Scatter change on fixed cells

From: Axl.Neurauter@dynal.net
Date: Mon Jul 10 2000 - 06:28:36 EST


We fix cells in PBS with 1 % paraformaldehyd and analyze these cells on a
four color Coulter XL.
The cells change forward and side scatter after about 24 hours (both
scatter decrease), and after that the scatter is stable for several days.
This change in scatter on fixed cells is not observed when the same cells
are analyzed in a FACS Calibur.
The volume-profile of the fixed cells is not changed as observed in a
Coulter Counter.

Has any one seen the same phenomena and could we get an explanation? Why do
we see a change in refractive index in the Coulter but not the FACS
machine?

Best regards
Axl Neurauter
Dynal Biotech, Immunosystems R&D



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