Hopwood D
Pathology Department, Ninewells Hospital and Medical School, Dundee, Scotland.
Methods. 1998 Jun;15(2):119-22. doi: 10.1006/meth.1998.0614.
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.
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。