Neutral Red is a weakly basic pH indicator with a color change range of pH 6.4–8.0 (turning from red to yellow). Its molecular formula is C₅H₁₆N₄·HCl and the molecular weight is 288.78. In a neutral or slightly alkaline environment, living plant cells can absorb a large amount of Neutral Red and secrete it into vacuoles.
Neutral Red Stain Solution (0.5%) is a commonly used stain for tissue or cell staining that colors cell nuclei red, and it is frequently applied for the red counterstaining of cell nuclei in tissue or cell staining procedures.
Operating Procedures (For Reference Only):
1. Sample Preparation
a) For paraffin sections:
Dewax in xylene for 5–10 minutes.
Replace with fresh xylene and dewax for another 5–10 minutes. Rinse in absolute ethanol for 5 minutes, 90% ethanol for 2 minutes, 70% ethanol for 2 minutes, and distilled water for 2 minutes.
b) For frozen sections: Rinse in distilled water for 2 minutes.
c) For cultured cells: Fix with 4% paraformaldehyde for more than 10 minutes. Rinse in distilled water for 2 minutes, replace with fresh distilled water, and rinse for another 2 minutes.
2. Neutral Red Staining
a) Stain the above-prepared samples with 0.5% Neutral Red Stain.
b) Rinse thoroughly with distilled water or tap water.
c) Proceed with observation and photography.
Precautions:
1. Xylene, neutral balsam or other mounting media shall be prepared by the user if dehydration, clearing and mounting are required. For paraffin section samples, 70% ethanol, 90% ethanol, absolute ethanol and xylene shall be prepared by the user.
2. A pre-experiment with 1–2 samples is recommended when using this reagent for the first time.
3. The staining time of Neutral Red Staining Solution can be adjusted according to the staining results and experimental requirements.
4. Wear a lab coat and disposable gloves during operation for your safety and health.
5. This product is for research use only and shall not be used for any other purposes.
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