RE: Mast cells and basic orange 21

From: Weaver, James L (WEAVER@CDER.FDA.GOV)
Date: Mon Oct 28 2002 - 13:40:07 EST


You might take a look at: Perretti-M et al (1990) Investigation of rat mast
cell degranulation using flow cytometry. J. Pharmacol. Meth. 23 187-194.
They found that using light scatter alone tracked histamine release
reasonably well. The plots in the paper show that the highly granular rat
peritoneal mast cells have a very high SSC signal. After degranulation, the
cells drop out of this gate. You might try this with your cell line.

-Jim Weaver


*************************************************

 James L. Weaver Ph.D.
 Division of Applied Pharmacology Research
 Office of Testing & Research
 CDER MOD-1, FDA
 8301 Muirkirk Rd, Laurel MD 20708

 Phone: 301-827-8237
 Fax: 301-594-3037
 Email:WEAVER@CDER.FDA.GOV

 This email is my personal communication and
 is not in any way official U.S. FDA Policy
*************************************************



*  -----Original Message-----
*  From: Amy Raber [mailto:araber@athersys.com]
*  Sent: Friday, October 25, 2002 2:58 PM
*  To: Cytometry Mailing List
*  Subject: Mast cells and basic orange 21
*
*
*
*  Dear Flow users,
*
*  I am interested in stimulating a mast cell line and using
*  flow cytometry
*  to screen for degranulation.  I came across mention of a dye-basic
*  orange 21 that may be appropriate for this assay.  I was
*  hoping someone
*  would know where I can purchase this dye?  Is there a
*  protocal somewhere
*  for staining?  Or is there a better method to measure degranulation?
*  (we are looking into annexin staining for degranulation too)  We only
*  have access to 488nm laser.
*
*
*  Thanks so much for any help you can send my way!!
*
*  Amy
*
*  Amy Raber
*  Athersys Inc.
*  Cleveland, OH
*
*



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