FOR BEST RESULTS WITH MOST STAINS, USE PERFUSION SOLUTION TYPE 3.

CONSULT WITH NSA PRIOR TO PERFUSION TO DETERMINE THE MOST APPROPRIATE SOLUTIONS TO USE.

Perfusion Solutions Type 2: ‘Reagent Grade’

Phosphate Buffered Paraformaldehyde

For STANDARD and IMMUNOHISTOCHEMISTRY Stains; NOT for Disintegrative Degeneration Stain

Pre-made NSA Perfusion Fix #2 is available from EM Sciences.To order call (215) 412-8400; catalog # 1224SK.

Perfusion Fix:

Paraformaldehyde	For 1000 ml Total 40 g	For 2000 ml Total 80 g	For 4000 ml Total 160 g	Final % 4%
Sucrose	40 g	80 g	160 g	4%
Phosphate Buffer 0.2M; pH 7.2	500 ml	1000 ml	2000 ml	---

 

For 1000 ml Total:

1. Begin with 500 ml of dH20 and heat to 60°-65°C (~104° F) on a heater/ magnetic stirrer.

2. Add the sucrose.

3. 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.

4. Allow to cool to room temperature

5. Bring the total volume to 1000 ml with 0.2M Phosphate Buffer and verify the pH.

6. Filter the solution using a qualitative paper.

 

Sequence of Tissue Preparation Events with Links:

Fixation Method Fix Solutions Perfusion Instructions Fix Timing & Storage Buffers Shipping Tissue

 

Other Direct Links to Related Items:

Fixes:

• Na Cacodylate Paraformaldehyde fix solution

• Paraformaldehyde fix solution

• Technical Grade, Phosphate buffered formalin fix solution

Buffers:

• Na Cacodylate buffer solution

• Phosphate Buffered Saline

Procedures:

• Post perfusion spinal cord preparation

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