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NeuroScience Associates

Post Fixation Buffer Solutions

Tissue Storage Solution

For many “traditional” stains, the tissues may remain in the perfusion fix, for all other intended stains, please follow the instructions below. Contact NSA if you are uncertain.

Recommended time to remain in fix after perfusion (by stain type):

  • For all antibody staining: 12 hours minimum
  • For all disintegrative degeneration staining (Amino CuAg): 12-16 hours (leave brain in skull, immersed in fix, overnight before removal)
  • For all other staining: 12-16 hours
  • For immersion fixation of unperfused tissue: 12-16 hours (mouse and rat; contact NSA for larger tissue recommendation)

Storage Buffers

Following fixation, tissue should be transferred to a storage buffer (such as PBS) before 48 hours. To achieve optimal staining, the recommended guidelines of time to remain in fix and the type of buffer to which to be transferred depend on the stains that will be applied to the tissue. Contact NSA if you are uncertain.

Phosphate-Buffered Saline (PBS) is acceptable for all stains.

 500ml1000ml
Monobasic Sodium Phosphate140ml280ml
Dibasic Sodium Phosphate360ml720ml
Saline; NaCl4.5g9g

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.

 

Consult with NSA to confirm the appropriate buffer and fixation time for your study via this form or call 865.675.2245.