A comparison of the effects of three widely used glutaraldehyde fixatives on cellular volume and structure. A TEM, SEM, Volumetric and Cytochemical Study

Abstract

The effects of three widely used glutaraldehyde-based fixatives on cellular volume and structure have been studied utilizing TEM, SEM, time-lapse micrography during the fixation procedure, volumetry and demonstration of the lysosomal enzyme acid phosphatase. The cells used were in vitro cultivated human glia and glioma cells and suspensions of isolated rat liver parenchymal cells. The fixatives compared were the following: 2 per cent glutaraldehyde (GA) in 0.1 M Na-cacodylate-HCL buffer (cac) with 0.1 M sucrose (pH 7.2); total osmolality (T) 510 mOsmol; vehicle osmolality (V) 300 mOsm, 2 per cent GA in 0.1 M cac (pH 7.2; T = 410 mOsmol; V = 200 mOsmol) and 1.5 per cent GA in 0.067 M cac with 0.033 M sucrose (pH 7.2; T = 320 mOsmol; V = 170 mOsmol). It was found that the fixative with a vehicle osmolality of 300 mOsmol gave results which were interpreted as ideal while the two fixatives were hypotonic vehicles resulted in changes which were easily demonstrated during volumetry, time-lapse micrography, SEM and cytochemistry. However, the differences observed in the TEM were less obvious and difficult to interpret, the major alternations being changes in the configuration of the ER in the liver cells. In conclusion, our findings show that even small variations in the composition of a glutaraldehyde fixative can result in structural changes which do not correspond to the functional morphology of a living cell. Such changes make correct interpretation of micrographs difficult.