john d

I'm not really into their music and I know absolutely nothing about the band, but it just seems weird that a lot of people hate them so much. What makes them so hatable, other than saying "their music sucks"? I Wanna be a Rock Star sounds okay, imho.

I'm staining for a cell surface receptor on cultured cells, to be analyzed with FACS. I need to have the data ready in a month, any information would be good!

And how do I decide when to use ANOVA or t test? Are they interchangable?

I've seen some weird results, so......

After some paper-reading, I realized there are two ways to quantify TUNEL: either by flow cytometry, or using chamber slides or cytospin to adhere cells to a glass slide, performing TUNEL with an in-situ kit, and then counting by direct microscopy.

When I presented this discrepency to a senior lab member, he just gave me a cold stare and asked me which was more accurate: counting 400 cells or 10,000? However, I think there must be some reason why so many papers use the glass slide method !

Does it have something to do with the adhesiveness of cells (all the papers using glass slides use adhesive cells)? After all that failure and disappointment I went through with annexinV/PI, I don't want to make the same mistake again!

And another question: If I'm doing TUNEL via flow, is single staining enough or should I counter-stain it with PI or something? I'm not sure what the function of counter-staining in this situation actually is.... Anyways I'm a newbie in this apoptosis thing (my prof asked me to do this two months ago), any help at all would be greatly appreciated!

Hi, I want to detect DNA fragmentation using TUNEL (Terminal Transferase dUTP Nick End Labeling), but we don't have a flourescent microscope. I heard that it's possible to use a flow cytometer for quantification, are there any good protocols for doing that? Also, do I need to do a double stain like in AnnexinV-PI, or will single staining with be enough?

Uh, just testing......