Tracy, If you could not find an effective way to quench the autofluorescence, you may consider the method of autofluorescence compensation. You can first measure an unstained sample to estimate the autofluorescence intensity. Then this intensity is subtracted from the stained sample. If every macrophage has different autofluorescence intensities. You can still estimate autofluorescence level of each macrophage through the measure of an unstained channel. This method works on our experiment of cervical cancer specimens that has large autofluorescence. If you would like to try this method and have questions, please let me know. Best Regards, Jian ************************************* Jian Ling, Ph.D. Principal Engineer Bioengineering Section Southwest Research Institute 6220 Culebra Rd. San Antonio, TX 78238 Tel. (210) 522 - 3953 Fax. (210) 684 - 6147 Email. jling@swri.org Website: www.swri.org or www.bioengineering.swri.org ************************************* -----Original Message----- From: Tracy Fischer-Smith [mailto:tlfsmith@temple.edu] Sent: Thursday, June 22, 2006 8:05 AM To: Cytometry Mailing List Subject: macrophage autofluorescence Hello, I have significant auto-fluorescence from cultured/infected/cytokine treated monocytes/macrophages that inhibits my ability to see anything by flow. I have seen some methods for quenching macrophage auto-fluorescence on the web and was hoping that if any of you have had experience with any of these methods you wouldn't mind to send me a quick response regarding what method(s) you find to be most effective. Also, would the use of a quenching dye, such as crystal violet or trypan blue affect subsequent intracellular staining? Thanks in advance! Tracy Fischer-Smith, PhD Temple University Dept. of Neuroscience Center for Neurovirology Philadelphia, PAReceived on Mon Jun 26 13:58:00 2006
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