[Cytometry] Teaching Flow Cytometry

Christopher A. Worth caw at bcc.louisville.edu
Mon May 9 14:39:34 EDT 2011


Actually it isn't just the so-called "new" people.  The manufacturers are dragging us down the magic juice/magic box path.

to sum up

1) power on

2) put sample into preprepared juice

3) run auto calibration sensing setup alignment optimization bead cocktail.

4) put on tube press go

5) get automated printout  give to clinician that requested the test

6) repeat.


Now get a high school student to work part time running your samples. :)



I get people that run a calibur.  They come to me when they're ready to sort.  They don't have any idea what is going on inside the box aside from "we collect FL1 and 
FL2 and FL3"


and to address Till's question

I start with the basics.  Just what is going on from the time you put the tube on until you get data or sorted samples out.  Most of my customers understand the staining 
concepts already, from having done analysis to get where they want to sort.







On Fri, 6 May 2011 21:25:06 -0400, thomas delohery wrote:

>I disagree.  Howard's comments fit perfectly with the vast majority of "new people" I have encountered in both industry and academia over the past 10 years.  Howard's 
comment was not directed at Till but to his potential students; as such, his comments are perfectly appropriate.

>TomD

>On May 6, 2011, at 11:46 AM, Facs wrote:

>> Howard wrote:
>> 
>> "Problem number one these days seems to be motivating the new people to think; too many of them want to mix samples with magic juice, put them in the magic box, 
press the magic buttons on the box and do the necessary magic mouse clicks, keystrokes, etc. on the computer, and save the PowerPoint presentation and the 
manuscript that will be submitted online to one prestigious journal or another. Then it's back to social media."
>> 
>> I think you should rethink this one as the response is not relevant to the new "motivated" individual who posted the question. 
>> 
>> Ann Atzberger
>> Flow Cytometry Facility
>> Institute of Molecular Medicine
>> Trinity Health Sciences: St James Hospital
>> Trinity College Dublin
>> Dublin 8 Ph. 0035318963055
>> ________________________________________
>> From: cytometry-bounces at lists.purdue.edu [cytometry-bounces at lists.purdue.edu] On Behalf Of Howard Shapiro [hms at shapirolab.com]
>> Sent: 06 May 2011 14:52
>> To: Dettmering, Till
>> Cc: hsfc at elist.tufts.edu; flow list Purdue flow list Purdue  (cytometry at lists.purdue.edu)
>> Subject: Re: [Cytometry] Teaching Flow Cytometry
>> 
>> Till Dettmering wrote:
>> 
>>> I'm a PhD student for a year now and I'm working with flow cytometry for 2.5 years. Since there is no real dedicated FACS-'guru' in our group who I could have 
learned from, I had to teach practically everything about flow cytometry to myself, including maintenance of the device (a Partec PAS III). I find myself more often in the 
situation in which I have to teach new students who have never seen a flow cytometer how to work with it and how to interpret the data. Although it works out quite well, I'm 
very interested on your experiences on how to best teach new people flow cytometry. What do you start with? Do you have something like curriculum you follow when 
teaching it? What did you start with when you learned it? What should one pay special attention to when teaching?
>>> 
>>> I'm very interested in hearing your experiences!
>>> 
>> 
>> Problem number one these days seems to be motivating the new people to think; too many of them want to mix samples with magic juice, put them in the magic box, 
press the magic buttons on the box and do the necessary magic mouse clicks, keystrokes, etc. on the computer, and save the PowerPoint presentation and the 
manuscript that will be submitted online to one prestigious journal or another. Then it's back to social media.
>> 
>> Those of us who write flow books and vent on this Mailing List have tried to spell out the basics. Cytometry, as I have often said, begins with the cell, even at the level 
of the word itself. You have to know what information you want from the cells in your samples, and what reagents you need to use and parameters you need to measure 
to get that information, and it helps to consider alternatives when they are there.
>> 
>> The cell identification game is like "Where's Waldo (his name is Wally in some places)?" - The game wouldn't be any fun if Waldo were the only person in the crowd 
wearing magenta and chartreuse; the intellectual challenge comes in picking out physical features and the patterns of the colors on his clothing and distinguishing them 
from those on everybody else in the crowd among which he is hidden. Generally speaking, one can always identify a cell from a given species by detecting a specific 
nucleic acid sequence, or a combination of antigens. In unlysed blood, CD45 is good for discriminating white blood cells from red cells but, in cases where you know you 
won't encounter nucleated red cells, it's much simpler to use a permeant DNA dye. Simpler is always better.
>> 
>> One can learn these kinds of tricks from books and material available online. It's harder to learn how to set up an instrument, verify that it is working properly, and deal 
with it if it isn't. The theoretical part of that is accessible in books, but which knobs you turn and/or how you set up the software depend entirely on the specific cytometer 
you're using. Manufacturers offer training courses, and provide manuals, but, as is the case with computers, you can sometimes RTFM and find that it doesn't really tell 
you all you'd need to set up a flow lab in a postapocalyptic venue. Oral tradition only goes so far; even prehistoric hunters seem to have needed seminars with 
presentations painted on cave walls.,
>> 
>> When TFM leaves you in the dark, you can get computer questions answered online, sometimes for nothing, or you can buy something like the books in David 
Pogue's "the missing manual(R)" series, published by O'Reilly.
>> 
>> What we need in cytometry are equivalents. Practical Flow Cytometry had its genesis in my "Building and Using Flow Cytometers: The Cytomutt Breeder's and 
Trainer's Manual," and the material from that on building and using instruments, which appeared in the first couple of editions of PFC, has apparently been sufficient to 
enable dozens of people to put together instruments and keep them running for decades without access to service from outside.
>> 
>> It does not strike me as unreasonable that manufacturers should support their user groups in generating similar documentation, but the users could get organized to 
do this without the manufacturers' help, which would minimize the likelihood of problems being swept under the rug. The material, which should cover software from both 
manufacturers and third parties as well as apparatus, should be kept online in a presumably neutral site such as this one.
>> 
>> I've seen people with little or no lab experience get up to speed, at least with relatively simple flow cytometers, in weeks, and people with decades of experience fail 
to do so in months to years, and I can't always explain either.  There hasn't been a Mozart in cytometry yet, and even he started small. If you can't hear a sonata playing in 
your head, don't try for a symphony, and, if you can't do two-color fluorescence and two-angle scatter, don't try running a 4-way sorter with 7 beams and 24 colors. I spent 
several years training in surgery, and would have had to spend several more before anybody let me do any significant part of a heart operation; it is, admittedly, harder to 
kill people by running a flow cytometer incompetently than it is to do so when you are waving a scalpel around vital organs, but the malpractice of adding junk to the 
medical and scientific literature has its own adverse consequences.
>> 
>> So let's get organized.
>> 
>> I'll be at my 50th College Reunion during ISAC, which declined my offer to give a plenary talk by remote video, but catch up to me online if you're willing to help.
>> 
>> -Howard
>> 
>> 
>> 
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