[Cytometry] Teaching Flow Cytometry

Mario Roederer roederer at drmr.com
Sun May 8 14:33:54 EDT 2011


When I give instructional lectures on Flow Cytometry, I usually tell people: "Unfortunately, there is no RIGHT way to do a FACS experiment... but there are a whole bunch of wrong ways."  A corollary of this is that, no matter what, the experiment you just performed was done incorrectly.

The point is that you need to do each experiment multiple times, using variations, and to not treat the flow cytometer as a simple "input/output" device.  Experimental and instrumentation variables WILL affect your results, and you need to be sure that your conclusion is not derived from those variables as opposed to true biological processes. 

The complexity of flow cytometry engenders a desire to skip over all the details and just believe the output number.  Rather than repeating experiments after perturbing conditions to test effects, people all too often will simply assume the value is final and correct.  While borne of the inadequate time we have to learn new (complex) technologies, it manifests as an experimental laziness that will lead to the path of bad science.  I've witnessed the entire gamut of outcomes from this process: from publications that were not reproduible, to those that were retracted, and finally, to seeing careers ended following research misconduct inquiries.

Ultimately, flow cytometry is a very complex technology.  No, it's not brain surgery (<http://www.youtube.com/watch?v=THNPmhBl-8I>) ... but the shear number of variables that can directly impact the output measurement, sometimes in extremely subtle ways -- makes it daunting.  There is no substitute for experience -- and that's the other thing I tell people:  Don't be afraid to get help!  Even when you "know" the answer!

And that's perhaps the most important lesson of all:  nobody, even those of you who teaching cytometry in the 1960's, is done learning about flow cytometry and how to do it better.

mr


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