Microwaves and Heat in Aldehyde Fixation: Model Experiments with Bovine Serum Albumin

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°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°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 thanin vitroexperiments in which low concentrations of reagents have been used to permit subsequent spectrometry. This was confirmed by polyacrylamide gel electrophoresis of liver.