HOW TRYPHAN BLUE STAINING WORKS? - (Jul/05/2006 )

Hi,

I am working with cell lysis for protein extraction. I want to study if there is cell membrane rupture due to the protocol , i am using now.
i thought i could try with trypan blue and phase contrast microscopy.Though they say itis not sensitive but it qualitatively works. And also,i want to know how the cell is breaking open.

can this be effectively done through trypan blue and phase contrast microscopy??
please post your suggestions..

I want to know how trypan blue gets the dead cells.. The mechanism i mean .... by knowing this i can analyse my protocol...


thanking you,
Annie D Souza

hi,
living cell's plasma membrane is semi permeable in nature. so they do not allow this dye to enter inside the cells as they consider it as foriegn molecule.
where as dead cells does not have this control system, dye enters inside the cell and stain them.
at the end you count living cells which u can observe in bright color. hence forth this technique is called DYE EXCLUSION method.

i dont think that u can study membrane rupture process with this method.

bye
gud luk
payeli