Re: refrigerated samples

Maggie Brown (mrbrown@nih.gov)
Mon, 29 Sep 1997 14:40:12 +0100

Keith Bahjat wrote the following:
>I only mention this as I have used BD's lysing solution for
>a very long time, and I have NEVER had it not adequately
>lyse 100 uL of peripheral anticoagulated (EDTA, Na Heparin,
>or ACD) whole blood when 2 mL of lysing solution is used
>UNLESS the sample was refrigerated.

We have had problems with BD FACS lyse when I know that the blood was never
refrigerated. There are just different instances when the red cells will
not lyse.

We have two lasers on our FACSort. Because we have the capacity of using a
fourth color in conjunction with our three color protocols, we have found
adding CD45 APC to the blood after staining and lysing to be a nice
solution to the problem of red cells resistant to lysing. By using a
combination of a light scatter gate and CD45 bright histogram gate we get
excellent results. (This will work until we move from routinely using
three colors to using four colors in our protocols).

Margaret R. Brown
NIH, Immunology, Clinical Pathology
Bldg10/2C410
Bethesda, MD 20892