Post-Fixation Buffer Solutions, Storage and Timing
Recommended time to remain in fix after perfusion is 12-16 hours.
- Antibody staining: for formalin sensitive antibody staining 4 hours post perfusion is sufficient
- Amino CuAg (degeneration staining): wait at least 24 hours before extracting brain to allow brains to fully harden, preventing the induction of artifacts during staining
Following fixation, tissue should be transferred to a storage buffer, Phosphate-Buffered Saline (PBS) which is acceptable for all stains.
Monobasic Sodium Phosphate 140ml 280ml
Dibasic Sodium Phosphate 360ml 720ml
Saline; NaCl 4.5g 9g
Concentrated PBS is also commercially available from Sigma-Aldrich, Catalog# 79383-1L.
Phosphate-Buffered Saline (PBS) with 0.01% sodium azide should be used for tissue stored over three (3) weeks. Storage for this time period should take place in a refrigerator (4 degrees C).
Also see: Use of Cryroprotectant to Maintain Longterm Peptide Immunoreactivity and Tissue Morphology by Hoffman and Le.
For more information:
Consult with NSA to confirm the appropriate buffer and fixation time for your study via this form or call 865.675.2245.