Dear FCM users, I am enquiring if anyone has assessed endocytosis by FCM. My acquisition results by FCM have shown that all the cells were positive independent of the incubation temperature (on ice or 37C). These endocytic probes must still be sticking to the outside of the cell surface causing background. Treatment with trypan blue to quench the endocytic markers from the outside of the cell surface has not seemed to help the situation. Endocytic markers used by FCM - Molecular Probes BODIPY Zymosan (Z-2844) - measures by mannose-receptor BODIPY FITC-Dextran (D-1821 or D-1844) - measures by mannose-receptor Lucifer Yellow (L-453) - meaures fluid-phase macropinocytosis LDL - (L-3483) - measures through LDL-receptor Does anyone have any suggestions or established protocols to remedy the situation? As of yet, I am not a registered user of this particular FCM mailing list. Could someone please instruct me how to register since I will not be able to retrive the following comments. Thank you for your time and assistance . I hope the information obtained helps others as well! -- Carlos P. Carrasco, PhD student Department of Immunology Institute of Virology and Immunoprophylaxis (IVI) Sensemattstrasse 293 3147 Mittelhaeusern Switzerland Tel.: +41 +31 84 89 245 Fax.: +41 +31 84 89 222 e-mail: carlos.carrasco@ivi.admin.ch
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