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微波固定可为光学显微镜和电子显微镜提供对组织、细胞及抗原的极佳保存效果。

Microwave fixation provides excellent preservation of tissue, cells and antigens for light and electron microscopy.

作者信息

Login G R, Dvorak A M

机构信息

Department of Pathology, Beth Israel Hospital, Boston, Massachusetts 02215.

出版信息

Histochem J. 1988 Jun-Jul;20(6-7):373-87. doi: 10.1007/BF01002732.

Abstract

It was demonstrated that microwave energy used simultaneously in combination with low concentrations of glutaraldehyde (0.05%) and formaldehyde (2.0%) rapidly preserved light microscopic histology and excellent fine structural details, as well as a variety of cytoplasmic and membrane-bound antigens. Specimen blocks up to 1 cm3 can be fixed in as brief a time as 26 ms using a specially designed microwave device (ultrafast microwave fixation method). The fast microwave fixation method, using a commercially available device, was successfully used to preserve granule-bound rat mast cell chymase which was subsequently detected by a postembedding immunogold procedure. Control of the following parameters is important to the microwave fixation method: (1) specimens with one dimension less than 1 cm; (2) irradiation temperatures lower than 50 degrees C; (3) irradiation times less than 50 s; (4) immediate replacement of the postirradiation solution with cold storage buffer; (5) fixing the specimen within 15 min after it is removed from its blood supply.

摘要

结果表明,微波能量与低浓度戊二醛(0.05%)和甲醛(2.0%)同时联合使用,能快速保存光镜组织学结构和出色的精细结构细节,以及多种细胞质和膜结合抗原。使用专门设计的微波设备(超快微波固定法),体积达1 cm³ 的标本块可在短短26毫秒内固定。利用市售设备的快速微波固定法成功保存了颗粒结合型大鼠肥大细胞糜酶,随后通过包埋后免疫金程序进行检测。控制以下参数对微波固定法很重要:(1)一维尺寸小于1 cm的标本;(2)照射温度低于50摄氏度;(3)照射时间少于50秒;(4)照射后立即用冷藏缓冲液替换溶液;(5)在标本从血供中取出后15分钟内进行固定。

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