StudiesFlow Cytometry What’s Your Motto? “Going with the Flow” or “Superflow?” Whether you think about flow cytometry as a “black box” or you can easily operate the most sophisticated of cytometers, nearly all scientists appreciate that flow cytometry provides a lot of information about one’s sample in a relatively short period of time. In a 27-question study, The Science Advisory Board asked over 750 researchers how they use flow cytometry to advance their research goals. Because flow cytometers come in many different configurations and at a wide range of price points, scientists must consider many factors when determining which brand of flow cytometer will best meet their needs. Complicating this decision is whether to perform these experiments in-house or outsource them to a flow core lab. This study addresses these issues from a scientist’s perspective by examining what critical factors affect both instrument and reagent use. Since the late 1970s, flow cytometry has enabled scientists to analyze a variety of cell types. Through optical means, flow cytometers distinguish cells on the basis of size and shape in addition to the presence of many different molecules inside and on the surface of the cells. Flow cytometry offers numerous advantages over other cell-based techniques: • Tremendous speed • Exquisite precision • Preservation of cell viability and cellular functions • Simultaneous measurements of multiple cellular parameters Applications The appeal of flow cytometry arises from the flexibility and sensitivity of fluorescence technology combined with the technique’s high speed and powerful data integration capabilities. Flexibility, sensitivity and speed enable this powerful technique to be used for multiple applications. In this study, cell sorting is the most common application of flow cytometry. Apoptosis, cell cycle studies and fluorescent protein detection rank closely together as the 2nd, 3rd, and 4th most popular applications of flow cytometry, respectively. Instrumentation Preference Respondents rely more heavily on cell analyzers as compared to cell sorters, (i.e., slightly more than twice as many tubes are analyzed versus sorted on a weekly basis). Given flow cytometery’s popularity, it was somewhat surprising that nearly 20% of respondents do not know what instrument brand they use. This lack of knowledge is likely due to the fact that 60% of study respondents utilize a flow core lab’s services. The degree of familiarity with a flow cytometer’s capabilities increases if respondents perform their flow cytometry experiments themselves—with those respondents working in a flow core lab being the most familiar of all respondents with their instruments. Of the scientists who are familiar with their instrument’s supplier, BD Biosciences is the most commonly used brand of flow cytometers. BD Biosciences possesses a large number of instrument features that respondents are relatively more satisfied with than features of other brands. Most respondents are the least satisfied with their flow cytometer’s ability to measure and/or monitor rare events—although this attribute varies slightly in its degree of importance by brand. Reagent Selection and Use Flow cytometer users from BD Biosciences, Beckman Coulter, and Dako have a strong affinity to use flow cytometry reagents from their primary instrument suppliers. Despite these preferences, there is little perceived differentiation among suppliers of flow cytometry reagents, however, Sigma-Aldrich is perceived to be slightly different. Reagents from BD Biosciences Immunocytomery Systems, BD Biosciences Pharmingen and Molecular Probes are the leading brands in the flow cytometry market. In aggregate, the majority of respondents select their primary brand of flow cytometry reagents based upon brand reputation (except for Santa Cruz Biotechnology). Performance is also an important factor in this decision (except for Dako). Additional factors also contribute to the choice of one brand over another and these considerations vary by supplier. These factors include favorable pricing, range of conjugated primary antibodies, and/or dye compatibility with other experiments. Future Directions Given that the majority of study respondents employ a core facility to conduct their flow cytometry experiments, it is not unexpected that a flow core lab’s recommendation is the most common factor that influences which flow cytometer brand a respondent uses. Nevertheless, brand reputation and performance are more significant reasons respondents use one instrument brand over another. The importance of both brand reputation and performance does vary by instrument brand. Given how versatile the technology is, most brands are well suited to carry out multiple applications. As experiments become increasing more complex in this post-genomic world, scientists are looking for instrumentation that can support their research and allow them to efficiently employ a variety of complimentary approaches and integrate several applications. Flow cytometry brands that address this need for managing the complexity of today’s experimental designs, will be better able to secure a place in the laboratory of the future. Conclusions In this study, there appeared to be a dichotomy between the types of researchers who preferred to use the more sophisticated flow cytometers and those that are happy with standard instrumentation. This first group of scientists professed a keen interest in a hands-on approach to data generation and analysis and did not seem to mind the amount of time it might take to master such a complex instrument: “The instruments should have highest possible capabilities such that I can decide which questions I want answered using this instrument. It's unacceptable to withhold my questions because of an instrument's limitations.” -Research Assistant, Europe “While complexity is not always the highest mark for usefulness, the potential to derive the most information from a given set of samples is more appealing than obtaining a set of sure results from a machine with only limited capacities.” -Postdoctoral Fellow, North America While technology drives innovation, not all scientists are interested in the most advanced flow cytometry models. Time concerns, limited resources and a lack of confidence in one’s technical abilities motivates many of these researchers. “I'm sure I'm not the only scientist around with a top of the line piece of equipment on the floor that's only being used for the most basic of purposes. Most of the time it's because we have only the vaguest idea of how to use it to the best of its capability. It's amazing what sort of experiments we can dream up and do with the most limited and archaic of tools sometimes!” -Research Assistant, North America “My use of a flow cytometer does not require a top-of-the-line machine and I prefer to take advantage of an experienced technician and have less headaches.” -Professor, North America To read more opinions from your fellow researchers about the advantages and disadvantages of different types of flow cytometers, please access this page. [ View Current & Future Studies ] [ View Past Studies ] |
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