[Histonet] VIP and residue problem with Alexa

[Leila Etemadi]

Hi every body,

I would like to get your keen ideas about two issues I am facing now in my immunohistochem. procedure:

1- I am using goat anti rabbit Alexa 594 (red) as secondary antibody for detecting C-Fos in rat nervous system. When I am looking at the slides, it seems always I have some residues of my
Alexa remained on the surface all over the section. I've added one extra quick wash in every washing steps ( Using PBS+TX100). But still they are exist. Is any body has suggestion to improve the quality of the picture and get ride of these residues?


2- I need some suggestions for detecting VIP in spinal cord and DRG. In first place it is not easy to count the cells which are expressed VIP. Do you think I should go for ratio of the expression?,
Second satellite cells in DRG make it so hard to determine VIP expression is belong to them or neurones!, I was thinking to use HuC/HuD to facilitate procedure, Is there any other suggestions out there?


 By the way I will be so appreciate if any one address me a nice picture of VIP expression in DRG. I would like to see how it look like in intact animal ( preferably rat). Unfortunately, most of the articles have black and white/ or poor pictures.

Thanks for your valuable time,

Leila :-)





Leila Etemadi
M.Sc., Ph.D Candidate
Neuronano Research Center (NRC)
Lund University, BMC F10
Sweden

Tel: +46-46-2221503
E-mail: Leila.Etemadi <@t> med.lu.se<mailto:Leila.Etemadi <@t> med.lu.se>