[Histonet] CD4/CD25/TGFB Antibodies; Filter choices for Leica

[Juan Solon]

Dear All,

As part of a study on mucosal immunity in The Gambia, W. Africa, I plan to double stain antral and duodenal specimens for CD4 and CD25 & CD8 and CD25.  Also, we would attempt to triple stain for TGF beta on these subsets.  We have access to a Leica DMRXE although the filter cubes are all currently long pass suppression filters . I have both formalin-fixed and frozen sections of the antrum and the duodenum.  

I would like to ask for advise on several points:

1.	Can anyone share their experience on antibody combinations known to work for CD4-25, CD8-25 and triple staining for CD4-CD25-TGFB either with vectastain elite (peroxidase) or immunofluorescence.  I have compiled a list of CD4, CD8, CD25, TGFB antibodies from Dako, Ancell, Novocastra, BD, and Serotec, but before purchasing any of these, I thought it would be wise to ask advise from the Histonet community.

2.	I would probably need to acquire different filter cubes to do simultaneous imaging of fluorescence.  There are several suppliers I have encountered on the net (Omega Filters and Chroma Technology) * does anyone have experience on using filters from these companies on Leica microscopes (specifically the DMRXE).  Any problems encountered with using third-party filters with Leica microscopes?

Thanks.
Juan


Dr. Juan Antonio Solon
MRC Keneba Field Station
MRC Laboratories Gambia
Atlantic Road, Fajara
PO Box 273, Banjul
The Gambia
Email :  juan.solon <@t> lshtm.ac.uk
Tel No:  (++220) 541021
Fax No: (++220) 541022 / (++220) 496513