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醛类固定中的微波与加热:牛血清白蛋白的模型实验

Microwaves and heat in aldehyde fixation: model experiments with bovine serum albumin.

作者信息

Hopwood D

机构信息

Pathology Department, Ninewells Hospital and Medical School, Dundee, Scotland.

出版信息

Methods. 1998 Jun;15(2):119-22. doi: 10.1006/meth.1998.0614.

Abstract

Most model experiments concerning tissue fixation have used low concentrations of fixatives and proteins. Here, high concentrations (up to 32%) of bovine serum albumin (BSA) were reacted with formaldehyde (1-20%) and glutaraldehyde (0.5-4%). Gels were formed between 16% BSA and 10-20% formaldehyde at room temperature, but not with percentages of formaldehyde lower than 4%. Microwave application or heating in a water bath to 50 degrees C gave a gel from 1 to 20% formaldehyde. Sixteen percent BSA also gave a gel with glutaraldehyde from 0.5 to 4%. Cone and plate viscometry showed rapidly increasing viscosity at 4% formaldehyde and 16% BSA at room temperature. At 50 degrees C, gels formed at a low concentration of formaldehyde. Tissue fixation in which the local concentrations of protein may be in excess of 30% is probably more complete than in vitro experiments in which low concentrations of reagents have been used to permit subsequent spectrometry. This was confirmed by polyacrylamide gel electrophoresis of liver.

摘要

大多数关于组织固定的模型实验都使用了低浓度的固定剂和蛋白质。在此,高浓度(高达32%)的牛血清白蛋白(BSA)与甲醛(1%-20%)和戊二醛(0.5%-4%)发生反应。在室温下,16%的BSA与10%-20%的甲醛之间形成了凝胶,但甲醛浓度低于4%时则不会形成凝胶。通过微波处理或在水浴中加热至50摄氏度,1%-20%的甲醛均可形成凝胶。16%的BSA与0.5%-4%的戊二醛也可形成凝胶。锥板粘度测定显示,在室温下,当甲醛浓度为4%且BSA浓度为16%时,粘度迅速增加。在50摄氏度时,低浓度的甲醛即可形成凝胶。与使用低浓度试剂以便后续进行光谱分析的体外实验相比,局部蛋白质浓度可能超过30%的组织固定可能更彻底。肝脏的聚丙烯酰胺凝胶电泳证实了这一点。

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