MBL-colour of apototic cell

[bactin]

Hi guys,
I would like to know some very basic information about TUNEL. we have a kit from MBL(Mebstain apoptosis kit direct) an insitu detection kit for programmed cell death.My question is what colour will be the apoptotic cell from the normal cell or how can i identify the apoptotic cell.This kit brochure says it uses fluorescein-dUTP to label DNA strand breaks and also advises PI for counterstaining.what colour should i expect.Thanks in advance.

[gugi]

Hi, although my kit is from "roche", fluorescein gives green signal and Propidium Iodide red. All the cells will give you a red signal, and only the apoptotic will give the green as well, so you should look for the yellow merge this way you'll know that you have an apoptotic cell. notice that green signal without a red is nor a cell!

[bactin]

Hi
Thanks for the information.Today i observed the slides under fluoroscence microscope.i am not exactly sure of which is apoptotic and which is not.I will try to post the picture as well later.Most of the cells had a red nucleus within a green cell.?apoptotic or not.

[gugi]

You have to do an overlay= merge, if the green+ the red gives you yellow, then the cell is apoptotic. Some times it's better to examine the slides under a confocal microscope in this way the PI signal (red), won't give you false green (the red is stronger than the green and some times interfere with the green)
Good luck

[bactin]

Hi,
I have been trying to post the picture(fluorscence image) but i am not able to.Is there any suggestions for me to post the picture.

[bactin]

Hi gugi
thanks for your information.i have been trying to analyse the apoptotic cell percentage using your description.But to write in the paper i would like some citation.can you please share any of it.i tried but not much fruitful as full article access is limited for me.
thank you.