1 reply to this topic

[juliai]

What is a protocol of Wright staining for apoptosis analysis in K562 cells?

[KDHughes]

Try putting a drop of your cell suspension onto a clean slide and letting it dry.  The cells shouldn't need any further fixation because of the alcohol in the stain.  Cover the dried cells with 10-15 drops of Wright's stain and allow to sit at room temperature for about five minutes; add an equal volume of Wright's buffer and let it sit for another five minutes.  After that rinse the slide in distilled water and allow to air dry.

If you have access to a fluorescent microscope, I've had some luck staining cells (both adherent and in suspension) with acridine orange and ethidium bromide.  Using these two together, you can differentiate between four classes of cells: live non-apoptotic cells, live apoptotic, necrotic (dead non-apoptotic), and dead apoptotic.