Distribution and behavior of lipid droplets in hepatic cells analyzed by variations of citochemical technique and scanning electron microscopy.
Distribution and behavior of lipid droplets in hepatic cells analyzed by variations of citochemical technique and scanning electron microscopy.
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DOI：
10.1016/j.mex.2022.101769
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发表时间：
2022
期刊：
影响因子：
1.9
通讯作者：
Teixeira De Saboia-Morais, Simone Maria
中科院分区：
文献类型：
--
作者：
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
关键词：
与四氧化锇的细胞学反应构成了低复杂性的方法，其允许在常规光学显微镜下分析时优化肝组织中脂滴的定位、鉴定和定量。透射电子显微镜中常用的EMBeed 812树脂中包含的样品可以通过SEM-BEC进行分析，作为脂质检测的补充分析。使用SEM-BEC和常规光学显微镜，可以使用Image J Fiji软件量化脂滴所占据的面积，因为这些是由于与四氧化锇的反应而形成对比的。毒性评价涉及多种生物标志物的分析。在这项研究中，肝脏，通过铁浓度处理分析的靶器官，表明脂质的积累是一种反应。考虑到器官中脂质的分布与水生污染物诱导炎症过程直接相关，本研究建议对肝脏组织中脂质的行为和分布进行综合研究。光学和电子显微镜技术进行，以提出一种新的方法来评估和量化的脂滴的分布，也提出了方法的替代品，可以选择由读者根据现有的利益和资源。因此，假设该方法从用PBS 0.1 M中的2.5%戊二醛固定肝脏开始，并继续用标记脂质的1%四氧化锇进行后固定。为此，采用两种纳入方法，在Historesin中进行组织学分析，在EMBeed 812中进行超微结构分析。对于光学显微镜（LM）分析，获得2.5微米厚的切口，其用（1）Mayer苏木精和（2）甲苯胺蓝染色。在Image J Fiji软件中处理获得的图像，以证明肝脏中的脂质分布。
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. 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.
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