studying apoptosis - involves flourscence microscopy (Sep/11/2006 )

Hi All,
I am planning to perform flourscence microscopy to study apoptosis, that will involve studying condensation of chromatin in side the cell nucleus.
So far, I have read about ethidium bromide and acridine orange being used as dyes for this purpose.
Are there other alternative dyes/reagents/methods that can be used? And are there any problems with using the dyes I have mentioned?
Thanks,
Penny

u could try DAPI- stains nucleus, u would c condensed chromatin in apoptotic cells.

I've heard that annexin V is also used for apoptosis labeling (it can be coupled to a fluorescent dye).

Hi All,
I am planning to perform flourscence microscopy to study apoptosis, that will involve studying condensation of chromatin in side the cell nucleus.
So far, I have read about ethidium bromide and acridine orange being used as dyes for this purpose.
Are there other alternative dyes/reagents/methods that can be used? And are there any problems with using the dyes I have mentioned?
Thanks,
Penny

Thank you everyone for your positive suggestions,
Penny

I also like to use propidium iodide

Hi,
I finally deicded on using acridine orange/eithidium bromide and performing fluorescence microscopy. However, my cells are not adherant but in suspension. Also, I want to make a cell count rather than a qualitative analysis, so is there a special kind of slide available that can be used for counting cells in this situation? I heard of 6d-plates but that seems like an expensive option and probably is for qualitative analysis, not quantitative.

ALso, I read about paraformaldehyde fixation....is it necessary...all I know is that you can actually preserve cells in that state for a week ...... but then is it recommended for use on live cells, I mean isnt it supposed to be toxic to cells and actually kill them? As far as I have understood that cells do die but then you are still able to distinguish between cells as healthy, apoptotic, and necrotic cells. I am not sure but am trying to understand it. Please point out mistakes in my understanding of the concept, if there be. And suggest improvements...in the methodology discussed above.
Thank you all,
Penny