Brain Structure Morphology After Being Fixated With Ethanol on Electron Microscope

Abstract

Fixation is one of the processes in preparing histology and pathology. The common material for fixation is buffered formalin including paraformaldehyde. However, the effect of the damaged cells, which is fixed for a long time, causes the research for other fixation materials to become necessary. In addition, paraformaldehyde is also harmful to human body and natural environment. Ethanol is one of the alternative fixation materials, which has been used for two hundred years. It has been used for many purposes, both in routine staining and immunohistochemistry. Nonetheless, no research confirms its effect on the electron microscope. The authors studied the effect of 50 % of ethanol on the cell membrane, organelles, and nucleus of Purkinje cells (Neuron purkinjense) observed on a light microscope and Transmitted Electron Microscope (TEM). Then it was compared to buffered formalin. In the light microscope, it shows that both of fixations have no different effects of the morphology of the cell membrane, cytoplasm, the nucleus of Purkinje cells and the neutrophils. We assume that our 50 % of ethanol concentration is almost the same as BF 10 % in the ability of hardening tissue and color absorption based on the previous study. In TEM, the structure of the cell membrane, organelles, and cytoplasm of Purkinje cell look broken in the cerebellum of 50 % of ethanol except for the nucleus. There was no significant difference diameter of the nucleus. It happened in general because of the shrinkage effect of ethanol. However, the authors recommend using 50 % of ethanol for routine staining.

Introduction of Histopathology

“…Research conducted by Arni et al states that alcohol fixation can be done with ethanol with gradual concentrations, from 40% for 24 hours and up to 60% within the next 24 hours. The results of this study showed a good contour cell seen in the light microscope [11].…”

“…NBF 10% or FFPE is used routinely for specimens viewed under the light microscope. Meanwhile, glutaraldehyde, or a mixture of formaldehyde and glutaraldehyde and osmium tetraoxide is used routinely for specimens viewed under an electron microscope [3,11]. Alcohol can also be used for fixation, but it is necessary to pay attention to its percentage and fixation time.…”