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We are currently analysing rat peritoneal macrophages for surface=20
membrane receptors after staining with fluorescein-conjugated ligands.
"Resident" macrophages are often larger than 60 =E6, thus considerably larg=
er=20
than the 20=E6x64=E6 laser beam at excitation focus of our FACScan & FACSor=
t=20
flowcytometers. The fluorescence intensity from the cells suggestedly=20
mirrors the receptor density at the exposed part of the cell membrane,=20
but hardly reflects the total receptor pool in these cells, since=20
variation in surface area might contribute. Comparison with binding of=20
radio-iodinated ligands does not exhibit convincing correlations.
It might be common knowledge, but can someone give me a clue to=20
correction for surface area by arithmetic on FSC, or would it be far=20
reached to introduce a "topical intensity times FSC" parameter in huge cell=
s?
Yours,
Bjarne Moeller
_________________________________________________________________
Bjarne K. Moeller, MD =20
Dept. of Clin. Immunology Voice: +45 8949 5304
University Hospital of Aarhus Fax: +45 8949 6007
Skejby Sygehus =20
DK-8200 Aarhus, Denmark E-mail: bkm@biobase.dk
_________________________________________________________________
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