[Experimantal study on the anti-freeze buffer preserving frozen sections].
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.


CONCLUSIONS
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.
Lei
De-Liang
DL
Department of Anatomy and Neurobiology, Xiangya School of Medicine, Central South University, Changsha 410013, China.
Luo
Xue-Gang
XG
Peter
R Mouton
RM
D000818:Animals; D001921:Brain / Q000033:anatomy & histology*; D002021:Buffers; D005260:Female; D005629:Frozen Sections / Q000379:methods*; D051379:Mice; D008810:Mice, Inbred C57BL; D009252:NADPH Dehydrogenase / Q000378:metabolism; D052248:Nitric Oxide Synthase Type I / Q000378:metabolism; D013194:Staining and Labeling; D013997:Time Factors
D002021:Buffers; D052248:Nitric Oxide Synthase Type I; D009252:NADPH Dehydrogenase