 |
 |
 |
 |
 |
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)
|
|
|
|
|
|
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