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Below is a summary of responses for my previous question about dyes for yeast.  Thanks to all who responded.
 
I have a user who would like to stain / quantitate marine yeast in seawater samples.   Seawater is typically loaded with lots of bacteria so I am looking for an antibody / dye that will not cross react with bacteria.  I may be able to gate a lot of bacteria out by Forward / Side Scatter but there will probably still be an overlapping population with the marine yeast.
 
Thanks for your help.
 
Jayme U. Newell
SOEST Flow Cytometry Facility
University of Hawaii at Manoa
 
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-----Original Message-----
From: Jayme Newell [mailto:jnewell@pbrc.hawaii.edu]
Sent: Thursday, October 06, 2005 6:59 PM
To: Cytometry Mailing List
Subject: dyes for yeast flow cytometry.
 

Is there a standard dye used for staining of yeast cells for flow?
I've tried the FUN1 stain (a dye which fluoresces red for live cells and
green for dead cells) without much success.  I was not able to
compensate red / green.
 
I will post a summary of responses.
 
Thank you.
 
Jayme U. Newell
SOEST Flow Cytometry Facility
University of Hawaii at Manoa
 
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Jayme,
 
You don't specify what you'd like to stain for -- if you're looking for
yeast cell cycle/DNA content, I routinely use Sytox Green
(Invitrogen/Molecular Probes) with great success. 
 
Feel free to contact me for more information/staining protocol if this
is what you are interested in -
 
Regards,
Christina DeCoste
===========================================
Christina J. DeCoste
  Manager, Flow Cytometry Core Facility
Princeton University
Department of Molecular Biology
Lewis Thomas Laboratory
Princeton, NJ  08544
Phone: (609)258-1695
Fax:   (609)258-8468
cdecoste@molbio.princeton.edu
============================================
 

-----------------------------------------------
 
 
Dear Jayme,
 
 
 
The company Anomeric has a stain called Fungalase. It’s not suitable to differentiate between live and dead cells, and I’m not sure if it’s still available, though. The link is http://home.earthlink.net/~anomeric/fungalase.html
 
 
 
Hope this helps.
 
 
 
Arne
 
 
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Dear Jayme,
 
As you may notice, the staining of yeast cells is difficult doe to
the cell wall.
You cannot use current dyes easily because it very much depend of the
stainability.
 
For exemple, mitochondrial mmebrane potential dyes works in general
but needed long incubation time and a strict whasing procedure (this
is true for DiOC6(3) and other dyes).
 
The best but time consuming solution is to obtain sphéroplastes of
quality, because they have the same behaviour than mammalian cells.
 
Be also carefull because many publications are wrong and describe
very often artefacts...
Especially, when describing cell death processes... this being
especially the case of some of the F. Madeo and KU Frohlich
publications.
 
I am ready to help you if needed with more precises questions.
 
Patrice
 
Dr. Petit Patrice X.
INSTITUT COCHIN
INSERM U.567, CNRS UMR 8104, IFR 116
Department of Genetic, Development and Molecular Pathology
Team 5 "Cancer, Apoptosis and Mitochondria"
CHU Cochin Port-Royal
24, rue du Faubourg Saint-Jacques
F-75014 Paris, France.
 
Tel: 33 01 44 41 24 11
Fax: 33 01 44 41 24 21
E-mail: pxpetit@cochin.inserm.fr
http://www.cochin.inserm.fr
 
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Dear Jayme,
 
When I was sorting brewer's yeast we just used Propidium Iodide (PI), for live/dead.  Or H33342 for nuclear size.
 
Regards
 
Rob W.
 
 
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Dear Dr. Newell,
 
I have attached a poster that give you some other options for yeast
vitality by flow cytometry. 
 
FUN1 has localized red staining in live yeast, so is difficult to use on
a flow cytometer. However, you might try a red/green ratiometric
parameter with the FUN1 data to see if you can  tease out populations.
 
Best regards,
Bill
 
William Godfrey, Ph.D.
Flow Cytometry Research Area Manager
Invitrogen Corporation
29851 Willow Creek Road
Eugene, OR 97405
tel: 541-335-0141
toll free: 800-438-8300, x50141
mobile: 541-729-5316