hematopoietic cell sizing by flow

R.E.vanLeeuwen@tn.utwente.nl R.E.vanLeeuwen at tn.utwente.nl
Mon Oct 14 11:06:32 EST 1996

Dear Scott,

The forward scatter relation with size only holds for very small 
angles. This depends on the exact cytometer configutration (e.g. the 
kind of flowcell you use). The small angle forward scatter is related 
to diffraction, which is proportional to the cel/bead size.

The best thing to do if you are using a FACScan or likewise apparatus
 is to tape off the outer side of the forward scatter detector. Only scatter 
light around 2-5 dgrs. will be measured in that way. 
These angles are within the diffraction lobe of the forward scatter pattern.

You can do a bead experiment to see if the amount of taped off 
detector area is enough. As the highest intensity is in the small 
angles, no problems regarding signal intensity will occur.

Good luck.

Robbert van Leeuwen

Applied Optics / Flow cytometry Group
University of Twente
The Netherlands.

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