Quantification of apoptosis - (Sep/14/2006 )

Hi
can anyone please let me know a good method of quantifying apoptosis
May be any ELISA which is reproduceable , I am having some problems with flow cytometry
Thanks
sweta

Hi Shweta,
You not here told which types of problems you facing by using FACS because i thinks its
a good technique for apoptosis detection. But you can use some other techniques like Flourescence microscopy for tuneel assay, early apoptosis detection and late apoptosis by DAPI and some other dye staining.
You also can use specific substrates for caspases estimation.
If you has problems with FACS you can disscuse with me without any hesitation.
all the best and good luck.
awadh

we routinely use an anti-histone-DNS ELISA kit from Roche detecting oligonucleosomes; there exist various other ELISA f.i. to detect activity of various caspases or their cleavage products

Hi,
I am planning to use fluoresecnce microscopy to do a cell count of healthy, apoptotic, necrotic cells.
But is there a way I can make an accurate cell count each time (in terms of cells/ml). An equivalent of haemocytometer, used for trypan blue staining.
Or else probably someone can suggest an alternative,
Thanks all!