Perfusion Fix Solutions
High quality perfusion fix solutions yield high quality results. Commercially available formaldehyde solutions are NOT fresh and usually have added ingredients to stabilize the solution for long shelf life. This should be considered a “technical grade” perfusion fix solution. Such solutions should only be used for standard, routine, classical stains. However, even these stains may be optimized by having a freshly prepared perfusion solution using reagent grade components.
Preferred Perfusion Fix Solution
Phosphate-Buffered Paraformaldehyde:
Order from EM Sciences, call (215) 412-8400; catalog # 1224SK.
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Prepare in the lab:
For 1000 ml Total For 2000 ml Total For 4000 ml Total Final %
Paraformaldehyde 40 g 80 g 160 g 4%
Phosphate Buffer
0.2M; pH 7.2500 ml 1000 ml 2000 ml --
Sucrose
Note: Not to be used for the purpose of cryoprotection40 g 80 g 160 g 4%
For 1000 ml Total:
- Begin with 500 ml of dH20 and heat to 60°-65°C (~104° F) on a heater/ magnetic stirrer.
- Add the sucrose. Please note: Sucrose is optional and this solution is not to be used for cryoprotection.
- Create a deep vortex using a magnetic stirrer bar. Gradually sprinkle the finely powdered paraformaldehyde onto the vortex surface. The solution will be quite milky.
- Allow to cool to room temperature
- Bring the total volume to 1000 ml with 0.2M Phosphate Buffer and verify the pH.
- Filter the solution using a qualitative paper.
For more information:
Antigen Preserve (AP) Solution
Post-Fixative (Storage) Buffer Solutions
Contact Technical Support at NSA Labs via this form or call 865.675.2245 for shipping and preparation questions.
