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Distribution and behavior of lipid droplets in hepatic cells analyzed by variations of citochemical technique and scanning electron microscopy.
Lima-Faria, João Marcos de; Guimarães, Lucas Nunes; Silva, Victória Costa da; Souza, Iara da Costa; Fernandes, Marisa Narciso; Martinez, Diego Stéfani Teodoro; Sabóia-Morais, Simone Maria Teixeira de.
Afiliación
  • Lima-Faria JM; Laboratory of Cellular Behavior, Institute of Biological Sciences, Federal University of Goiás, Brazil.
  • Guimarães LN; Laboratory of Cellular Behavior, Institute of Biological Sciences, Federal University of Goiás, Brazil.
  • Silva VCD; Laboratory of Cellular Behavior, Institute of Biological Sciences, Federal University of Goiás, Brazil.
  • Souza IDC; Dept. Physiological Science-CCBS, Federal University of São Carlos, Brazil.
  • Fernandes MN; Dept. Physiological Science-CCBS, Federal University of São Carlos, Brazil.
  • Martinez DST; Brazilian Nanotechnology National Laboratory (LNNano), Brazilian Center for Research in Energy and Materials (CNPEM), Campinas, São Paulo, Brazil.
  • Sabóia-Morais SMT; Laboratory of Cellular Behavior, Institute of Biological Sciences, Federal University of Goiás, Brazil.
MethodsX ; 9: 101769, 2022.
Article en En | MEDLINE | ID: mdl-35818446
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.•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.•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.•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.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Tipo de estudio: Prognostic_studies Idioma: En Revista: MethodsX Año: 2022 Tipo del documento: Article País de afiliación: Brasil Pais de publicación: Países Bajos

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Tipo de estudio: Prognostic_studies Idioma: En Revista: MethodsX Año: 2022 Tipo del documento: Article País de afiliación: Brasil Pais de publicación: Países Bajos