中性红是一种弱碱性pH指示剂,变色范围在pH6.4~80之间 (由红变黄)。分子式为 C₅H₁₆N₄·HCl,分子量为288.78。在中性或微碱性环境中,植物的活细胞能大量吸收中性红并向液泡中排泌。中性红染色液 (0.5%) 是一种组织或细胞染色时常用的可以把细胞核染成红色的染色液,常用于组织或细胞染色中细胞核的红色复染。
操作步骤(仅供参考):
1、样品处理
a)对于石蜡切片:
二甲苯中脱蜡5~10min。
换用新鲜的二甲苯,再脱蜡5~10min。无水乙醇5min。90%乙醇2min。70%乙醇2min。蒸馏水2min。
b)对于冰冻切片:蒸馏水2min。
c)对于培养细胞:用4%多聚甲醛固定10min 以上。蒸馏水洗涤2min。换用新鲜的蒸馏水,再洗涤2min。
2、中性红染色
a)对于上述处理好的样品,用Neutral Red stain (0.5%) 染色。
b)用蒸馏水或自来水充分洗涤。
c)进行观察和拍照。
注意事项:
1、如果需要脱水、透明和封片处理,还需自备二甲苯,中性树胶或其它封片剂。如果样品是石蜡切片,需自备70%和90%乙醇,无水乙醇以及二甲苯。
2、首次使用本试剂时建议先取1~2个样品做预实验。
3、中性红染色液时,可以根据染色结果和要求调整时间。
4、为了您的安全和健康,请穿实验服并戴一次性手套操作。
5、本产品仅用于科研,严禁他用。
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.