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NeuroScience Associates

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.

 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.

 

For more information:

Antigen Preserve (AP) Solution

Mounting Solutions

Perfusion Fix Solutions

Fixation Methods & Perfusion Solutions

Perfusion Protocol, Pump Calibration

Spinal Cord Post-Perfusion

Brain Extraction

Brain Hemisection

 

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