immunos and pigmentation

[Declan W Ali]

My name is Declan Ali and research in my lab focuses on the 
physiology of synaptic development. We use zebrafish as our 
organism of study and have recently taken up performing 
immunohistochemistry on wholemounts of embryos and larvae. We 
are having some trouble with background and transparency in older 
animals (3-5 days), particularly with the presence of pigmentation. 
We rear larvae in 0.03% PTU which retards pigmentation but I am 
not happy with the strength of the specific staining in PTU, and 
was wondering if you knew any other method of clearing pigments. 
Since our specific staing is a bit weak it doesn't help that the 
background and pigmentation is high in older animals. We use 
FITC or Cy3 coupled to our secondary and clear the wholemounts 
in glycerol.  

Can someone help us?

Sincerely
Declan

Declan W Ali, PhD
Assistant Professor of Zoology
University of Alberta
Department of Biological Sciences
CW-405 Biological Sciences Bldg.
Edmonton, Alberta
Canada, T6G 2E9
Tel: (780) 492-6094
Fax: (780) 492-9234
Email: declan.ali at ualberta.ca