In the main, we would agree with your comments, and welcome your input. I note your comments to the list in January 1999 on practically the same subject. One question though - we gate on the area of our FS v SS plot where we expect granulocytes to be, and surely one would not expect bare nuclei to lie within that gate? Initially, we had been a little concerned about this possibility ourselves, but after further thought on the matter, we feel that this is unlikely. What do you think? We may try to get access to a sorter, also, tho' we don't have direct access at present. sorting this sub-population, and looking at some slides seems like a very good idea! Thanks a lot for your advice. Best wishes, Martin Kelly On Wed, 9 Feb 2000 17:26:40 -0500 Michael Ormerod <Michael_Ormerod@compuserve.com> wrote: > Bare nuclei will stain PI+ve, Annexin V-ve. If you have access to s sorter, > I would sort a few onto a microcope slide and have a look at them. > Otherwise if look at some samples under a fluorescence microscope, you can > check if there are any bare nuclei. > > Cells in an advanced stage of necrosis will have a lower DNA content > (because the DNA will be heavily degraded. I would expect to find them in > the Annexin +ve populatin with an intermediate PI staining. > > Michael Ormerod > > Message text written by Martin Kelly > >Dear friends, > we would like to ask some advice r.e. the interpretation of some > work we have been doing on induced sputum and BAL from normal > subjects and COPD patients. We have been using 'flow' to look at > the granulocytes, and examine the AV and PI staining in the > events which we have gated on, which we feel are in the region > in which one would expect to find granulocytes. Cells coming up > AV+PI- we interpret as apoptotic; AV+PI+ as necrotic. This would > seem to be conventionally how it is desribed in papers we have > looked at. However, we were not sure how one might interpret > cells which are AV-PI+? Are these just cells in an advanced > state of necrosis? Is there another interpretation? > We value any advice. > With best wishes. yours sincerely, > > Martin Kelly< > ---------------------- Martin Kelly Department of Clinical Biochemistry, Institute of Clinical Science, Grosvenor Road, BELFAST. BT12 6BJ Tel No: (01232) 263267
This archive was generated by hypermail 2b29 : Sat Mar 10 2001 - 19:31:06 EST