ABSTRACT
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 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. RESULTS: 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. 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.
