Distribution and behavior of lipid droplets in hepatic cells analyzed by variations of citochemical technique and scanning electron microscopy

Author(s)

De Lima-Faria, Joao Marcos ; Guimaraes, Lucas Nunes ; da Silva, Victoria Costa ; Souza, Iara da Costa ; Fernandes, Marisa Narciso ; Teodoro Martinez, Diego Stefani ; Teixeira De Saboia-Morais, Simone Maria

• Document type: Journal article
• Source: METHODSX; v. 9, p. 9-pg., 2022-07-04.

Abstract

Toxicity evaluations involve the analysis of multiple biomarkers. In this study, the liver, target organ analyzed by treatments with iron concentrations, indicated the accumulation of lipids as a response. Considering that the distribution of lipids in an organ is directly related to the induction of inflammatory processes by aquatic contaminants, this study proposes to carry out an integrative investigation of the behavior and the distribution of lipids in the liver tissue. Techniques of light and electron microscopy were performed in order to propose a new way of assessing and quantifying the distribution of lipid droplets, also presenting methodological alternatives that can be chosen by the reader according to the interests and resources available. Thus, it is assumed that the method begins with the fixation of the liver with Glutaraldehyde 2,5% in PBS 0,1 M and continues with post fixation with osmium tretoxide 1%, which marks lipids. For this proposition, two inclusion methodologies were performed to histological analyses in Historesin and ultrastructural analyses in EMBeed 812. For light microscopy (LM) analyses, cuts were obtained with 2,5 micrometers thickness, which were stained with (1) Mayers hematoxylin and (2) toluidine blue. The images obtained were processed in software Image J Fiji to evidence the lipid distribution in liver. center dot Cytological reactions with osmium tetroxide constitute low complexity methods that allow the optimization of the localization, identification and quantification of lipid droplets in the liver tissue when analyzed under the conventional light microscope. center dot Samples included in EMBeed 812 resin commonly used in Transmission Electron Microscopy can be analyzed by SEM-BEC, as complementary analyses for the detection of lipids. center dot Using SEM-BEC and conventional light microscopy, it is possible to quantify the area occupied by lipid droplets using Image J Fiji software, as these are contrasted due to the reaction with osmium tetroxide. (c) 2022 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/) (AU)

• FAPESP's process: 19/24180-4 - Multiple cellular and tissue biomarkers in guppy (Poecilia reticulata) after long-term exposure to glyphosate, LR microscystin and cylinderspermopsin associated to metal nanoparticles (NANOMBI)
• Grantee: Marisa Narciso Fernandes
• Support Opportunities: Regular Research Grants