Login G R, Dvorak A M
Department of Pathology, Harvard School of Dental Medicine, Boston, MA.
Prog Histochem Cytochem. 1994;27(4):1-127.
Microwave fixation methods are important because excellent preservation of both cell structure and antigenicity can be attained several orders of magnitude faster than by routine chemical fixation methods. Fast and ultrafast microwave fixation have yielded significant logistic advantages over another fast fixation approach-rapid freezing at liquid helium temperatures. For example, specimens used for microwave fixation can be as large as 1 cm3 and cells can remain in suspension. We review in detail both qualitative and quantitative morphologic results obtained by using microwave fixation in sample preparation. We provide tables of biological molecules that are preserved in a variety of human and animal tissues by various microwave fixation methods for histochemistry, immunohistochemistry, cytochemistry, immunocytochemistry, and affinity labelling studies. Limitations of large cavity (e.g., household) microwave ovens often result in irreproducible fixation results. We present calibration and standardization protocols for microwave fixation in large cavity microwave ovens that emphasize a) localization of oven hot spots (i.e., high power) using a neon bulb array, b) magnetron warm-up, c) the use of a water load, d) the use of an agar-saline-Giemsa model to predict the uniformity of irradiation in small samples, e) the use of specimen containers with one dimension less than 1.5 cm, and f) fast specimen handling to prevent conductive heating artifacts after irradiation. Although microwave ovens are commonplace their unique applications in the laboratory environment require special safety considerations, which are reviewed. Advances in microwave technology are providing new means to study the structure-function relationships of cellular and biochemical activities.
微波固定方法很重要,因为与常规化学固定方法相比,它能在快几个数量级的时间内实现细胞结构和抗原性的出色保存。快速和超快微波固定与另一种快速固定方法——在液氦温度下快速冷冻相比,具有显著的后勤优势。例如,用于微波固定的标本可以大到1立方厘米,细胞可以保持悬浮状态。我们详细回顾了在样品制备中使用微波固定获得的定性和定量形态学结果。我们提供了通过各种微波固定方法保存在多种人类和动物组织中的生物分子表格,用于组织化学、免疫组织化学、细胞化学、免疫细胞化学和亲和标记研究。大型腔体(如家用)微波炉的局限性常常导致不可重复的固定结果。我们提出了大型腔体微波炉中微波固定的校准和标准化方案,重点强调:a)使用霓虹灯灯泡阵列定位微波炉热点(即高功率区域);b)磁控管预热;c)使用水负载;d)使用琼脂-盐水-吉姆萨模型预测小样品中辐射的均匀性;e)使用一维尺寸小于1.5厘米的标本容器;f)快速处理标本以防止辐射后产生传导加热伪像。尽管微波炉很常见,但它们在实验室环境中的独特应用需要特殊的安全考虑,本文对此进行了综述。微波技术的进步为研究细胞和生化活动的结构-功能关系提供了新手段。