McElroy H H, Shih M S, Parfitt A M
Bone and Mineral Research Laboratory, Henry Ford Hospital, Detroit, Michigan 48202.
Biotech Histochem. 1993 Jan;68(1):50-5. doi: 10.3109/10520299309105578.
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% carboxymethylcellulose 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 cryocompound 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.
我们描述了一种用于快速制作和保存新鲜冷冻骨活检样本的方法，这些样本可用于多种免疫组织化学技术。切除后5分钟内，将组织放入冷的5%聚乙烯醇中，置于手工制作的铝箔包埋模具中，模具中填充3%羧甲基纤维素，然后浸入-70℃的无水乙醇/干冰浆液中冷冻。用低温化合物将组织块固定在标本短轴上，并安装在-32℃的低温恒温器中，该恒温器的碳化钨D型刀保持在-70℃。自动控制装置设置为慢速切割速度，并调整“切片窗口”以适应活检样本的大小。改变刀角、厚度测量仪和防卷条以切出完整的切片。用浸有聚乙烯吡咯烷酮（PVP）的罗斯镜头纸条将组织块表面平滑地“覆盖”。切下一片切片并放置在依次编号、酸洗、双浸铬明矾明胶包被的载玻片上；用吸水纸“压印”有助于切片黏附。切片保存在-20℃或室温下的干燥器中。经过短暂固定，然后去除水溶性PVP和镜头纸，即可得到适合进一步分析的新鲜冷冻骨切片。
