Hi, I'm currently going over IHC SOPs that were written by lab members that have long ago moved on to other jobs. One inconsistency that I've come across is the use of sodium borohydride in immunofluorescence.
Our protocols state that a '0.5% sodium borohydride solution should be used to block aldehyde fluorescence if staining CNS tissue in paraffin sections' (this step is not included in our cryosection protocol). Is there a particular reason that it should only be done in CNS tissue? Seems to me that all tissues, regardless of type, should benefit from blocking aldehyde fluorescence as the problem is caused by the fixative, not the tissue type. Which leads me to the follow up question of: Our cryosection protocol should also include sodium borohydride blocking if the tissues were at any point fixed using an aldehyde fixative, correct? There's a chance I have pregnancy brain and an obvious answer is slipping past me, but my current opinion is that any and all immunofluorescence done on paraformaldehyde (our usual fixative) fixed tissue should undergo sodium borohydride blocking. Am I missing something? Thanks for any help in clarifying this! Lucie Lucie Guernsey UC San Diego lguern...@ucsd.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
