hi, i'm at novice at cytometry. i would like to try to isolate monocytes from Gaucher disease patient's peripheral blood that are showing some of the characteristics of full blown Gaucher cells as found in the spleen. i've found the best ab to use is PCNA. i then need to demonstrate that the monocytes i will separate that are cd14+/PCNA+ have characteristics of mature Gaucher cells, under EM/light microscopy. This characteristic happens to be these inclusion bodies found in gaucher cells. They are a minimum of 0.3 microns in diameter and consist of complex tubular mixtures of glycoproteins and glycolipids. the questions then is, what effect will the permeabilizing i need to do to use PCNA have on these structures? I'm planning to use the cytofix/cytoperm kit (with formaldehyde and saponin). keep in mind that these things are not free floating fats in the cytoplasm. Is the permeabilizing likely to destroy them? is there a 'best' way to permeabilize in this case? thanks alot. --- Seth L. Ness M.D., Ph.D Fellow in Human Genetics Department of Human Genetics Mount Sinai School of Medicine Phone:212-241-6947 Fax:212-860-3316 sln8@columbia.edu Ness Gadol Haya Sham
This archive was generated by hypermail 2b29 : Sun Jan 05 2003 - 19:26:12 EST