抗冻缓冲液保存冰冻切片的研究

Experimantal study on the anti-freeze buffer preserving frozen sections

目的 :研究抗冻缓冲液 (anti freezebuffer,AFB)对冰冻切片的长期保存作用。方法 :4只 1 2～ 1 5月龄的雌性C5 7BL/6NIA小鼠常规灌注取脑 ,5 0 μm冰冻切片 ,将切片于 2 0℃保存于抗冻缓冲液中 ,分别于切片的当日、切片后 3个月、6个月和 1 2个月 ,行 β NADPH酶组织化学和GFAP免疫组织化学染色。结果 :于 2 0℃冰箱保存在AFB内的冰冻切片 ,至 6个月时 ,β NADPH酶组织化学显示的含一氧化氮合酶 (nitricoxidesynthase,NOS)的神经元和毛细血管与新鲜标本没有区别 ;至 1 2个月时 ,GFAP免疫组织化学染色显示的星形胶质细胞与新鲜标本没有区别。结论 :AFB可较长时间保存组织切片的抗原成份和酶活性 。

Objective To investigate whether anti freeze buffer preserves frozen sections for immunocytochemistry (ICC) and histochemistry. Methods 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℃ . At the time of sectioning (Day 0) and after 3 intervals (3, 6,and 12 months) sections through the dentate gyrus were stained with β NADPH histochemistry for nitric oxide synthase (NOS) containing neurons and capillaries, in combination with GFAP ICC for astrocytes.Results Analysis of the sections stained with β 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. Conclusion 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.