Re: Simple flow cytometer construction

Gene Pizzo wrote:
> The second edition of Howard Shapiro's Practical Flow Cytometry has an entire section
on the construction and design of a flow cytometer and you may also gain by perusing some
of the historical data presented in the latest edition of Purdue's Cytometry DVD. If it
were me however I would think this is too extreme for high school students and I would
pursue funding to get a used Scan or Calibur with which I could demonstrate cytometry
techniques to these students. You'd then have a system up and running with which you
could reverse engineer some of the systems design involved in building an instrument as
well as demonstrating technique. People forget that in immunology there was a long period
of time
> in which cytometry wasn't being used even while the technique was available because it
was too expensive and cumbersome.
> ________________________________
>
> From: Patricia Lovelace [mailto:patricia.lovelace@sbcglobal.net]
> Sent: Wed 7/9/2008 18:40
> To: cyto-inbox
> Subject: Simple flow cytometer construction
>
>
>
> Hello flowers,
>
> A friend who left the world of cytometry to become a
> high school teacher is asking if there is a resource
> he could look into to give ideas about constructing a
> simple working flow cytometer from readily available
> items. He is looking into incorporating such a project
> into his science lesson plans. Does anyone out there
> have any ideas of where he could look for information?
>
>
> Can it be done?
>   
Yes, it can be done. Wiley decided to drop the build-it-yourself section 
after the 2nd Edition of Practical Flow Cytometry, because the book sold 
about 6,000 copies and only about one per cent of the people who read it 
built flow cytometers. At least two of the Cytomutts ran for twelve 
years or more and were maintainable by the builders (with perhaps four 
phone calls to me in the interval). There hasn't been a commercial flow 
cytometer with that good a service record yet!

Wiley gave me permission to copy and circulate the relevant chapter 
(Building Flow Cytometers) from the 2nd Edition, and anyone interested 
can get it from me (I added one page to suggest what changes might be 
made 20 years after the chapter was written) as a 4.4 MB .pdf file; just 
drop me an e-mail. I offer small amounts of free advice to people in 
nonprofits and small and larger amounts of paid advice to people trying 
to make money with flow cytometry, provided I find the people likeable. 
We may be devoting some time to bringing the Cytomutts up to date later 
this summer, and may put updates on the shapirolab.com web site, which 
could use some work.

Acquiring a used FACScan, FACSCalibur, etc. will cost you much more - 
probably at least three times as much - as building a multiparameter 
Cytomutt. PMTs cost money, but you can easily use cheap ones for scatter 
and for green and shorter fluorescence wavelengths. Electronics are 
cheap, and you don't need expensive lenses. Most of the optical filters 
you need can be bought from manufacturer's overstocks for tens of 
dollars. Red He-Ne and diode lasers are cheap and reliable; green YAGs 
tend to be less reliable but some suppliers provide better products. The 
bubble for 488 nm solid state lasers will burst later this year because 
Nichia is already producing a 488 nm 20 mW laser diode (the current 
solid state lasers are all doubled, and therefore much more expensive), 
so one would expect substantial price drops for this wavelength, which 
remains the bread-and-butter light source for flow. A red or green laser 
source system should be buildable for under, possibly well under, 
$10,000, including the computer. If you don't have LabView or MatLab, 
for both of which some flow packages have been written, software may be 
an issue.

Cytomutts have been made to sort, using relatively simple and cheap 
electronics, but I haven't documented these in a neat package because 
neither I nor most of the Cytomutt builders had a pressing need for sorting.

There are, of course, cytometric applications, ranging from microbiology 
to some immunophenotyping (e.g., CD4), for which a very simple, very 
cheap (buildable for well under $2,000) "cellular astronomy" 
fluorescence imaging cytometer is perfectly adequate. More details on 
that should appear in my next book, which I hope will appear in 2009.

-Howard