Hi - It is difficult to diagnose these kinds of problems remotely without more information (instrument, conditions of running), but just from looking at the patterns - sample 1 not only has low FSc for granulocytes, but lower signal levels and smearing of the populations in R1 and R2 - I would guess that there is an alignment issue. If the cytometer has fixed alignment and sample 2 was run shortly after sample 1, then I would suspect a contaminant in sample 1 that is killing the cells. Marty Bigos, Director Stanford Shared FACS Facility mail: Stanford Shared FACS Facility Beckman Center B015 Stanford CA 94305-5318 Phones: (office) 650-725-8463 (mobile) 415-845-8450 fax: 650-725-8564 On Jun 28, 2008, at 2:19 PM, Abdul Khan wrote: > Dear Flowers, > > I am a PhD student and doing flow cytometry on peripheral blood > from healthy women. So far I have done quite a few samples and all > of them > are quite consistent in terms of results, for example CD3, CD4, CD8, > CD14, > CD66b expression. The attached FSC/SSC plot (FSC-SSC -1 file) looks > very > unusual to me as you can see most of the granulocytes are having low > FSC and > I am wondering if anyone have come across with this earlier and would > appreciate any help/comments. First I thought that there might be some > problem with the machine, so I did an experiment on my own blood > (FSC-SSC -2 > file) just to make sure that FACS machine is working fine and indeed > I found > the same reproducible result. > > I sincerely hope that someone here will be able to help me out in > understanding what could be wrong here! Thank you all. > > > Best wishes > > Abdul Khan > PhD Student > University of Glasgow > This attachment - 'FSC-SSC -1 unstained 260608 .JPG' - 22.31 KBytes > - can be viewed at > http://www.cyto.purdue.edu/MD-parts/286724e98a43dc7207e01fca3ffcf8e3abac4964.JPG > > This attachment - 'FSC-SSC - 2 unstained 270608.JPG' - 22.07 KBytes > - can be viewed at > http://www.cyto.purdue.edu/MD-parts/0cc2a65dbcdb1e3b6a979e0fe90bbb1713b62d92.JPG >Received on Wed Jul 9 13:58:00 2008
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