Neutral red
for Nuclei

Solutions

Staining solution
Neutral red	1	g
Acetic acid, glacial	1	drop
Distilled water	100	mL

To avoid clumping, dust the dye onto the surface of the water while

shaking it. Leave overnight, add the acid, and filter.

Tissue sample
5µ paraffin sections of neutral buffered formalin fixed tissue are suitable. Other fixatives are likely to be satisfactory.

Method

1. Complete the primary stain.
2. Place into the solution for 1 - 3 minutes.
3. Rinse well with water.
4. Dehydrate with ethanol and clear with xylene.

Expected results

• Nuclei  –  red
• Other tissues  –  as the primary stain

Notes

1. The small amount of acetic acid slightly acidifies the solution and reduces background staining, making the solution more selectively nuclear. Do not add more than specified.
2. The ethanol used for dehydration removes some of the stain. This permits a small degree of control.
3. If necessary, staining can be removed with acid alcohol. However, check that the primary stain will not be affected before doing so.

 

Reference
There is no specific reference for this application.