Processing of Tissue Specimens
Processing of Tissue Specimens
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DOI：
10.1007/978-1-59745-324-0_13
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发表时间：
2010-01-01
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影响因子：
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通讯作者：
Bratthauer, Gary L.
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作者：
Bratthauer, Gary L.
为了用抗体测试组织标本，首先必须将它们保存在固定剂中，包埋在石蜡中，并在玻璃显微镜载玻片上切成非常薄的切片。任何一片组织，在切除后，必须立即放入足够量的固定剂中。固定剂不同，但标准的是10%缓冲福尔马林。在最佳固定时间（对于福尔马林，约16小时）后，必须将样品包埋在石蜡中并在切片机上切片。然后，将带正电荷的载玻片（涂层或商业制备的）置于石蜡切片中，准备进行各种预处理步骤。首先，石蜡必须通过一系列的再水化步骤用水代替。然后，根据待测抗原，切片可以用酶进行蛋白水解消化或在低或高pH溶液中进行热处理。之后，内源性过氧化物酶或氧化性化合物可以在过氧化氢溶液中淬灭。在正常血清溶液中孵育后，切片准备好用抗体进行测试，溶液阻断任何可用的带电位点。
In order to test tissue specimens with antibody, they first have to be preserved in fixative, embedded in paraffin, and sectioned very thinly onto glass microscope slides. Any piece Of tissue, immediately after excision, must be placed into an adequate volume of fixative. Fixatives vary, but the standard one is 10% buffered formalin. After an optimum fixation time (for formalin, about 16 h), the sample must be embedded in paraffin and sectioned oil a microtome. Paraffin-ern bedded sections placed oil positively charged slides (either coated or commercially prepared) arc then ready for various pretreatment steps. First, the paraffin Must be replaced with water through a series of rehydration steps. Then, depending on the antigen to be tested, the section can be proteolytically digested with enzymes or heat-treated in low or high pH solutions. Following that, the endogenous peroxidase enzyme or oxidative Compounds can be quenched in a hydrogen peroxide solution. The sections arc then ready to be tested with antibody after an incubation in a normal serum Solution blocks any available charged sites.