Lei De-Liang, Luo Xue-Gang, Peter R Mouton
Department of Anatomy and Neurobiology, Xiangya School of Medicine, Central South University, Changsha 410013, China.
Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2004 Apr;29(2):166-9.
To investigate whether anti-freeze buffer preserves frozen sections for immunocytochemistry (ICC) and histochemistry.
C57BL/6NIA female mice (n = 4) aged 12 - 15 months were used in this study. It was frozen sectioned at 50 micron and stored in anti-freeze buffer at -20 degrees C. At the time of sectioning (Day 0) and after 3 intervals (3, 6, and 12 months) sections through the dentate gyrus were stained with beta-NADPH histochemistry for nitric oxide synthase (NOS) -containing neurons and capillaries, in combination with GFAP ICC for astrocytes.
Analysis of the sections stained with beta-NADPH histochemistry revealed an equivalent level of staining for NOS-positive neurons and capillaries at both Day 0 and 6 months. Similarly, sections preserved in anti-freeze buffer and stained with GFAP ICC showed equivalent staining of astrocytes at Day 0, 6 months, and 12 months.
These findings confirm that anti-freeze buffer provides long-term preservation of antigens associated with neurons, glial cells, and blood vessels for common histological and ICC staining procedures.
研究抗冻缓冲液是否能保存用于免疫细胞化学（ICC）和组织化学的冰冻切片。
本研究使用了4只12至15月龄的C57BL/6NIA雌性小鼠。将其切成50微米的冰冻切片，并储存在-20℃的抗冻缓冲液中。在切片时（第0天）以及3个时间间隔（3、6和12个月）后，对齿状回的切片进行β-NADPH组织化学染色，以检测含一氧化氮合酶（NOS）的神经元和毛细血管，并结合胶质纤维酸性蛋白（GFAP）免疫细胞化学染色检测星形胶质细胞。
对用β-NADPH组织化学染色的切片分析显示，在第0天和6个月时，NOS阳性神经元和毛细血管的染色水平相当。同样，保存在抗冻缓冲液中并用GFAP免疫细胞化学染色的切片在第0天、6个月和12个月时显示星形胶质细胞的染色相当。
这些发现证实，抗冻缓冲液可为常见的组织学和免疫细胞化学染色程序长期保存与神经元、神经胶质细胞和血管相关的抗原。
