RE: fixing and storing cells
H.-G. Kreysch (kreysch@merck.de)
Thu, 13 Jun 1996 18:59:20 +0200
Although we don=B4t analyse cells under normal conditions after extended
storage, we have tested the stability of human PBMC which were stained with
FITC or PE conjugated antibodies and subsequently fixed and stored for up to
14 days in 1% paraformaldehyde/PBS (L.L. Lanier,N.L. Warner, J. Immunol.
Meth. 47:25-30 (1981)).
During this time period we could not find significant changes in the size of
lymphocyte subsets. Nevertheless, decreased FSC signals (which did not cause
problems for gating on lymphocytes) and variations of the absolute
fluorescence intensities of some antibodies tested in our lab were observed.
R.B. Lal et al. (Cytometry 9:213-219 (1988)) describe instabilities of
immunofluorescence stainings after prolonged storage of fixed cells which
may result from the antigen and/or the antibody or the fluorochrome.
Thus, I would strongly recommend to test each antibody / antibody
combination for stability under conditions used in your lab. =20
=20
=20
------------------------------------------------------------------------
H.G. Kreysch
Merck KGaA
Biomed Fo/IMO - A25/410 Tel.: (49)6151 - 72 7323=20
Frankfurter Str. 250 FAX: (49)6151 - 78 1332
D-64271 Darmstadt E-Mail: kreysch@merck.de
CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
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If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
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, EMAIL
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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web
http://www.cyto.purdue.edu
, EMAIL
cdrom3@flowcyt.cyto.purdue.edu
