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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