[Cytometry] Teaching Flow Cytometry

Mark Simmerson Mark.Simmerson at sch.nhs.uk
Mon May 9 07:34:34 EDT 2011

Hi All,
This has promoted some very interesting points.
I have the task of introducing flow to new people at a small clinical lab dealing with paediatric cases.
I use our competency assessment as a basis for teaching.
I start with a potted history of flow, what a flow cytometry is and what flow cytometry means. Although my users are not expected to set up the instrument from scratch, demonstrate compensation, etc. to show 'crap in, crap out'.
Things I have learnt.
1 - everyone has different learning styles.
2 - some show more aptitude for flow than others.
3 - some will not read the methods, some will read some of the methods and some will read the methods but not follow them. 
4 - it pays to be very, very patient.
5 - don't introduce material that users don't need - for our lab this means showing staff how to do single platform pipetting for CD34s, etc, perform and interpret acute leuk data, but not how to do compensation. Show staff what the data should look like and what to do if it doesn't.
I always try to remember back to my introduction to flow at the old lab at the Northern General Hospital in Sheffield, where Viv Grainger and particularly Ian Storie showed the patience of Job with me. I simply couldn't 'get' flow, no matter how they explained it.
Then one morning, I had a 'road to Damascus moment', a light went on in my head and verily I understood. But I don't think Viv ever quite forgave me for throwing away about 50 tubes he'd set up for a experiment he was doing...
We flow geeks can bang on and on about our little pet subjects, so for teaching -  keep it simple, keep it short and keep it relevant.
Mark Simmerson,
Senior BMS
Dept Paediatric Haematology,
Sheffield Children's Hospital.

>>> Mario Roederer <roederer at drmr.com> 5/8/2011 7:33 pm >>>
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.

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