Fixation & Tissue process Protocol
Fixative
* Fresh made RNase free 4% paraformaldehyde (4%PF) used for In Situ Hybridization.
* 4% paraformaldehyde or 10% buffered formalin used for routine histology (i.e. H&E staining)
* 2% paraformaldehyde, 4% paraformaldehyde or 10% buffered formalin used Immunocytochemistry.
Fixation Procedure
The following protocols to be used for tissue sample processed for light microscopy with H&E staining, immunocytochemistry and In Situ hybridization. RNase free PBS and fixative must be used for In Situ hybridization with RNA probe.
Use 60ml yellow cap container with 30-40 ml solutions or 20ml scintillation vial with 10-15ml solutions to fix, wash and dehydrate the samples.
Well-fixed samples can be stored in same fixative at 4ûC for several days or even longer for H&E staining. In Situ hybridization sample must be dehydrated and store at -20C with 100% ethanol after 24-48 hours fixation. Generally Immunocytochemistry sample must be dehydrated and store at -20C with 100% ethanol after overnight fixation.