Producing Frozen Sections of Calcified Bone
作者:
McElroyH. H.,
ParfittA. M.,
ShihM. S.,
期刊:
Biotechnic&Histochemistry
(Taylor Available online 1993)
卷期:
Volume 68,
issue 1
页码: 50-55
ISSN:1052-0295
年代: 1993
DOI:10.3109/10520299309105578
出版商: Taylor&Francis
关键词: Cryosections;undecalcified bone;polyvinylpyrrolidone (PVP). carboxymethylcellu-lose (CMC);polyvinyl alcohol. (PVA);OCT. lens paper
数据来源: Taylor
摘要:
We describe a procedure for the rapid production and maintenance of fresh frozen bone biopsies which can be used for a variety of immunohistochemical techniques. Within 5 min of excision. tissue is placed in cold 5% polyvinyl alcohol, surrounded with 3% carboxymethylcel-lulose in a hand made aluminum foil embedding mold and frozen by immersion in an absolute ethanol/dry ice slurry at -70 C. The tissue block is attached to the specimen stub with cryocom-pound and installed in a -32 C cryostat whose tungsten carbide D profile knife is maintained at -70 C. Automatic controls are set at a slow cutting speed and the“sectioning window”is adjusted to fit the biopsy size. Knife angle, thickness gauge and antiroll bar are changed to produce a complete section. The block face is smoothly“papered”with a polyvinylpyrrolidone (PVP) impregnated Ross lens paper strip. A single section is cut and positioned on a sequentially numbered, acid cleaned, double dipped chrome-alum gelatin coated slide: adhesion is aided by“press-blotting”with bibulous paper. Sections are stored at -20 C or in a desiccator at room temperature. A brief fixation followed by removal of the water soluble PVP and lens paper generates fresh frozen bone sections suitable for further analysis.
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