You need a good cytometry book and attend one of the large cytometry
meetings. and examine the "guts" of the various cytometers. There are more
than Coulter and BD on the market. Don't forget about Partec and Apogee Flow
in the UK and Howard Shapiro custom builds his own version of cytometer..
The best intor book to cytometry is the latest edition of Flow Cytometry by
Howard Shapiro but I have a large Yellow book called Flow Cytometry and
Sorting ,2nd edition, Eds Myron R. Melamed, Tore Lindomo and Mortimer L.
Mendelsohn (Wiley-Liss/ISBN 0-471-56235-1). It has been an excellent
resource.
For light scatter, most cytometrist consider any light scattered greater
than 15 degree to be "side scatter" which is a general indication of
granularity of the cell and light scattered at less (usually much less than
15 degrees is an indication of size). The theory for this though is assuming
that your particle is completely spherical but with must eukaryotic cellls
this principle holds true but when you drastically deviate from this shape
like when analyzing bacteria or even reed blood cell which are really
biconcave disk, you will get several population depending on the shape of
the particle and the orientation that it was in when it intersected the
light source.
There is a really good chapter in the "Yellow Book" entitled "Light
Scattering and Cytometry" by Gary Salzman et al.
For the rest of your questions, the answers depend on numerous factors. You
have asked enough question for a book!!!
Good Luck
Tim Chance
CD
Atlanta, GA
-----Original Message-----
From: David Boyett [mailto:dboyett@axiombio.com]
Sent: Thursday, November 29, 2001 4:01 PM
To: cyto-inbox
Subject: RE: Flow Cytometer design
I am doing research on Flow Cytometers and wonder if anyone can offer
any insight. In particular, I wish to find out about compact flow
cytometers, and flow cytometers using micro fluidic chips. I want to find
out about flow paths investigated including free space, and channel confined
and how sheath flow is handled for each case. We are interested in cell
flow rates (minimum and maximum) and cell types and sizes under
consideration. I am further interested in system architectures,
particularly excitation wavelengths and excitation sources for those
wavelengths, but also geometries, tolerances, and signal to noise ratio
considerations. What fluorescent dyes are used, and what are their
absorption bands? How are micro spheres used in drug discovery with flow
cytometers? Especially what dyes, wavelengths, etc. are involved. I also
want to understand how many and which emission wavelengths are of interest
in flow cytometry for drug discovery. Finally, we are interested in
algorithms that use scattered light- forward and side- to determine cell
size and roughness. What specific angles must be used, how collimated and
coherent does the light need to be, and how are the data analyzed to
determine cell characteristics?
_________________________________________
David Boyett
Electro/Optic Engineer
Axiom Biotechnologies
3550 General Atomics Court
San Diego, CA 92121
858-455-4480
dboyett@axiombio.com <mailto:dboyett@axiombio.com>
www.axiombio.com <http://www.axiombio.com/>
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