded as the major sources of competitive advantage. It is against this backdrop that the RBV is grounded and it proposes that in order to achieve competitive advantage, the assets and capabilities of a co mpany have presently scarce, not easily obtained in the market, non-substitutable and difficult to imitate besides furnishing economic value to the company. Whereas assets refer to the accumulated resource endowments of a company, capabilities are the skills that make it possible for the assets to be deployed in an advantageous manner. In regard to McDonald’s, the RBV of achieving competitive advantage is comprehended from the design and human resource dimension.  

Strategic Marketing Management of Wal-Mart Case Study

Strategic Marketing Management of Wal-Mart - Case Study Example As a result, it's British subsidiary ASDA, which already was proficient in the Britain, has made a successful business in U.K. Presently, the ASDA-Wal-Mart supercenters stand at the second level in the supermarket chain of U.K. It covers a huge market share of 17% in the U.K. After the takeover by Wal-Mart in 1999, the retail company has flourished by leaps and bounds. In 2004, ASDA operated a large number of stores in Scotland and employed a number of employees. The total number of store operated by ASDA in 2004 was 259 and the employee strength was 122000 (ASDA/WAL-MART: A Corporate Profile, 2004). The key to ASDA-Wal-Mart's colossal success is its strategic marketing management policies. The company has implemented some highly calculated and effective marketing strategies. It has always made an effort to render the best product to the customers in terms of quality and price. Simultaneously, it has imparted a feeling of associate partners to all its employees. Strategic marketing is a complex management technique for the identification of proper marketing opportunities. It aims to offer better values to the most profitable areas of the business without harming customer interests. The core marketing concepts are concerned with customer needs, customer-client relationship and supply of quality products at affordable prices. This includes a sophisticated market research that rests on customer feedback. A company is directed to build a competitive-edge for long-term activities with the application of various management techniques. The objective of strategic marketing management concerns with a wide array of activities. It involves an aggressive approach to capture the market share rapidly. While performing such kind of marketing management activity, a company needs to focus on the product diversity, the various geographical regions in which it is operating, the role of branding, the marketing channels it is using and the product quality it is offering. All such areas of marketing are required to improve for strategic marketing management. A giant retailer like Wal-Mart will need to re-focus all these issues to find the drawbacks and improve its service by correcting them (Strategic Marketing Management, 2009). The domain of strategic marketing management is vast and it includes internal as well as external analysis of the company. This report focuses only on the external analysis of the company. There are various models that have been put forward to express the external analysis of a company. Porter's five forces model is the most popular among them. The task of strategic analysis involves a high level of expertise and experience. The analysts must possess both the qualities to conduct a strategic analysis with ease. When an analyst uses Porter's five forces model, the planning process consists of a chain of steps. The

Corporate Governance and Compensation Research Paper

Corporate Governance and Compensation - Research Paper Example ance as â€Å"the system by which business corporations are directed and controlled†¦[specifying] the distribution of rights and responsibilities among different participants in the corporation such as the Board, managers, shareholders and other stakeholders, and spells out the rules and procedures for making decisions on corporate affairs† (OECD, 2004, 11). A third definition is articulated by economists: â€Å"Corporate governance is a field in economics that investigates how to secure/motivate efficient management of corporations by the use of mechanisms such as contracts, organizational designs and legislation† (Fernando, 2006, 13-15). More recent literature has expanded the coverage of corporate governance to include management and financial discipline, corporate social responsibility, business ethics, stakeholder participation in decision-making, and more recently, sustainable economic development in the country in which the business operates (16). Presented in the appendix is a figure showing the comparative descriptions of the OECD and UK’s Good Governance Standards for Public Service, as they measure up to the generic principles of good governance. Executive compensation as a concern of corporate governance Before the financial crisis, there was little reason for doubting the rising pay of chief executives as set by board. The general public perception is that the board acted in a regular and informed manner when they agreed in setting the compensation level of the CEO, and that corporate governance was implemented optimally from the top of the organization. The issues that arose out of the financial crisis, however, led to the belief that the process of determining CEO compensation is not as efficient as the public were led to believe, and that significant... This essay stresses that the case of Disney’s derivative suit involving Ovitz’s morally untenable compensation package highlights the huge discrepancy between what the law requires and what is required by good corporate governance, as far as executive compensation is concerned. Presently, in determining liability and propriety the law measures executive action by the minimum standard of due care and good faith, but this case shows that it is possible for a powerful CEO to meet the minimum and still cause severe pecuniary and organizational risk to the company and its shareholders. This paper makes a conclusion that the courts decided against the derivative suit filed by the shareholders, because the board of directors were deemed to have met the minimum standard of due care, and not to have acted in bad faith as defined by the law. This does not clear Disney executives, particularly its CEO and those directors who evidently connived with him, from meeting their moral and ethical responsibility towards their shareholders and stakeholders, and is an evident display of poor corporate governance. Executive compensation and the hiring of a successor must not be left to the whims and designs of a CEO who manipulates these matters in order to retain power for himself. The terms of executive compensation and succession should be considered as legitimate by social standards and the norms and cognitive impacts on the organization.

Human resources Management Assignment Example | Topics and Well Written Essays - 4250 words

Human resources Management - Assignment Example RasGas also supplies LNG at home and in addition to producing LNG for export markets in the amount of 37 million tonnnes each year, RasGas also supplies about 2.0 billion cubic feet of LNG at home. RasGas also ‘manages and operates’ the ‘Hellim 1 and Hellim 2 facilities’ which together produces 1.96 billion cubic feet of liquid hellim annually. RasGas is committed to increase production with a view to meeting energy demands both at home and abroad (RasGas, 2001-2013). †¦develop, produce and sell hydrocarbaons from the world’s largest non-associated gas field in a safe and environmentally responsible manner for the welfare of the State of Qatar and the satisfaction of our customers while maximizing shareholder value (RasGas, 2001-2013A). ..The safe and reliable production and delivery of products to a worldwide portfolio of customers and the superior execution of projects and technical services for our shareholders and stakeholders (RasGas, 2001-2013A). Thus, the overall strategy of RasGas as an organization is to provide high quantity, and high quality LNG in a manner that is environmentally friendly. RasGas also emphasizes efficiency and excellence in customer service, profit maximization for shareholders and satisfaction for a wider class of stakeholders. Sustainability is at the heart of RasGas’ strategy. According to RasGas, sustainability is the key to ‘business success’ (RasGas, 2001-2013B). Sustainability is therefore built on four pillars: performance improvement through competitiveness and innovation, improving relationships with stakeholders and shareholders, maintaining integrity through high operational and business standards and sustainable practices (see Figure 1) (RasGas, 2001-2013B). RasGas’ external labour market consists of operating a ‘fleet of 27 LNG carriers under long term charter agreements with ship owners’ (RasGas, 2001-2013). This fleet of carriers include ‘conventional, Q-Flex and Q-Max

Read Case study of A and others v The National Blood Authority and Essay

Read Case study of A and others v The National Blood Authority and others [2001] 3 All ER 289. Have cases since this decision de - Essay Example Indeed, the case of A and others v National Blood Authority and another became a landmark and certainly the first case in UK for being the first case in UK to succeed against the producer of a medical product. The Consumer Protection Act arising from this case certainly gained much footing and has since remained relatively unchanged with time. However, several other cases seem to enforce rather than change consumer law in UK. It is worth noting, however, that the case of A and others v National Blood Authority and another gained strong ground based on the fact that consumer protection was viewed from what the consumers are entitled to expect as opposed to the reasonable ability of the producer in delivering safe products. In the case of Worsley v Tambrands Ltd, Worsley argued that tampons manufactured by Tambrands were defective since the manufacturer did not provide clear warnings regarding the risk of toxic shock syndrome. However, this argument was rejected by the court based on t he argument that defectiveness of products is based on minimum standards (Howells and Weatherill 241-243). Therefore, the developments of Worsley v Tambrands Ltd’s case only affirmed the basis of consumer protection Act developed from A and others v National Blood Authority and another. One of the most recent cases BSS Group Plc v Makers (UK) Ltd (t/a Allied Services) [2011] seemed to bring a new twist to consumer protection Act. In this case, the important factor to be put into consideration is the obligation of the manufacturer to furnish the user with adequate information concerning the use and compatibility of a product (Bicknell web). However, this case seems to strengthen rather than change the provisions arising from A and others v National Blood Authority and another. The court ruling seemed to underpin the obligation of the manufacturer to provide adequate information on use of products. This had already been coined in the earlier case A and others v National Blood A uthority and another. Another, yet very recent case, Trebor Bassett Holdings Ltd & Anor v ADT Fire and security Plc [2011] also mirrored the already established consumer protection Act. According to the, the purchasers arguments CO2 fire suppression system they had purchased for their popcorn machinery in the factory was not fit for purpose. According to the case ruling, they failed to adequately notify the supplier of the intended use of the product they purchased and therefore, they could not have relied upon the supplier’s technical skills and reasonable judgment (Bailii web). However, a more interesting case Ide  v  ATB Sales Ltd (2007), provided a deeper mirror on consumer protection Act. From the case proceedings, the Judge posited that fatigue cracking was a probable cause of the fracture leading to the accident of the victim. However, proof of failure of the product during normal use had to be supplied adequately (Sweet and Maxwell web). The ruling in this case di d not actually change any aspect of consumer protection Act arising from A and others v National Blood Authority and another but to the contrary, only seemed to coin what had already been put forward. However, there seemed to be a

Assessments Essay Example | Topics and Well Written Essays - 1250 words

Assessments - Essay Example Science is a vast field, as it includes different experimental aspects of study area. Designed assessments of science subjects contribute to measure the growth and understanding of the subject. Assessments enhance students' learning and help teachers identify what students know and what they can do with their knowledge (IMS.Ode, 2011). This paper aims to design an assessment program for science students, which will include processes and instruments integral for students' learning. The designed assessment will be applicable for students of grade 6 for their science subject. Before designing an assessment program for students it is consider highly important to identify specific requirements of efficient activities that help insight students' knowledge of the subject (Allen, 2006). Likewise, at first place this assessment program will focus on reducing stress from both teachers and students and before practicing planned assessments, all assessment processes, practices and instruments wi ll be checked to note down their validity, reliability, and transparency (Pianta, 2012). Undoubtedly, paper pencil examination method is a traditional one and it is successful too, but it cannot completely evaluate students’ understanding and knowledge of the subject (Wang, 2011). Since, science is a vast subject and it is compulsory to design such an assessment that could ensure achievement of subject value, practice, research and communication outcomes. Thus, this assessment will include different activities and tasks to target each of these aforementioned goals in terms of learning outcomes. For learning and gaining expertise in any particular subject it is always important to develop interest in the subject. In the beginning of the session science teacher will ask students to write an informal essay on the topic, â€Å"Reasons for Selecting Science as a Major Subject in the Future†. This technique will help students use their imaginative and thinking power, because while writing essay they will get chance to recognize importance of scientific studies and scope of the subject in the professional career (Haines, 2004). Additionally, teachers will also get an idea about each student’s interest and dedication for the subject. This will also help teacher to design final assessment activities more effectively with respect to students’ interest and choices (Haines, 2004). Moreover, in the same class after finishing written essay students will be encouraged to come forward and read out summary of their respective essays. With this approach, it could be predicted that some students might hesitate to come forward and present in front of the entire class, but it will be teacher’s responsibility to provide equal opportunity to each student within available time and appreciated them for their future planning (Tobey, 2005). It will help develop and improve communication and presentation skills of the students. Subject teacher is going to monitor all students throughout the terms, and for this purpose, it could be suggested that after explaining a topic one or two times. Class discussion of that particular topic should be conducted to assess what students have understood from the teacher’s lectures and how well they are able to interpret their understanding about the topic (Race, 2005). As this assessment process targets science subject students, therefore teacher will be more emphasizing on examples, from routine life. Furthermore, for homework students will be given an interesting activity such as, they will have to paste or draw pictures relevant to the topic studied in the class. Although, it seems primary level activity, but it is most appropriate way of developing students’ interest in the subject and engage them in activities related to the subject (Race, 2005).

American law enforcement organizations Term Paper

American law enforcement organizations - Term Paper Example Objectives together constitute the main aim of this research. All objectives need to be studied in detail in order to fulfill the aim of this study and to answer the research question. Research design will elaborate on all the methods that have been used during the course of this research. Each objective requires a slightly different research approach and thus for each objective research methods have been used accordingly. Â   Â  There are two main methods of research, primary and secondary. Generally for the research purpose researchers’ start with the secondary data which has been already collected by someone else for some other research purpose. Such data can be used again and again for different research purposes. In this case, research has been conducted mainly using the secondary data. The sources of secondary data that have been used throughout the research include internet articles, reports, books, journals and the official websites of different law enforcement organ izations. Secondary data has an advantage over the primary data collection. Primary data is collected for the first time and may contain some loopholes. In the case of secondary data collection, data has already been collected by another entity and has been used for different research purposes. This provides with a satisfaction that the data available is authentic as it is used by various people for the research purpose. Another advantage of the secondary data is the time restraint. The time period for this research.... Local or state police, federal agencies, national police force, commerce bureaus, trade agencies and criminal justice organizations are some of the law enforcement agencies that form the hub of the state’s disciplinary activities. Law enforcement organizations are like departments and need to be well managed. Poor management of these departments results in poor law enforcement and inefficiency. The world is changing every day and is creating more and more challenges. Law enforcement agencies need to cope with these challenges by remaining up to date with the managerial procedural innovations so that they do not lack on their efficiency. The study focuses on these innovations and how they help in improving the efficiency of the law enforcement organizations. The study will not just discuss about the challenges faced by the law enforcement agencies, but will also ponder over the structural features of these organizations and what causes the law enforcement agencies to fail. The managerial aspect of these agencies will be the main focus. 2. Research Question Are federal law enforcement agencies better than the national law enforcement agencies when it comes to efficiency and meeting the 21st century challenges? 3. Aim and Objectives 2.1Aim The aim of this study is to find out the three most important ways through which the law enforcement organizations can improve on their efficiency. 2.2Objectives 1) The first and foremost objective of this research is to find out the main role of the law enforcement organizations in dealing with the daily affairs of any country. 2) The second objective is to study the challenges faced by these law enforcement organizations in the 21st Century and how do these law enforcement organizations

Modern variations of the Panopticon Essay Example | Topics and Well Written Essays - 1250 words

Modern variations of the Panopticon - Essay Example Bentham’s idea is that a panopticon, where the observed internalizes the presence of an unseen observer, enforces those rules via a psychological self-policing on the part of the observed. The panopticon prison, where prisoners are always within view of an observer hidden in a tower that sees all, is a very good representation of Bentham’s ideal panopticon. It is an examination of the ideas on the panopticon as it is applied in some paired relationships, exploring how checks and balances come into play in them, so that the two sides in effect are forced to play according to the rules. In effect, in this dyad relationship, the panopticon becomes a two-way mechanism, with implications that are particular to the parties in the pairing. This paper demonstrates this via a discussion of the Internet as a reciprocal modern panopticon. The internet fosters an arena where the observed follows the rules imposed on them by the law and by the authorities, and where the authorities, in their turn as ordinary users, are likewise compelled to follow the rules, and to make sure that the rules do not go too far in infringing on their personal rights under the law (Foucault 228). This first part of the essay talks about the writer's understanding of Foucault's Panopticon as a disciplinary power mechanism. The heart of the panopticon is the internalization of a power mechanism on the part of the observed, in a system designed in such a way that the observed knows that he is perennially being watched, and where the observer is forever hidden from view from the observed. This can be a paired process with both parties being observer and observed for each other. That is, the panopticon also a situation where two sides play mutual roles at the same time. In this sense it is reciprocal system that enforces the rules of the game via its very mechanism. The means of enforcement is the observed internalizing the rules, and acting in such a way as to enforce the rules, and to make the system self-perpetuating and stable. In a two-way system both have to play the role of the observed, and both internalize the rules (Foucault 228). In other words, the panopticon as a disciplinary power mechanism instills in the both parties the very rules of the game, and makes it in the best interest of both, in a way, to follow the rules without being told. What affects one affects the other in equal measure. The mechanism has within it the power to enforce. The knowledge that one is being watched at all times forces the observed to act in ways that do not violate the rules, or else face the consequences of the violation. It is self-perpetuating and becomes more effective the more it is internalized too. This is because both parties, as the observed, by internalizing the rules and the idea that he is being perpetually watched, go into themselves and look into all areas of their lives: thoughts, behavior - and seek compliance with the rules in all of those dimensions of his life. Both censor each other and are forced to follow the law as the observed, and as the observers enforcing the law (Foucault 226-228). In pairs where observation is reciprocal, the two parties balance each other's

Top 5 Essay Topics and Tips How to Be Original Essay Example for Free

Top 5 Essay Topics and Tips How to Be Original Essay The first writers on Earth were not actually writing – they were drawing pictures of the daily life. That was long before the actual writing started. Those days one did not have to be creative to â€Å"write† – a piece of chalk and a cave wall was more than enough. Luckily, we have evolved, and despite they write, the present Homo sapiens also try to deliver a unique, one-of-a-kind written message to the world. Believe it or not, creativity is very hard to achieve, and now you will find out why and what do with it. It seems like all the imaginable and unimaginable issues have been already discussed, described and published. So contributing a breath of fresh air to the endless stream of mass media looks impossible. Not only professional writers and journalists face the problem with being creative and original, but the students as well. Writing an essay, a term paper or a research paper can sometimes be compared to childbirth – with sweat and blood, a painful process that drives you crazy. That is why more and more students avoid all these troubles and turn to professional writing services for help. It is a great idea, though it will never make one original. Don’t be desperate – there are a few ways to keep your creativeness up. First of all – cheer up, buddy! It’s a simple psychological factor – the more negative thoughts you have in your head, the less creative and productive you are. Just smile, take your laptop/sheet of paper/notebook and start writing. Some instructors are likely to give you a topic. It makes your task much easier, as at least you know what to write about. But if you picked up a creative writing course – forget about it, most of the times you will be the one to come up with an interesting point to dwell on. At the stage of choosing the headline for your essay, the Internet or any published media can be of a great help. Try searching for the information you are interested in, what fascinates you most. This will keep up and feed your motivation, which is a vital thing for creativeness. Once you sifted through all the materials you found and came up with a topic you want to write on, you are ready to move further. Concentrate on your topic and write down your ideas – simply put your thoughts on the paper (thoughts like â€Å"I wanna eat† or â€Å"It was a great party last night† do not count!). Avoid going off-topic. Please note that in order to write something worthy and interesting, you are supposed and advised to spend at least 2-3 days on the writing process. Here I explain why: after you reviewed the articles on your chosen topic, your brain is overwhelmed with information, and to dot one’s is and cross one’s ts you need some rest. It’s better to make a research in the evening and start writing in the morning, with all the material in your hands. Fresh and creative ideas will fill your imagination for sure! If the very first stage has caused you troubles, you can use the following top 5 essay topics: Can love be associated with pain? Role models in your life Smoking – a bad habit or a disease? How to fight a depression? Pros and cons of having a pet I would like to sum up with a famous quote from Steve Jobs: â€Å"Stay hungry, stay foolish†. This phrase is the key to creativity. Dear reader, strive for knowledge, education and explore the world – nothing else will make you truly original.

Strong Brand Since The 1950s Marketing Essay

Strong Brand Since The 1950s Marketing Essay Brands are of high importance to companies since they have the ability to bring attributes and associations to consumers minds and add value to the producing company and the product itself (Kotler, 2003). Adidas is a globally very strong brand since the 1950s. Adidas manufactures products that cover a wide spectrum of the total global sport market. Their products are divided in three main categories which are: Adidas Sport Performance, Adidas Sport Heritage and Adidas Sport Style. The wide variety of products that Adidas manufactures is the core problem since it is difficult for the company to establish full communication with its potential consumers. As a result of adding value only to some of the product segments is the possible association, in the mind of the consumer, of the brand with only some of the segments. Adidas in the eyes of the consumers is mostly associated with football which as a sport is by far known as a mens sport. Women on the other hand the previous years werent prioritized for the communication and sales promotion strategies. But the last decades women are involved in sports in a much higher percentage equally to the one of men. This has led the female sportswear market and industry to grow rapidly in order to cover the gap. In many households women are making the final decisions for the products that will be bought. This has led many companies to change their advertising position and make their advertisements more attractive to women. Advertising and communication strategies are the power to create associations and perceptions to a brand and enhance its brand image so that the brand will be more attractive and interesting in the mind of the consumer (Nilson, Torsten, 2000). In order to succeed in reaching out to women Adidas followed the changes, adapted to the needs that modern women have and came up with a deal with world famous designer Stella McCartney developing a new product line extension known as Adidas by Stella McCartney. STRATEGIC DECISIONS Question 2 Prepare positioning maps and perceptual maps for the chosen-appointed subgroup. Present appropriate explanations in order to support your proposals. Positioning Map Male Innovative Classic Adidas Female Adidas by Stella McCartney is generally positioned in the positioning map as a new innovative stylish top collection designed for women. Sportswear apparel for women exists as a product line in Adidas product categories for many decades. The new top line collection by Stella McCartney is an innovative new collection in womens sportswear that combines performance and functionality with high quality. Perceptual Maps High School Graduate or more Some High School or less Economically strong Economically weak Adidas Adidas by Stella McCartney collection is mostly addressed to women who are educated and in a way wealthy because of their personal success in business environment as a result of personal achievements. By being economically strong women can buy this collection rather searching for products which are best value for money, or for special discounts and offerings; the Adidas by Stella McCartney does not have discounts or special offers. Big Spender Out of Fashion In Style Stingy Adidas Adidas by Stella McCartney new top line collection targets women who spend a lot of money in cloths and shoes and in sports apparel in general. They usually look to purchase sportswear by famous designers to fulfill their need and desire to be attractive, different and stand out in the crowd. Sport Active Expensive Cheap Sport Inactive Adidas The new top line collection Adidas by Stella McCartney is addressed to women who are sport active and are willing to pay more money to purchase sportswear designed by a famous designer. Big City Income 15000-30000 $ Income Rural Adidas Adidas by Stella McCartney is addressed to women living in big cities and having an average income of 23000$. In big cities the trends are a daily phenomenon and economically strong women want to be in the trend and always look stylish because of the competition that exists sometimes in big cities. Every successful woman likes to be different and attractive because of her stylish and famous designed outfit. ANALYSIS OF THE PROMOTIONAL MIX Question 3 Critically present the advertising and sales promotion objectives of the specific subgroup. Through advertising companies try to attach certain values to a product and make it more attractive to the needs of the consumer. Advertisements are paid announcements which usually target a specific market target group and are made or designed in a way to influence the consumer to buy the product or service being advertised. For that reason advertisements can be promoted by television, radio, newspapers, magazines and internet (Cohan, 2001). The last years Adidas has launched a new collection which is released every six months and is designed especially for women; it is called the Adidas by Stella McCartney. Adidas by Stella McCartney new collection is communicated and advertised through some types of media and the Adidas Group itself and both sides hope to increase the brand awareness and enhance the brand equity. Adidas believes that the most suitable way to communicate with women that their target group consists of is through womens magazines. For that reason every womans magazine in Greece has ads of the Adidas by Stella McCartney collection. These ads are published at the magazine issue that is around the date of the new collection release, before and during major holidays such as Christmas and Easter. The collections distribution on the other hand is limited. The head offices in Athens Greece control the places where the collection will be available for purchase. Such cities are the centre of Athens, Thessaloniki, and Larissa. But also in those cities only some selected stores have the permission to sell Stellas collection and that occurs after being very carefully chosen complying with a number of criteria. The image of Adidas and Stella McCartney can not be risked by having stock products. This new top product line is targeting people in large cities; on the other hand the Woman Indoor classic line is available in Adidas stores all over the world and is addressed to general women consumers. Because of the fact that Adidas is planning on spending a bigger budget every year on the woman segment the company looks for spokespersons that can promote their new line and add value to it. Of course Stella McCartney can be considered as the first but some other agreements have been made too. Such spokespersons who can also be considered are among other world famous athletes, tennis player Maria Kirilenko, and 400 meters hurdle runner Fani Halkia from Greece. Both of these athletes have signed not only to model but also to wear Adidas by Stella McCartney collection in every athletic event or major training day. Adidas does not prefer television ads to advertise the womans collection. The company uses the female magazines to organize events and festivals in large department stores and promote its womans collection through direct marketing communication. Also Adidas has never used discounts on Stella McCartneys collection. Instead the company prefers to organize fashion shows with runways, competitions and snack and beverage happenings to promote their top line collection for women. Question 4 Present the market segments that you propose to be targeted. The target group(s) shall be reported in demographic and mental-psychographic and purchase motives terms. Adidas main target group for their womens collection should be consisted of 20 and 35 year old women, maybe 37. Such a woman should live in a city or in a suburban area near a big city. She can still be in college or she is older, independent and working in preferred position in a company. The targeted segment is able and spends a large amount of money on clothes, shoes and apparel. In mental psychographic and purchase motive terms she can be characterized as sexy and seductive for the group of 20 to 27 and economically independent and successful for the group 28 to 35. She believes that by purchasing and having a world famous label in her cloths and shoes she improves her image, which is the reason why she buys world famous designers expensive brands for cloths and shoes, because by that purchase she believes that her well known cloths make her being perceived accordingly by her environment. That targeted woman is very stylish in a peculiar way sometimes but likes to capture attention and glances of the people around her, she likes giving stylish and purchase advises and generally she likes to be unique in a crowd. Sports on the other hand play a very important role in her everyday life. She wants to be perceived as an athlete for her leisure time but she takes the chance every time there is a competition. She wants her clothing to be functional but by all means they have to be stylish as well so that she can feel secure. She usually spends her time on training in different kinds of sports 3 to 4 times per week and simultaneously she can also have a subscription in a gym or even a fitness center for relaxation, because staying fit and being healthy is among her priorities. Question 5 In brief present your sales promotion (of any type) proposals, in order to support marketing communication program and help to succeed the 2010 sales goals of the specific Adidas subgroup. Since 2004 Adidas has decided to spend more and more economic resources every year to increase its brand awareness towards women. As result a business agreement, more like collaboration, was established with word famous designer Stella McCartney. The Adidas by Stella McCartney collection is more stylish and fashionable including each years new fashion and style trends and separating from the previous collections designed for women which were targeted more into sport type women. Having TV commercials is not suitable for the advertising of the new collection. TV advertisements cost a lot and are a difficult task to try communicating efficiently with the companys target group in all countries; plus if TV ads are not controlled effectively can lead to an overexposure of the collection and as a result to the negative perception of the brand. On the other hand special discounts and offerings are in a way forbidden because they could lead in negative perception of the brand. The new top line collection for women should increase the brand equity and enhance the companys and designers brand image. What Adidas can follow is a direct marketing strategy, which is already being followed. Adidas by Stella McCartney collection is currently advertised in the majority of the womens magazines in Greece. Also Adidas can benefit from the fact that its collection is offered in the countrys most known shopping centers and malls and with the collaboration of each magazine separately can organize fashion shows with models wearing and promoting that top line collection. Also happenings can be carried out in the shopping malls and departments with free snacks and drinks during the introduction of a new season collection. With such events Adidas promotes not only its brand and new collection but also the shopping store that distributes and sells its top line collection avoiding in that way overexposure. Another sales promotion activity which can be introduced in a more premium way is the organizing of special events especially for the women subscribers of the female and fashion magazines. Adidas can benefit from the magazines readers databases and invite them in a cocktail party for example with famous fashion models, artists and designers. In that way Adidas promotes its new collection, establishes better communication with women from its target group and enhances its brand image and brand equity.

Flow Cytometry for the Evaluation of Semen

Flow Cytometry for the Evaluation of Semen State of the Art in Sperm Assessment Using Flow Cytometry Abstract Flow cytometry is emerging as a substantial tool in the domain of modern andrology for the routine analysis of spermatozoa. Recent application of flow cytometry in the artificial insemination industry especially for pig is a new approach. Until very recent, analysis of semen samples was routinely performed by microscopical evaluation and manual techniques by laboratory operators; analysis is inclined due to comprehensive variability among observers, influencing its clinical validity. During last decade, to evaluate farm animal semen, variety of new flow cytometric techniques have been intercalated which made possible a wide spread evaluation of several sperm functionality and characteristics. Here in this paper, an initiative has been taken to explore numerous current flow cytometry developments pressing for andrological tests. After the invention of flow cytometry, sperm evaluation by traditional (microscopic) means became questioned and avoided due to the robust advantages of flow cytometry over the microscopic methods. By the recent development of diverse fluroscence probes, flow cytometry became capable of analyzing number of sperm characteristics like viability, capacitation, acrosomal integrity, membrane permeability, membrane integrity, mitochondrial status, DNA integrity, decondensation of DNA and differences between gametes based on sex. The application of flow cytometry to their detection allows increased numbers of spermatozoa to be assessed over a short time-period, provides the opportunity of working with small sample sizes, increases the repeatability of data obtained, removes the subjectivity of evaluation and allows simultaneous assessment of multiple fluorochromes. Thus, flow cytometry is a technique capable of generating significantly novel data and allows the design and execution of exper iments that are not yet possible with any other technique. Nowadays, semen evaluation using laboratory analyses is very meaningful to the artificial insemination industry to provide the most desired quality product to customers. Future development of flow cytometric techniques will permit further advances both in our knowledge and in the improvement of assisted reproduction techniques. In this paper, the main semen attributes that can be analyzed with fluorochromes and adapted for use with a flow cytometer will be reviewed and the relationship of these tests to fertility will be discussed. Introduction Up to now, semen evaluation is considered as the most important laboratory test that has enabled us to identify and predict clear-cut cases of fertility (Jarow et al., 2002), infertility or even of potential sub-fertility (Rodrà ­guez-Martà ­nez, 2007). Determination of the potential fertility of semen sample and, in the long run, of the male from which it has been collected is the ultimate goal of semen evaluations in clinically healthy sires. Now a days, many methods for the estimate the possible fertilizing capacity of a semen sample and, or in the word, of the male (reviewed by Dziuk 1996; Rodrà ­guez-Martà ­nez et al. 1997a; Rodrà ­guez-Martà ­nez and Larsson 1998; Saacke et al. 1998; Larsson and Rodrà ­guez-Martà ­nez 2000; Rodrà ­guez- Martà ­nez 2000, 2003; Popwell and Flowers 2004; Graham and Mocà © 2005; Gillan et al. 2005) are existing. The methods routinely accustomed for evaluation of the quality of a semen sample involved an evaluation of general appearance, volume, pH, sperm concentration, viability, morphology and motility. Most of these evaluations are based on microscopic analyses that only measure relatively a few numbers of spermatozoa within a population. In most of the cases, these are time-consuming; results obtained are controversial and are not translatable. It should also be noted that such conventional techniques are apt to extreme inter-ejaculate variation, even when the laboratory methodology has been standardized. In the wake of this information, new opportunities have arisen for the development of methods for the diagnosis of male infertility, many of which have been shown to exhibit a prognostic value that eludes conventional semen profiling. Moreover, ejaculated spermatozoa are nowadays handled for use in assisted reproductive technologies, such as the artificial insemination of chilled, frozen-thawed or sexed semen, and IVF. During this long processes, number of steps like semen extension, fluorophore loading, ultrav iolet and laser illumination, high-speed sorting, cooling and cryopreservation are followed, which create a scope to impose different degrees of change in sperm functionality followed by suffer of damage to sperm membranes, organelles or the DNA content. Therefore, although several assays have been developed to monitor these sperm parameters, recently it is being claimed by many groups that buck of those so-called procedures are incomplete, time consuming and laborious. Flow cytometry in diverse technical applications proposes many advantages for the analysis of sperm quality. Flow cytometry is a method where multiple fluorescences and light scattering can be induced allowing single cell or particles illumination in suspension while they flow very rapidly through a sensing area. The increasing use over the past decade of flow cytometry in the leading laboratories in human and veterinary andrology has dramatically increased our knowledge of sperm function under physiological and biotechnological conditions. Flow cytometers is capable to acquire data from several subpopulations within a sample in a few minutes, making it perfect for assessing heterogenous populations in a semen sample. Flow cytometry was initially developed in the 1960s, after that flow cytometry is performing automated separation of cells based on the unique recognition of cellular patterns in a population feasible (Hulett et al., 1969). Likewise, cellular patterns can be recognized by utilizing such a separation approach, in each cells within a population (Baumgarth and Roederer, 2000; Herzenberg et al., 2006). The first notion of flow cytometry development was for medical and clinical applications such as haematology and oncology. Although still much research is going on these medical areas and account for the vast majority of publications on this robust technique, but during the past few years it is being used in a diverse areas, such as bioprocess monitoring, pharmacology, toxicology, environmental sciences, bacteriology and virology. Together with elevated use in many areas, recent advancement of flow cytometry increased its application in the reproductive biology especially for andrology. Although flow cytometry may overestimate the population of unlabelled cells (Petrunkina and Harrison, 2009), plethora of research from our group in pig (Pena et al., 2003, 2004, 2005; Spjuth et al., 2007; Fernando et al., 2003; Saravia et al.,2005, 2007,2009; De Ambrogi et al., 2006; ) bull (Bergquist et al., 2007; Nagy et al., 2004; Januskauskas et al., 2003; Bergqvist et al., 2007; Hallap et al., 20 05, 2006;) stallion ( Kavak et al., 2003; Morrell et al., 2008) indicate that newly developed fluorescent stains and techniques of flow cytometry has made possible a more widespread analysis of semen quality at a biochemical, ultrastructural and functional level. Therefore, flow cytometry is the current technical solution for rapid, precisely reproducible assessment of sperm suspensions. In this review we have described potentiality and scope of flow cytometry for the evaluation of semen, and the way in which this technique can be used in clinical applications for andrology based on some of our previous experiences. Definition of flow cytometry The definition of a flow cytometer is ‘an instrument which measures the properties of cells in a flowing stream or ‘an instrument that can measure physical, as well as multi-colour fluorescence properties of cells flowing in a stream. In other word, cytometry is a method which measure physical and chemical attributes of cells or other particles. Such a measurement is made when cells or other particles pass in single file through some sort of measuring apparatus in a stream of fluid. The data obtained can be used to understand and monitor biological processes and develop new methods and strategies for cell detection and quantification. Compared to other traditional analytical tools, where a single value for each attribute is obtained for the whole population, flow cytometry provides data for each and every particle detected. As cells differ in their metabolic or physiological states, flow cytometry allows us not only to detect a particular cell type but also to find different subpopulations according to their structural or physiological parameters. Flow cytometry is a technique for measuring components (cells) and the properties of individual cells in liquid suspension. In essence, suspended cells are brought to a detector, one by one, by means of a flow channel. Fluidic devices under laminar flow define the trajectories and velocities that cells traverse across the detector, and fluorescence, absorbance, and light scattering are among the cell properties that can be detected. Flow sorting allows individual cells to be sorted on the basis of their measured properties, and one to three or more global properties of the cell can be measured. Flow cytometers and cell sorters make use of one or more excitation sources and one or two fluorescent dyes to measure and characterize several thousands of cells per second. Flow cytometry presents objective and precise results (Bunthof et al., 2001; Shleeva et al., 2002), which help to overcome the problems with the manual methods described above. Function and types of flow cytometry A flow cytometer is made of three main systems, fluidics, optics and electronics. ItI It can acquire data on all subpopulations within a sample, making it ideal for assessment of heterogenous population, such as spermatozoa. The adaptation of flow cytometry to sperm assessment came in to function when it was used for measuring their DNA content (Evenson et al., 1980) and its application for analyzing semen has been increased rapidly in last decade. Flow cytometry is now applied for the evaluation semen such as sperm viability, acrosomal integrity, mitochondrial function, capacitation status, membrane fluidity, DNA status and so on. Continuous innovation of new fluorescent stains and techniques facilitated the flow cytometric evaluation of spermatozoa. Flow cytometry allows the observation of physical characteristics, such as cell size, shape and internal complexity, and any component or function of the spermatozoon that can be detected by a fluorochrome or fluorescently labeled compound. The analysis is objective, has a high level of experimental repeatability and has the advantage of being able to work with small sample sizes. Flow cytometry also has the capacity to detect labeling by multiple fluorochromes associated with individual spermatozoa, meaning that more than one sperm attribute can be assessed simultaneously. This feature has an added benefit for semen analysis, as few single sperm parameters show significant correlation with fertility in vivo for semen within the acceptable range of normality (Larsson and Rodriguez-Martinez, 2000) and it is the general statistics that the more sperm parameters can be tested, the more accurate the fertility prediction becomes (Amman and Hammerstedt, 1993). There are two main types of flow cytometers-analysers and sorters are in use. Together with data collection on cells, sorters have the potentiality to sort cells with particular properties (defined by the flow cytometer operator) to extremely high purities. There are also a number of commercial flow cytometers that have been developed for particular analytical requirements. Partec manufacture a Ploidy Analyser and also a Cell Counter Analyser. Optoflow has developed a flow cytometer for the rapid detection, characterization and enumeration of microorganisms. Luminex is developing technology for multiplexed analyte quantitation using a combination of microspheres, flow cytometry and high speed digital processing. Advantages of FC compared to other conventional techniques to explore sperm structure and function Use of authentic assays in the fertility clinic and artificial insemination industries increasing day by day. In this respect, use of flow cytometry might be an important attempt to resolve sustaining problem with so called commonly used manual method for the semen analysis. An additional source of laboratory variation is the low number of sperms analyzed with such techniques. It is worth mentinign here that so called method deal only with few hundred sperm. When we deal with such a few sperm population, there is a possibility that obtained result might not be statistically significant (Russel and Curtis, 1993). The methods which are frequently used are enable to determine sperm concentration (Jorgensen et al., 1997), motility or morphology only (Keel et al., 2002). Objectivity, cell number measured, speed of count and precision are the advantages of flow cytometry to conventional light microscopy techniques (Spano and Evenson, 1993). The technique now a days has been used to determi ne a number of factors including those of acrosome status, membrane integrity, mitochondrial function as well as multiparameter measurement in human (Garrido et al., 2002). Flow cytometry has the ability to analyze thousands of cells in few minutes. In our series of studies, we demonstrated the feasibility and reproducibility of an automated method to evaluate sperm cell type, count, and viability in human boar samples. In our hand, the precision of the flow cytometric analysis is satisfactory in a diverse species (boar, bull, stallion etc), and the observed errors were significantly better than those obtained from the so-called manual methods. Although there are diverse benefits of flow cytometer for the analysis of semen, feasibility of applying flow cytometry sometimes restricted to researcher due to the high outlay and difficulties of operation associated with the requirement of a skilled operator. Further, a flow cytometer is very large and cannot resist shocks associated with movement, and it also requires much space in the laboratory. Whatever may be the limitation, the development of more affordable ‘‘bench-top flow cytometers in recent time raised the potential essentialities to semen analysis. If the further application of flow cytometric analysis is considered further, it might be seen that it is growing popularities as a technique for assessing more than one sperm attribute, simultaneously. Compared to traditional microscopic techniques, flow cytometry analysis is allowing to give a far more simplified and objective method of semen analysis, especially in relation to fertilization with acrosome reactivity potential of spermatozoa (Uhler et al., 1993; Purvis et al., 1990; Carver-Ward et al., 1996). A large number of different techniques to estimate sperm concentration have been reported. In the mid-1990s a series of fixed-depth disposable slides were evaluated as rapid and effective pieces of equipment for the estimate of sperm concentration. Data from a number of preliminary studies proposed that, at least in the 20-mm-depth format, such chambers resulted in a noticeable underestimate of sperm concentration compared to the gold standard (improved Neubauer hemocytometer). According to the World Health Organization that ‘‘such chambers, whilst convenient in that they can be used without dilution of the specimen, might lead to inaccuracy (World Health Organization, 1999). Data from Tomlinson and colleagues indicate that two proprietary disposable slides (Microcell, Conception Technologies, San Diego, Calif; Leja, Leja Products, BV Nieuw- Vennep, The Netherlands) can result in a lower concentrations of sperm compared to the hemocytometer method (Tomlinson et al., 2001) . In contrast, plenty of reports document unacceptable differences between different laboratories and even between different individuals, although fewer studies attempt to address these issues. So, what is wrong? Improvement of semen quality testing has been emphasizing in some reports (Jorgensen et al., 1997; Keel et al., 2000). But due to low number of sperm evaluation by the conventional method results in poor reproducibility. These problems might be overcome when using flow cytometry. The validation of method is a challenge due to its essentiality of having specific, precise, objective, and accurate evaluation to establish a correlation of fertility data or to predict potential of a semen sample accurately (Amann, 1989). In a fertility clinic, precision of data in important as the result of semen analysis is frequently used to manage fertility of a patient and treatment of the unfertile couples. Thus, it is important to take into consideration within and between laboratory variations for successful infertility treatments. Sometimes its a matter of argument that compared to flow cytometry, fluorescent microscopy evaluate â€Å"patterns of fluorescence rather than the fluorescence intensity. Flow cytometer has the lack of ability to discriminate sperm containing a fluorescent marker bound to the equatorial segment or over one of the acrosomal membranes (Parinaud et al., 1993; Mortimer and Camenzind, 1989; Mortimer et al., 1987). Tao et al. (1993) compared flow cytometry and epifluorescent microscopy with various lectins and indicated that there is almost no difference between methodologies for detection of the acrosome reaction. However, it has been argued that lectins do not bind specifically to the acrosomal region of the sperm (Purvis et al., 1990; Holden and Trounson, 1991) and that other binding sites can be easily distinguished by epifluorescence microscopy, whereas flow cytometry identifies the signal from the entire sperm. Additionally, conventional light microscopic semen assessment is increasingly being replaced by fluorescent staining techniques, computer-assisted sperm analysis (CASA) systems, and flow cytometry (PenËÅ"a et al., 2001; Verstegen et al., 2002). Additional advantages over existing techniques are that this approach is faster than the hemacytometer and that cellular debris, fat droplets, and other particulate material in extended semen are not erroneously counted as sperm, as often occurs with electronic cell counters. This method can also be used to determine the number of somatic cells in a semen sample. Application of flow cytometry for sperm count Sperm count is an important predominant factor for the evaluation of sperm fertility potential. Accurate determination of sperm cell concentration is critical especially in AI industry because it provides assurance to customers that straws of extended semen contain the sperm numbers indicated which will help to decide appropriate doze especially for pig. Accuracy of sperm count is a common problem in the andrological laboratories and accurate measure of sperm concentration is particularly important for export in which verification of numbers may be required. Routine sperm counts can help to identify possible processing errors within a specific batch of semen or on a particular day, should those errors occur. As sperm counting procedures become more refined, routine counting can be used to monitor subtle changes in daily semen processing that might affect the number of sperm packaged in a straw. Every time new and more accurate methods for the sperm count determinations are coming and being replaced by the older ones. Some laboratories are trying the Maklerm counting chamber (Se if- Medical, Haifa, Israel) and other improved hemacytometers, such as the MicroCellTM (Fertility Technologies, Inc., Natick, MA); however, these techniques will likely have standard lems similar to those associated with the standard hemacytometers. Although hemacytometers are routinely used for sperm counts, due to the slow process and need for multiple measurements of each sample, the chance of error increase. Freund and Carol (ref) stated that a difference of 20% were not unusual between the determinations by the same technician. Electronic counters provide much more rapid counting, are easier to use, and give more repeatable results among technicians. However, those instruments tend to include in the sperm count any somatic cells present, immature sperm forms, cytoplasmic droplets, debris, and bacteria, thereby inflating the concentration value (Ref). Spectrophotometer is recently being used in the AI industries to assess sperm concentration by determining turbidity of a semen s ample using an instrument previously calibrated for sperm concentration with a hemacytometer or Coulter counter (Ref). The accuracy of this method depends on the methods used for spectrophotometer calibration. Although, sperm concentration can also be determined by spectrophotometrically, the debris present in the raw semen crease problem with misestimation. Sperm number in the frozen thawed semen is difficult to ascertain as most of the extender contain egg yolk particles, fats and other particles which affect measurement of sperm with electric cell counter or spectrophotometers (Evenson et al., 1993). On the other hand flow cytometry created possibilities of a rapid determination of sperm number in a precise form. It is the flow cytometry which can reduce intra-laboratory and inter-laboratory variation and conflict regarding sperm concentration assessment. Computer assisted semen analyzer is robust technique for analyzing sperm movement which can count sperm as well; but such an a nalyzer most of the cases use some counting chamber or hemacytometer which itself can generate error. Although, hemacytometer was originally developed for blood cell counting, its use is now diverse including andrological laboratories for sperm counting. Around two-decade ago flow cytometry was suggested for sperm numbers in straws of cryopreserved bull semen. Christensen et al. (-) observed similar results for sperm count with flow cytometry and hemocytometer for a number of species. Now a day a simultaneous determination of sperm viability and sperm concentration is possible which can avoid the chance of occurring differences between ejaculates leading lack of coordination with field fertility and laboratory analyses. Thus the present technology is more precise which can get rid of variation from handling the sperm sample and variation from pipetting and the analysis itself. Further, Prathalingam et al. (2006) concluded that there is similarities for sperm count result between flow cytometry and two newly approached method (image analysis and fluorescent plate reader) for sperm counting. Though, use of fluorescent plate was emphasized due to low cost and allowing large number of cells counting from a large number of ejaculates. Although flow cytometry has become a valuable instrument for andrological determinations, it is also blamed that sperm concentration by flow cytometry signify a higher value than the real one. The possibility arise might be due to that semen samples often contain some alien materials such as immature germ cells, epithelial cells, blood cells, cytoplasmic droplet, cellular debris etc. In the same way, frozen semen has higher chance to introduce such material as they contain diluents components especially egg yolk particles. These particles and cell debris might have frontal and side light scatter parameters those are similar to spermatozoa. Such sperm-count-overestimation problem arisen in our cases also, especially when we deal with frozen semen. Further it is also claimed that flow cytometry has a tendency to overestimate viable spermatozoa. We are also experienced with such trouble which we guess might be due to that egg particles of extender are considered as viable cell as for it s staining pattern. Our yet to publish data indicate that this problem can be mimic by a centrifugation process and by using low concentration sample for evaluation with flow cytometry. Very recently Petrunkina and Harrison (2009) proposed a mathematical equation for fixing this flow cytometric sperm counting. Thus much research is going on and we hope such discrepancy will completely be resolved near future to get advantage from this robust technology for sperm counting. Flow cytometry for detecting sperm intactness -Viability of spermatozoa The viability of spermatozoa is a key determinant of sperm quality and prerequisite for successful fertilization. Viability of spermatozoa can be assessed by numerous methods, but many are slow and poorly repeatable and subjectively assess only 100 to 200 spermatozoa per ejaculate. Merkies et al. (2000) compared different methods of viability evaluation. They concluded that Eosin-nigrosin overestimate viability while fluorescent microscope and flow cytometry estimate similar trend of viability. Current flow cytometric procedures are able to simultaneously evaluate sperm cell viability together with some other attributes. This method has been successfully used for assessing spermatozoa viability in men (Garner and Johnson, 1995), bulls (Garner et al., 1994; Thomas et al., 1998), boars (Rodrà ­guez-Martà ­nez, 2007; Garner and Johnson, 1995; Garner et al., 1996), rams (Garner and Johnson, 1995), rabbits (Garner and Johnson, 1995), mice (Garner and Johnson, 1995; Songsasen et al., 199 7), poultry and wildfowl (Donoghue et al., 1995; Blanco et al., 2000) and honey bees (Collins and Donoghue, 1999; Collins, 2000) and in fish (Martin Flajshans et al., 2004). Considerable information has accumulated on the use of fluorescent staining protocols for assessing sperm viability (Evenson et al., 1982). The SYBR 14 staining of nucleic acids, especially in the sperm head, was very bright in living sperm. Good agreement was observed between the fluorescent staining method and the standard eosin-nigrosine viability test; the flow cytometric method showed a precision level higher than that of the manual method. One of the first attempts to assess sperm viability utilized rhodamine 123 for determining potentiality of mitochondrial membrane while ethidium bromide for membrane integrity through flow cytometry (Garner et al., 1986). Other combinations that have been used to examine the functional capacity of sperm are carboxyfluorescein diacetate (CFDA) and propidium iodide (PI) (Garner et al., 1988; Watson et al., 1992); carboxydimethylfluorescein diacetate (CMFDA), R123, and PI (Ericsson et al., 1993; Thomas and Garner, 1994); and PI, pisum sativum agglutinin (PSA), and R123 (Graham et al., 1990). The most generally used sperm viability stain combinations is SYBR-14 and PI at present. This stains are now sold commercially as live/dead viability kit. When these two stains are combinely used, the nuclei of viable sperm take fluoresce green and membrane integrity lost cells take red stain. This staining technique has been used in a number of species, including the boar (Garner and Johnson, 1995; Saravia et al.,2005, 2007,2009). Although species differences do exist in the function of spermatozoa, the Live/Dead stain may similarly have no adverse affect on fertilization in the equine, although it remains to be tested in this species. Recently a new instrument (Nucelocounter-SP100) has been introduced to evaluate sperm concentration [11] and viability. Due to the small size and low cost, this instrument has been attracted for field measurements of both concentration and viability. In our hand this instrument was also became useful for the quick measurement of sperm concentration an d viability in stallion (Morrell et al., 2010). Fluorescent probes such as H33258, requiring flow cytometric analysis with a laser that operates in the ultraviolet light range, are less commonly used as this is not a standard feature on the smaller analytical machines. However, one alternative is to use a fluorometer. A fluorometer is a relatively low-cost piece of portable equipment that permits a rapid analysis to be carried out on a sample. Januskauskas et al. (2001) used H33258 to detect nonviable bull spermatozoa by fluorometry and obtained an inverse correlation between the damaged cells per cent and the field fertility. Another option is fluorescent attachments for computer-assisted semen analysis devices. For example, the IDENT fluorescence feature of the Hamilton-Thorne IVOS permits staining with H33258 allowing an assessment of sperm viability to be made along with motility. Fluorochromes used to assess sperm viability by both approach could be utilized in combination with each other. In that case, when CFDA is used combined with PI, three populations of cells as live, which are green; dead, which are red; and a third population which is stained with both and represents dying spermatozoa can be identified. This combination was found useful by Almlid and Johnson (1988) for frozen-thawed boar spermatozoa for monitoring membrane damage at the time of evaluation of various freezing protocols. Further, Harrison and Vickers (1990) also noticed that this combination with a fluorescent microscope is effective indicator of viability of fresh, incubated or cold-shocked spermatozoa in boar and ram. Contrasting to these, Garner et al. (1986) was failed to find a relationship between bull sperm viability and fertility when using combination of CFDA/PI . Flow cytometry for evaluating sperm viability appears to be a precious tool in the AI industry. When a high number of sperm is packed in each insemination dose, the effect of selecting the best ejaculates according to sperm viability has a relatively limited effect. However, sperm viability might be more important when combined with low-dose inseminations. The FACSCount AF flow cytometer also determines sperm concentration accurately and precisely during the same analysis (Christensen et al., 2004a). The combined assessment of sperm viability and concentration appears to be useful in the wake of improving quality control at AI stations. Because of the results of this trial, this method has been implemented by Danish AI stations (Christensen et al., 2005). Relatively bright fluorescence was found also in the mitochondrial sheath of living sperm. But the mechanism and mode of action by which SYBR-14 binds to the DNA of sperm is not known. It is know that PI stains nucleic acids by inte rcalating between the base pairs (Krishan, 1975). Viability stains can also be used in conjugation with fluorescently labeled plant lectins for simultaneous assessment of the plasma membrane integrity and the acrosome integrity (Nagy et al., 2003). It is conceivable that assessment of viability using SYBR-14 dye does not damage spermatozoa, since Garner et al. (5) found that insemination of boar sperm stained with SYBR-14 did not compromise fertilization or even the development of flushed porcine embryos in vitro. Non-viable sperms can be detected using the membrane-impermeable nucleic acid stains which positively identify dead spermatozoa by penetrating cells with damaged membranes. Plasma membrane which is intact will not permit these stains entering into the spermatozoa and staining the nucleus. Most frequently used stains include phenanthridines, for example propidium iodide (PI; (Matyus, 1984) ethidium homodimer-1 (EthD-1; (Althouse et al., 1995), the cyanine Yo-Pro (Kavak, 2003) and the bizbenzimidazole Hoechst 33258 (Gundersen and Shapiro, 1984). After a series of comparison between fertility of cryopreserved stallion spermatozoa with a number of laboratory assessments of semen quality as assessed by flow cytometry using PI, Wilhelm et al. (1996) concluded that viability is the single laboratory assay that correlated with fertility. -Sperm plasma membrane integrity Although the sperm plasma membrane covers the entire cell, it consists of three distinct membrane compartments, one which covers the outer acrosomal membrane, one which covers the post acrosomal portion of the sperm head, and one which covers the middle and principal pieces. Sperm membrane is directly or ind Flow Cytometry for the Evaluation of Semen Flow Cytometry for the Evaluation of Semen State of the Art in Sperm Assessment Using Flow Cytometry Abstract Flow cytometry is emerging as a substantial tool in the domain of modern andrology for the routine analysis of spermatozoa. Recent application of flow cytometry in the artificial insemination industry especially for pig is a new approach. Until very recent, analysis of semen samples was routinely performed by microscopical evaluation and manual techniques by laboratory operators; analysis is inclined due to comprehensive variability among observers, influencing its clinical validity. During last decade, to evaluate farm animal semen, variety of new flow cytometric techniques have been intercalated which made possible a wide spread evaluation of several sperm functionality and characteristics. Here in this paper, an initiative has been taken to explore numerous current flow cytometry developments pressing for andrological tests. After the invention of flow cytometry, sperm evaluation by traditional (microscopic) means became questioned and avoided due to the robust advantages of flow cytometry over the microscopic methods. By the recent development of diverse fluroscence probes, flow cytometry became capable of analyzing number of sperm characteristics like viability, capacitation, acrosomal integrity, membrane permeability, membrane integrity, mitochondrial status, DNA integrity, decondensation of DNA and differences between gametes based on sex. The application of flow cytometry to their detection allows increased numbers of spermatozoa to be assessed over a short time-period, provides the opportunity of working with small sample sizes, increases the repeatability of data obtained, removes the subjectivity of evaluation and allows simultaneous assessment of multiple fluorochromes. Thus, flow cytometry is a technique capable of generating significantly novel data and allows the design and execution of exper iments that are not yet possible with any other technique. Nowadays, semen evaluation using laboratory analyses is very meaningful to the artificial insemination industry to provide the most desired quality product to customers. Future development of flow cytometric techniques will permit further advances both in our knowledge and in the improvement of assisted reproduction techniques. In this paper, the main semen attributes that can be analyzed with fluorochromes and adapted for use with a flow cytometer will be reviewed and the relationship of these tests to fertility will be discussed. Introduction Up to now, semen evaluation is considered as the most important laboratory test that has enabled us to identify and predict clear-cut cases of fertility (Jarow et al., 2002), infertility or even of potential sub-fertility (Rodrà ­guez-Martà ­nez, 2007). Determination of the potential fertility of semen sample and, in the long run, of the male from which it has been collected is the ultimate goal of semen evaluations in clinically healthy sires. Now a days, many methods for the estimate the possible fertilizing capacity of a semen sample and, or in the word, of the male (reviewed by Dziuk 1996; Rodrà ­guez-Martà ­nez et al. 1997a; Rodrà ­guez-Martà ­nez and Larsson 1998; Saacke et al. 1998; Larsson and Rodrà ­guez-Martà ­nez 2000; Rodrà ­guez- Martà ­nez 2000, 2003; Popwell and Flowers 2004; Graham and Mocà © 2005; Gillan et al. 2005) are existing. The methods routinely accustomed for evaluation of the quality of a semen sample involved an evaluation of general appearance, volume, pH, sperm concentration, viability, morphology and motility. Most of these evaluations are based on microscopic analyses that only measure relatively a few numbers of spermatozoa within a population. In most of the cases, these are time-consuming; results obtained are controversial and are not translatable. It should also be noted that such conventional techniques are apt to extreme inter-ejaculate variation, even when the laboratory methodology has been standardized. In the wake of this information, new opportunities have arisen for the development of methods for the diagnosis of male infertility, many of which have been shown to exhibit a prognostic value that eludes conventional semen profiling. Moreover, ejaculated spermatozoa are nowadays handled for use in assisted reproductive technologies, such as the artificial insemination of chilled, frozen-thawed or sexed semen, and IVF. During this long processes, number of steps like semen extension, fluorophore loading, ultrav iolet and laser illumination, high-speed sorting, cooling and cryopreservation are followed, which create a scope to impose different degrees of change in sperm functionality followed by suffer of damage to sperm membranes, organelles or the DNA content. Therefore, although several assays have been developed to monitor these sperm parameters, recently it is being claimed by many groups that buck of those so-called procedures are incomplete, time consuming and laborious. Flow cytometry in diverse technical applications proposes many advantages for the analysis of sperm quality. Flow cytometry is a method where multiple fluorescences and light scattering can be induced allowing single cell or particles illumination in suspension while they flow very rapidly through a sensing area. The increasing use over the past decade of flow cytometry in the leading laboratories in human and veterinary andrology has dramatically increased our knowledge of sperm function under physiological and biotechnological conditions. Flow cytometers is capable to acquire data from several subpopulations within a sample in a few minutes, making it perfect for assessing heterogenous populations in a semen sample. Flow cytometry was initially developed in the 1960s, after that flow cytometry is performing automated separation of cells based on the unique recognition of cellular patterns in a population feasible (Hulett et al., 1969). Likewise, cellular patterns can be recognized by utilizing such a separation approach, in each cells within a population (Baumgarth and Roederer, 2000; Herzenberg et al., 2006). The first notion of flow cytometry development was for medical and clinical applications such as haematology and oncology. Although still much research is going on these medical areas and account for the vast majority of publications on this robust technique, but during the past few years it is being used in a diverse areas, such as bioprocess monitoring, pharmacology, toxicology, environmental sciences, bacteriology and virology. Together with elevated use in many areas, recent advancement of flow cytometry increased its application in the reproductive biology especially for andrology. Although flow cytometry may overestimate the population of unlabelled cells (Petrunkina and Harrison, 2009), plethora of research from our group in pig (Pena et al., 2003, 2004, 2005; Spjuth et al., 2007; Fernando et al., 2003; Saravia et al.,2005, 2007,2009; De Ambrogi et al., 2006; ) bull (Bergquist et al., 2007; Nagy et al., 2004; Januskauskas et al., 2003; Bergqvist et al., 2007; Hallap et al., 20 05, 2006;) stallion ( Kavak et al., 2003; Morrell et al., 2008) indicate that newly developed fluorescent stains and techniques of flow cytometry has made possible a more widespread analysis of semen quality at a biochemical, ultrastructural and functional level. Therefore, flow cytometry is the current technical solution for rapid, precisely reproducible assessment of sperm suspensions. In this review we have described potentiality and scope of flow cytometry for the evaluation of semen, and the way in which this technique can be used in clinical applications for andrology based on some of our previous experiences. Definition of flow cytometry The definition of a flow cytometer is ‘an instrument which measures the properties of cells in a flowing stream or ‘an instrument that can measure physical, as well as multi-colour fluorescence properties of cells flowing in a stream. In other word, cytometry is a method which measure physical and chemical attributes of cells or other particles. Such a measurement is made when cells or other particles pass in single file through some sort of measuring apparatus in a stream of fluid. The data obtained can be used to understand and monitor biological processes and develop new methods and strategies for cell detection and quantification. Compared to other traditional analytical tools, where a single value for each attribute is obtained for the whole population, flow cytometry provides data for each and every particle detected. As cells differ in their metabolic or physiological states, flow cytometry allows us not only to detect a particular cell type but also to find different subpopulations according to their structural or physiological parameters. Flow cytometry is a technique for measuring components (cells) and the properties of individual cells in liquid suspension. In essence, suspended cells are brought to a detector, one by one, by means of a flow channel. Fluidic devices under laminar flow define the trajectories and velocities that cells traverse across the detector, and fluorescence, absorbance, and light scattering are among the cell properties that can be detected. Flow sorting allows individual cells to be sorted on the basis of their measured properties, and one to three or more global properties of the cell can be measured. Flow cytometers and cell sorters make use of one or more excitation sources and one or two fluorescent dyes to measure and characterize several thousands of cells per second. Flow cytometry presents objective and precise results (Bunthof et al., 2001; Shleeva et al., 2002), which help to overcome the problems with the manual methods described above. Function and types of flow cytometry A flow cytometer is made of three main systems, fluidics, optics and electronics. ItI It can acquire data on all subpopulations within a sample, making it ideal for assessment of heterogenous population, such as spermatozoa. The adaptation of flow cytometry to sperm assessment came in to function when it was used for measuring their DNA content (Evenson et al., 1980) and its application for analyzing semen has been increased rapidly in last decade. Flow cytometry is now applied for the evaluation semen such as sperm viability, acrosomal integrity, mitochondrial function, capacitation status, membrane fluidity, DNA status and so on. Continuous innovation of new fluorescent stains and techniques facilitated the flow cytometric evaluation of spermatozoa. Flow cytometry allows the observation of physical characteristics, such as cell size, shape and internal complexity, and any component or function of the spermatozoon that can be detected by a fluorochrome or fluorescently labeled compound. The analysis is objective, has a high level of experimental repeatability and has the advantage of being able to work with small sample sizes. Flow cytometry also has the capacity to detect labeling by multiple fluorochromes associated with individual spermatozoa, meaning that more than one sperm attribute can be assessed simultaneously. This feature has an added benefit for semen analysis, as few single sperm parameters show significant correlation with fertility in vivo for semen within the acceptable range of normality (Larsson and Rodriguez-Martinez, 2000) and it is the general statistics that the more sperm parameters can be tested, the more accurate the fertility prediction becomes (Amman and Hammerstedt, 1993). There are two main types of flow cytometers-analysers and sorters are in use. Together with data collection on cells, sorters have the potentiality to sort cells with particular properties (defined by the flow cytometer operator) to extremely high purities. There are also a number of commercial flow cytometers that have been developed for particular analytical requirements. Partec manufacture a Ploidy Analyser and also a Cell Counter Analyser. Optoflow has developed a flow cytometer for the rapid detection, characterization and enumeration of microorganisms. Luminex is developing technology for multiplexed analyte quantitation using a combination of microspheres, flow cytometry and high speed digital processing. Advantages of FC compared to other conventional techniques to explore sperm structure and function Use of authentic assays in the fertility clinic and artificial insemination industries increasing day by day. In this respect, use of flow cytometry might be an important attempt to resolve sustaining problem with so called commonly used manual method for the semen analysis. An additional source of laboratory variation is the low number of sperms analyzed with such techniques. It is worth mentinign here that so called method deal only with few hundred sperm. When we deal with such a few sperm population, there is a possibility that obtained result might not be statistically significant (Russel and Curtis, 1993). The methods which are frequently used are enable to determine sperm concentration (Jorgensen et al., 1997), motility or morphology only (Keel et al., 2002). Objectivity, cell number measured, speed of count and precision are the advantages of flow cytometry to conventional light microscopy techniques (Spano and Evenson, 1993). The technique now a days has been used to determi ne a number of factors including those of acrosome status, membrane integrity, mitochondrial function as well as multiparameter measurement in human (Garrido et al., 2002). Flow cytometry has the ability to analyze thousands of cells in few minutes. In our series of studies, we demonstrated the feasibility and reproducibility of an automated method to evaluate sperm cell type, count, and viability in human boar samples. In our hand, the precision of the flow cytometric analysis is satisfactory in a diverse species (boar, bull, stallion etc), and the observed errors were significantly better than those obtained from the so-called manual methods. Although there are diverse benefits of flow cytometer for the analysis of semen, feasibility of applying flow cytometry sometimes restricted to researcher due to the high outlay and difficulties of operation associated with the requirement of a skilled operator. Further, a flow cytometer is very large and cannot resist shocks associated with movement, and it also requires much space in the laboratory. Whatever may be the limitation, the development of more affordable ‘‘bench-top flow cytometers in recent time raised the potential essentialities to semen analysis. If the further application of flow cytometric analysis is considered further, it might be seen that it is growing popularities as a technique for assessing more than one sperm attribute, simultaneously. Compared to traditional microscopic techniques, flow cytometry analysis is allowing to give a far more simplified and objective method of semen analysis, especially in relation to fertilization with acrosome reactivity potential of spermatozoa (Uhler et al., 1993; Purvis et al., 1990; Carver-Ward et al., 1996). A large number of different techniques to estimate sperm concentration have been reported. In the mid-1990s a series of fixed-depth disposable slides were evaluated as rapid and effective pieces of equipment for the estimate of sperm concentration. Data from a number of preliminary studies proposed that, at least in the 20-mm-depth format, such chambers resulted in a noticeable underestimate of sperm concentration compared to the gold standard (improved Neubauer hemocytometer). According to the World Health Organization that ‘‘such chambers, whilst convenient in that they can be used without dilution of the specimen, might lead to inaccuracy (World Health Organization, 1999). Data from Tomlinson and colleagues indicate that two proprietary disposable slides (Microcell, Conception Technologies, San Diego, Calif; Leja, Leja Products, BV Nieuw- Vennep, The Netherlands) can result in a lower concentrations of sperm compared to the hemocytometer method (Tomlinson et al., 2001) . In contrast, plenty of reports document unacceptable differences between different laboratories and even between different individuals, although fewer studies attempt to address these issues. So, what is wrong? Improvement of semen quality testing has been emphasizing in some reports (Jorgensen et al., 1997; Keel et al., 2000). But due to low number of sperm evaluation by the conventional method results in poor reproducibility. These problems might be overcome when using flow cytometry. The validation of method is a challenge due to its essentiality of having specific, precise, objective, and accurate evaluation to establish a correlation of fertility data or to predict potential of a semen sample accurately (Amann, 1989). In a fertility clinic, precision of data in important as the result of semen analysis is frequently used to manage fertility of a patient and treatment of the unfertile couples. Thus, it is important to take into consideration within and between laboratory variations for successful infertility treatments. Sometimes its a matter of argument that compared to flow cytometry, fluorescent microscopy evaluate â€Å"patterns of fluorescence rather than the fluorescence intensity. Flow cytometer has the lack of ability to discriminate sperm containing a fluorescent marker bound to the equatorial segment or over one of the acrosomal membranes (Parinaud et al., 1993; Mortimer and Camenzind, 1989; Mortimer et al., 1987). Tao et al. (1993) compared flow cytometry and epifluorescent microscopy with various lectins and indicated that there is almost no difference between methodologies for detection of the acrosome reaction. However, it has been argued that lectins do not bind specifically to the acrosomal region of the sperm (Purvis et al., 1990; Holden and Trounson, 1991) and that other binding sites can be easily distinguished by epifluorescence microscopy, whereas flow cytometry identifies the signal from the entire sperm. Additionally, conventional light microscopic semen assessment is increasingly being replaced by fluorescent staining techniques, computer-assisted sperm analysis (CASA) systems, and flow cytometry (PenËÅ"a et al., 2001; Verstegen et al., 2002). Additional advantages over existing techniques are that this approach is faster than the hemacytometer and that cellular debris, fat droplets, and other particulate material in extended semen are not erroneously counted as sperm, as often occurs with electronic cell counters. This method can also be used to determine the number of somatic cells in a semen sample. Application of flow cytometry for sperm count Sperm count is an important predominant factor for the evaluation of sperm fertility potential. Accurate determination of sperm cell concentration is critical especially in AI industry because it provides assurance to customers that straws of extended semen contain the sperm numbers indicated which will help to decide appropriate doze especially for pig. Accuracy of sperm count is a common problem in the andrological laboratories and accurate measure of sperm concentration is particularly important for export in which verification of numbers may be required. Routine sperm counts can help to identify possible processing errors within a specific batch of semen or on a particular day, should those errors occur. As sperm counting procedures become more refined, routine counting can be used to monitor subtle changes in daily semen processing that might affect the number of sperm packaged in a straw. Every time new and more accurate methods for the sperm count determinations are coming and being replaced by the older ones. Some laboratories are trying the Maklerm counting chamber (Se if- Medical, Haifa, Israel) and other improved hemacytometers, such as the MicroCellTM (Fertility Technologies, Inc., Natick, MA); however, these techniques will likely have standard lems similar to those associated with the standard hemacytometers. Although hemacytometers are routinely used for sperm counts, due to the slow process and need for multiple measurements of each sample, the chance of error increase. Freund and Carol (ref) stated that a difference of 20% were not unusual between the determinations by the same technician. Electronic counters provide much more rapid counting, are easier to use, and give more repeatable results among technicians. However, those instruments tend to include in the sperm count any somatic cells present, immature sperm forms, cytoplasmic droplets, debris, and bacteria, thereby inflating the concentration value (Ref). Spectrophotometer is recently being used in the AI industries to assess sperm concentration by determining turbidity of a semen s ample using an instrument previously calibrated for sperm concentration with a hemacytometer or Coulter counter (Ref). The accuracy of this method depends on the methods used for spectrophotometer calibration. Although, sperm concentration can also be determined by spectrophotometrically, the debris present in the raw semen crease problem with misestimation. Sperm number in the frozen thawed semen is difficult to ascertain as most of the extender contain egg yolk particles, fats and other particles which affect measurement of sperm with electric cell counter or spectrophotometers (Evenson et al., 1993). On the other hand flow cytometry created possibilities of a rapid determination of sperm number in a precise form. It is the flow cytometry which can reduce intra-laboratory and inter-laboratory variation and conflict regarding sperm concentration assessment. Computer assisted semen analyzer is robust technique for analyzing sperm movement which can count sperm as well; but such an a nalyzer most of the cases use some counting chamber or hemacytometer which itself can generate error. Although, hemacytometer was originally developed for blood cell counting, its use is now diverse including andrological laboratories for sperm counting. Around two-decade ago flow cytometry was suggested for sperm numbers in straws of cryopreserved bull semen. Christensen et al. (-) observed similar results for sperm count with flow cytometry and hemocytometer for a number of species. Now a day a simultaneous determination of sperm viability and sperm concentration is possible which can avoid the chance of occurring differences between ejaculates leading lack of coordination with field fertility and laboratory analyses. Thus the present technology is more precise which can get rid of variation from handling the sperm sample and variation from pipetting and the analysis itself. Further, Prathalingam et al. (2006) concluded that there is similarities for sperm count result between flow cytometry and two newly approached method (image analysis and fluorescent plate reader) for sperm counting. Though, use of fluorescent plate was emphasized due to low cost and allowing large number of cells counting from a large number of ejaculates. Although flow cytometry has become a valuable instrument for andrological determinations, it is also blamed that sperm concentration by flow cytometry signify a higher value than the real one. The possibility arise might be due to that semen samples often contain some alien materials such as immature germ cells, epithelial cells, blood cells, cytoplasmic droplet, cellular debris etc. In the same way, frozen semen has higher chance to introduce such material as they contain diluents components especially egg yolk particles. These particles and cell debris might have frontal and side light scatter parameters those are similar to spermatozoa. Such sperm-count-overestimation problem arisen in our cases also, especially when we deal with frozen semen. Further it is also claimed that flow cytometry has a tendency to overestimate viable spermatozoa. We are also experienced with such trouble which we guess might be due to that egg particles of extender are considered as viable cell as for it s staining pattern. Our yet to publish data indicate that this problem can be mimic by a centrifugation process and by using low concentration sample for evaluation with flow cytometry. Very recently Petrunkina and Harrison (2009) proposed a mathematical equation for fixing this flow cytometric sperm counting. Thus much research is going on and we hope such discrepancy will completely be resolved near future to get advantage from this robust technology for sperm counting. Flow cytometry for detecting sperm intactness -Viability of spermatozoa The viability of spermatozoa is a key determinant of sperm quality and prerequisite for successful fertilization. Viability of spermatozoa can be assessed by numerous methods, but many are slow and poorly repeatable and subjectively assess only 100 to 200 spermatozoa per ejaculate. Merkies et al. (2000) compared different methods of viability evaluation. They concluded that Eosin-nigrosin overestimate viability while fluorescent microscope and flow cytometry estimate similar trend of viability. Current flow cytometric procedures are able to simultaneously evaluate sperm cell viability together with some other attributes. This method has been successfully used for assessing spermatozoa viability in men (Garner and Johnson, 1995), bulls (Garner et al., 1994; Thomas et al., 1998), boars (Rodrà ­guez-Martà ­nez, 2007; Garner and Johnson, 1995; Garner et al., 1996), rams (Garner and Johnson, 1995), rabbits (Garner and Johnson, 1995), mice (Garner and Johnson, 1995; Songsasen et al., 199 7), poultry and wildfowl (Donoghue et al., 1995; Blanco et al., 2000) and honey bees (Collins and Donoghue, 1999; Collins, 2000) and in fish (Martin Flajshans et al., 2004). Considerable information has accumulated on the use of fluorescent staining protocols for assessing sperm viability (Evenson et al., 1982). The SYBR 14 staining of nucleic acids, especially in the sperm head, was very bright in living sperm. Good agreement was observed between the fluorescent staining method and the standard eosin-nigrosine viability test; the flow cytometric method showed a precision level higher than that of the manual method. One of the first attempts to assess sperm viability utilized rhodamine 123 for determining potentiality of mitochondrial membrane while ethidium bromide for membrane integrity through flow cytometry (Garner et al., 1986). Other combinations that have been used to examine the functional capacity of sperm are carboxyfluorescein diacetate (CFDA) and propidium iodide (PI) (Garner et al., 1988; Watson et al., 1992); carboxydimethylfluorescein diacetate (CMFDA), R123, and PI (Ericsson et al., 1993; Thomas and Garner, 1994); and PI, pisum sativum agglutinin (PSA), and R123 (Graham et al., 1990). The most generally used sperm viability stain combinations is SYBR-14 and PI at present. This stains are now sold commercially as live/dead viability kit. When these two stains are combinely used, the nuclei of viable sperm take fluoresce green and membrane integrity lost cells take red stain. This staining technique has been used in a number of species, including the boar (Garner and Johnson, 1995; Saravia et al.,2005, 2007,2009). Although species differences do exist in the function of spermatozoa, the Live/Dead stain may similarly have no adverse affect on fertilization in the equine, although it remains to be tested in this species. Recently a new instrument (Nucelocounter-SP100) has been introduced to evaluate sperm concentration [11] and viability. Due to the small size and low cost, this instrument has been attracted for field measurements of both concentration and viability. In our hand this instrument was also became useful for the quick measurement of sperm concentration an d viability in stallion (Morrell et al., 2010). Fluorescent probes such as H33258, requiring flow cytometric analysis with a laser that operates in the ultraviolet light range, are less commonly used as this is not a standard feature on the smaller analytical machines. However, one alternative is to use a fluorometer. A fluorometer is a relatively low-cost piece of portable equipment that permits a rapid analysis to be carried out on a sample. Januskauskas et al. (2001) used H33258 to detect nonviable bull spermatozoa by fluorometry and obtained an inverse correlation between the damaged cells per cent and the field fertility. Another option is fluorescent attachments for computer-assisted semen analysis devices. For example, the IDENT fluorescence feature of the Hamilton-Thorne IVOS permits staining with H33258 allowing an assessment of sperm viability to be made along with motility. Fluorochromes used to assess sperm viability by both approach could be utilized in combination with each other. In that case, when CFDA is used combined with PI, three populations of cells as live, which are green; dead, which are red; and a third population which is stained with both and represents dying spermatozoa can be identified. This combination was found useful by Almlid and Johnson (1988) for frozen-thawed boar spermatozoa for monitoring membrane damage at the time of evaluation of various freezing protocols. Further, Harrison and Vickers (1990) also noticed that this combination with a fluorescent microscope is effective indicator of viability of fresh, incubated or cold-shocked spermatozoa in boar and ram. Contrasting to these, Garner et al. (1986) was failed to find a relationship between bull sperm viability and fertility when using combination of CFDA/PI . Flow cytometry for evaluating sperm viability appears to be a precious tool in the AI industry. When a high number of sperm is packed in each insemination dose, the effect of selecting the best ejaculates according to sperm viability has a relatively limited effect. However, sperm viability might be more important when combined with low-dose inseminations. The FACSCount AF flow cytometer also determines sperm concentration accurately and precisely during the same analysis (Christensen et al., 2004a). The combined assessment of sperm viability and concentration appears to be useful in the wake of improving quality control at AI stations. Because of the results of this trial, this method has been implemented by Danish AI stations (Christensen et al., 2005). Relatively bright fluorescence was found also in the mitochondrial sheath of living sperm. But the mechanism and mode of action by which SYBR-14 binds to the DNA of sperm is not known. It is know that PI stains nucleic acids by inte rcalating between the base pairs (Krishan, 1975). Viability stains can also be used in conjugation with fluorescently labeled plant lectins for simultaneous assessment of the plasma membrane integrity and the acrosome integrity (Nagy et al., 2003). It is conceivable that assessment of viability using SYBR-14 dye does not damage spermatozoa, since Garner et al. (5) found that insemination of boar sperm stained with SYBR-14 did not compromise fertilization or even the development of flushed porcine embryos in vitro. Non-viable sperms can be detected using the membrane-impermeable nucleic acid stains which positively identify dead spermatozoa by penetrating cells with damaged membranes. Plasma membrane which is intact will not permit these stains entering into the spermatozoa and staining the nucleus. Most frequently used stains include phenanthridines, for example propidium iodide (PI; (Matyus, 1984) ethidium homodimer-1 (EthD-1; (Althouse et al., 1995), the cyanine Yo-Pro (Kavak, 2003) and the bizbenzimidazole Hoechst 33258 (Gundersen and Shapiro, 1984). After a series of comparison between fertility of cryopreserved stallion spermatozoa with a number of laboratory assessments of semen quality as assessed by flow cytometry using PI, Wilhelm et al. (1996) concluded that viability is the single laboratory assay that correlated with fertility. -Sperm plasma membrane integrity Although the sperm plasma membrane covers the entire cell, it consists of three distinct membrane compartments, one which covers the outer acrosomal membrane, one which covers the post acrosomal portion of the sperm head, and one which covers the middle and principal pieces. Sperm membrane is directly or ind