Staining w/ phalloidin and DAPI - Recommendation of staining tissue sections (Jul/17/2008 )

Hi everybody,

I just wanted to see if there were any recommendations you have on staining w/ phalloidin and DAPI for tissue section slides.
Nobody really uses it in our lab, but we have an old labstock, which I'm not sure were it's from... Since money issues... I can't really purchase another one.
I was thinking just doing a trial and error, but that usually takes a lot of time and I have no idea where to start.
All tips are welcome!

Thanks,

---

Check these links for good protocols and tips:
http://search.vadlo.com/b/q?sn=158621799&a...ction&rel=0
..

---

Hi,
We use phalloidin staining for tissue cultured cells. We use a 1:100 dilution for 1hr and then do DAPI and mount overnight in cold.
It is important not to permeabilize the cells with methanol, but triton or another method.
I hope it helps

---

Thanks,

I've been looking in to different protocols, but sometimes it's hard to take a pic and go with that. I guess, it's because you're not sure how well they work or something. But thanks for the link cell counter!
And I was planning on using Tween-20 or Triton for permeablization, but have also seen saponin. I thought that might be a bit to mild, so I think I'll just pick and just between the first 2 and see what happens.

Thanks!

---

Re. permeabilization, each tissue, species and developmental stage differs and need to be standardized in your hands. In fact, in general, any detergent can be used for any purpose if time and conc are standardized. Just do a series if you have tissue on hand. Luck!

But no Methanol!

---

we use an Alexafluor-labelled phalloidin similar as an antibody;

we also use a combination of triton-x 100 and saponin to permealize

---