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Flow Cytometry: The Go-To Technology for Single Cell Analysis

“The global flow cytometry market is expected to reach $4.93 billion by 2021 from $3.14 billion in 2016, at a compound annual growth rate of 9.4 percent between 2016 and 2021. In the flow cytometry industry, market growth is majorly driven by the development of novel flow cytometers, increasing use of flow cytometry in clinical trials, launch of new reagents for specific applications like diagnostics and drug discovery, development of user-friendly and intuitive software, growing prevalence of cancer and HIV/AIDS, and growing adoption of flow cytometry techniques in research activities.” – Market Reports Hub, July 2016

J. Paul Robinson, Ph.D., professor of cytomics at Purdue University in West Lafayette, IN, and director of the Purdue University Cytometry Laboratories, also cites tremendous growth in the field and offers some reasons why. Robinson, along with John P. Nolan, Ph.D., principal investigator at the Scintillon Institute for Biomedical and Bioenergy Research in San Diego, CA, are instructors of two half-day SLAS2017 Short Courses – Introduction to Flow Cytometry and Advanced Flow Cytometry.

Why Flow?

“Flow cytometry is a technology that allows us to look at single cells, one cell at a time and look at many, many properties of those cells,” Robinson says. “Also, you can analyze the different properties of cells, accumulate them into populations and look at the differences between mixed populations in very effective ways.”

Flow cytometry research applications may include apoptosis, b-cell, bead-based immunoassays, cell and tissue microscopy, clinical, intracellular flow, multicolor flow, stem cell and t-cell immunology according to global medical technology company BD Biosciences. The SLAS2017 courses explore flow cytometry in high-throughput screening, advanced multiparameter analysis, spectral technologies, cell sorting and clinical and environmental (microbial) applications.

Robinson says one reason for the growth of flow cytometry is the vast number of reagents available now to phenotype the cells. This wasn’t the case in the early days of flow when the first fluorescence-based flow cytometry device (ICP 11) was developed in 1968 by Wolfgang Göhde from the University of Münster, Germany.

“In phenotyping, we ID cells with different properties,” Robinson continues. “Because we have these vast numbers of reagents now, and I mean in the thousands, we need to capitalize on them.” He adds that many of these reagents are monoclonal antibodies, which were developed specifically for flow cytometry use, another reason for industry growth. “An entire industry focused on development of monoclonal antibodies for every conceivable subset of cells that exist – not only for humans but in animals, plants, insects, everything. This is a big deal.”

More Companies Jump into the Game

In part because of the availability of more reagents, equipment manufacturers have retooled, adapted and acquired to meet the needs of basic researchers. John Joslin, research investigator at the Genomics Institute of the Novartis Research Foundation in San Diego, CA, discussed flow cytometry in high-throughput screening in a 2014 SLAS Electronic Laboratory Neighborhood e-zine article and SLAS Webinar.

"Flow cytometry is a very powerful tool that has been used for decades, and allows for multiparametric readouts at the single cell level within heterogeneous cell populations," Joslin states. "However, in the context of high-throughput screening, flow cytometry has traditionally been slow, low-throughput and not amenable to automation. The Novartis fully automated screening system solved that problem.”

Novartis responded to a need within its own industry. Robinson also points to those in life sciences research taking tools utilized in other fields and adapting them for use in basic life sciences research. Or even to big players in other fields seeing a hole they can fill effectively in the life sciences.

“For example, spectral analysis previously was not done in flow, but it was big in imaging companies,” Robinson says. “Now you can expect multiple companies to build these instruments. In fact, one of my patents is the foundation for spectral flow cytometry, and it was bought by a company that no one would have expected to enter the field of flow cytometry – Sony. When Sony licenses your patent, that has impact in the field. Now they’re a competitor to the traditional life sciences companies. An application that wasn’t in the field has opened up a competitive business. Sony is not going to be there alone. The company that drove spectral analysis in imaging in 2000 was Zeiss and they developed a new component – an add-on to their confocal microscope. Now, there’s no confocal microscope manufacturers that don’t have spectral capabilities – it became a standard offering in the field.”

Another area receiving a boost from the growth of flow cytometry is data processing, says Robinson.

“Whenever you have automation, you’re bringing large amounts of data together in a very fast, furious way and you have to be able to come out with intelligent ways of analyzing the data,” he says. “The field of flow is very quickly driving advanced data processing systems to handle the huge amounts of data with large numbers of parameters in very short periods of time produced by flow cytometry.”  

Cytometry for Life 

Robinson has experienced much in his career but 2006 was life-changing for this academic.

“I’ve been in this field my entire adult life and sometimes we become a little bit blasé,” he says. “We assume that everything that can be done is being done. You assume that the needs of a field are being met and I made that assumption. I assumed everybody else was solving the problems of low-cost devices in the field of HIV AIDS. I think that assumptions can lead to rude awakenings that sometimes aren’t pleasant.”

This happened for Robinson as president of the International Society for Analytical Cytology when they brought Stephen Lewis, Canadian advocate in the fight against HIV/AIDS in Africa, to speak at their conference.

“Lewis said quite emphatically to us that we were not solving the problem of HIV/AIDS in resource-limited countries,” Robinson notes. “He made me sit up. Here we were the experts in the field of CD4, and what were we doing for low-cost CD-4? Not much. Can we do it? Of course we can. I went home from that meeting really challenged by the fact that we were not doing what we should be doing. We were building big expensive instruments that were accommodating the needs of the western world and we were doing a great job of that. We could build the biggest, meanest, most expensive, most complex systems, but when you take those to resource-limited countries they were not working.”

Robinson’s response was to set up Cytometry for Life to bring together resources to build low-cost instruments that could be utilized in areas affected dramatically by HIV/AIDS but currently not being served.

“Boy, did I get a shock. We designed low-cost instruments and they worked. But do you think we could get any companies interested in building them? Of course not. I got the second rude shock of my life – if you build it, they will not come! I got so frustrated, I really couldn’t believe it. Generally speaking, in most things I’ve tried to do in life, I’ve been successful. I just couldn’t make this work. I had to back away – it’s a very sad story actually. I basically withdrew into my academic cocoon, my ivory tower, and said I can’t solve this problem.”

That is not where the story ends, however. “I sat down and told myself you’re not clever enough to solve the problem, you better go and think about how to reset your goal. I went off and did some crazy things – that I had no training in and no knowledge of – but I thought I really have to rethink the psychology of how I go about solving problems. That’s when I went off and ended up climbing Mt. Everest.”

Robinson said he took this drastic action to prove to himself that he could take up a challenge and figure out the tools and strategies needed to make it happen. The result? He summited Mt. Everest on May 23, 2009. With renewed hope, desire and faith in his abilities, he returned to Purdue determined to reset his goals for Cytometry for Life. 

“Now at Cytometry for Life, we are approaching a slightly different problem and teaming up with some experts in the area of malaria detection – a real problem in resource-limited countries. We are approaching the problem slightly differently than we did with HIV/AIDS by trying to build slightly bigger teams and involving more people who have expertise in the area. Among those people is one of the best known people in the field of flow cytometry – Howard Shapiro [Center for Microbial Cytometry in West Newton, MA Boston]. Howard wrote the textbook in the field and he’s now a key player in Cytometry for Life and integral to developing our new direction.”

SLAS2017 Short Courses

Introduction to Flow Cytometry is being held Sunday morning, Feb. 5. This course touches on the fundamentals and applications of flow cytometry across a range of life sciences applications. Participants learn about the historical development of the field – why it was invented, what problems it tries to solve, and more.

“We expect that participants of this course know nothing about the field of flow cytometry but have a great deal of knowledge in other fields like fluidics, automation or data processing,” Robinson says. “Perhaps their technology is now in competition with flow or they work in a company that now makes instruments or reagents for flow cytometry and wish to have a better understanding.”

Advanced Flow Cytometry takes place Sunday afternoon, Feb. 5. Here, instructors Robinson and Nolan dig deeper into leading-edge applications or expansions of current technology, including cell sorting; advanced data processing; spectral flow cytometry; analysis of extracellular vesicles and other nano-particles; high-throughput flow cytometry; hardware approaches, software and applications; and clinical, environmental and microbial cytometry. This course also provides participants with a strong background level of understanding of the technology, current limitations and the future opportunities.

Short Course Only Registration is available. Participants do not need to be registered for the full SLAS2017 Conference and Exhibition to attend a short course. The SLAS member registration fee for either of the flow cytometry courses is $325; non-members can register for $375; and students can register for $75.

December 19, 2016