[Histonet] RE: Fluorescence staining

[C.M. van der Loos]

   Dear Eva,

   To my knowledge most of the traditional fluorochromes like FITC,
   rhodamine, doesn't stand dehydration at all. So in your case it fully
   depends on what type of fluorochrome you end up with after the TSA
   procedure.

   If you end up with FITC, the use of VectaShield
   ([1]www.vectorlabs.com) for mounting is highly recommended for
   the prevention of photobleaching.

   Chris van der Loos, PhD
   Dept. of Pathology
   Academical Medical Center M2-230
   Meibergdreef 9
   NL-1105 AZ Amsterdam
   The Netherlands

   phone:  +31 20 5665631
   fax:    +31 20 6960389
   e-mail: [2]c.m.vanderloos <@t> amc.uva.nl


   ----- Original Message -----
                                  From 
   Eva C Anderson <eca9 <@t> georgetown.edu>
                                  Date 
   Tue, 25 Jan 2005 11:10:41 -0500
                                    To 
   histonet <@t> lists.utsouthwestern.edu
                               Subject 
   [Histonet] Fluorescence staining

   Hi,
   We  are looking into trying some Fluorescence staining. We want to try
   a  TSA  Fluorescence  System  from  PerkinElmer. The IHC protocol does
   however leave me with some questions.
   We  always  dehydrate our samples after staining. The protocol however
   doesn't mention this. It only says that the samples can be
   counterstained and mounted after incubation with the Fluorophore
   Tyramide. Should we dehydrate and is there a particular mounting media
   we  should  use to prolong the time that fluorescence can be detected?
   Any help would be greatly appreciated.
   Thanks in advance for all your help,
   Eva
   Georgetown University
   Research Assistant

References

   1. http://www.vectorlabs.com/
   2. mailto:c.m.vanderloos <@t> amc.uva.nl