RE: tracking cell division, not CFSE

From: William King (wking@cnmc.org)
Date: Wed Jul 10 2002 - 15:10:16 EST


I've had luck with Sigma's PKH67-GL though it took some amount of time for me to
determine that the package insert's range is toxic. I seem to recall that a 40-fold
dilution did the trick without cellular toxicity and with sufficient labeling to
be bright.

William


C. William King, Jr.
Technical Coordinator Flow Cytometry
FACS core research facility
Children's Research Institute, Room 5220
Children's National Medical Center
111 Michigan Ave., NW
Washington, District of Columbia, 20010
202-884-2308
202-884-5808 fax
wking@cnmc.org


<<< "Reed, Doug S Dr USAMRIID" <Doug.Reed@DET.AMEDD.ARMY.MIL>  7/10  4:17p >>>
PKH-26 is also fairly toxic to cells, at least in our hands. That's why we use CFSE
instead.

-Doug

Douglas S. Reed, Ph.D.
Microbiologist
Respiratory & Mucosal Immunity
Department of Aerobiology & Product Evaluation
Division of Toxinology & Aerobiology
U.S. Army Medical Research Institute of Infectious Diseases
1425 Porter Street, Fort Detrick
Frederick, MD 21702-5011
301-619-6728
301-619-6911 fax
doug.reed@det.amedd.army.mil


-----Original Message-----
From: Joanne Lannigan [mailto:joannelannigan@virginia.edu]
Sent: Monday, July 08, 2002 10:15 AM
To: cyto-inbox
Subject: RE: tracking cell division, not CFSE



Hi Simon:
You could try PKH-26 a lipophilic dye which excites at 488 and emits in
the red/orange range. It is often used for cell tracking /proliferation
assays. I believe it is sold by Sigma.
Regards,
Joanne

Joanne Lannigan, MS
Director, Flow Cytometry Core Facility
Jordan Hall, Room 7067
P.O. Box 800734
Charlottesville, VA 22908-0734
Office: 434-924-0274
Lab: 434-263-2695
Fax: 434-982-1071
email: joannelannigan@virginia.edu

> -----Original Message-----
> From: Simon Monard [mailto:smonard@trudeauinstitute.org]
> Sent: Tuesday, July 02, 2002 2:31 PM
> To: Cytometry Mailing List
> Subject: tracking cell division, not CFSE
>
>
> Hi folks
>
> Has anyone had any success using both CFSE and another dye to track
cell
> division of
> two ex-vivo labelled cell populations.We use CFSE all the time and it
> works great,
> what we need is another orange or red fluorescing dye that like CFSE
will
> be retained
> by cells in vivo and will allow  us to follow the number of cell
> divisions. We have
> tried Snarf but it seems to be toxic to the cells we are using.  The
other
> dye must
> be excitable with either 488nm or 633nm. Any suggestions?
>
> best wishes
>
> Simon
>
> Simon Monard
> FACS Lab Manager
> Trudeau Institute
> Saranac Lake
> NY12983
>
> Ph 518 891 3080 X352



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