Re: Help: FACScalibur problems

From: Daniel Vaulot (vaulot@sb-roscoff.fr)
Date: Fri Jan 31 1997 - 02:57:32 EST


Dear Gunnar

What stain are you using?  Stains like acridine orange used traditionally
for enumeration of marine bacteria by epifluorescence microscopy are
improper for flow cytometry.  The best to use is SYBR Green (see our recent
paper: 1997 Marie, D., Partensky, F, Jacquet S., Vaulot, D. Enumeration and
cell cycle analysis of natural populations of marine picoplankton by flow
cytometry using the nucleic acid stain SYBR Green I. Applied Environmental
Microbiology 63: 186-193.)

You should of course filter the sheath fluid through 0.2 umand use either
filtered sea water (best to match the refractive indexes of the sheath and
the sample) or Q water.
Pn the FACsort we typically use Green PMT settings of 500 and get very
little noise.

Good luck.


______________________________________
Daniel VAULOT
Station Biologique CNRS - INSU - UPMC
BP 74 
29682 Roscoff Cx FRANCE

Ph: 33 2 98 29 23 34 Fax: 33 2 98 29 23 24

e-mail: vaulot@sb-roscoff.fr
W3: http://www.sb-roscoff.fr (PhytoPlankton Group)



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