Běhalová Hana, Tremlová Bohuslava, Kalčáková Ludmila, Pospiech Matej, Dordevic Dani
Department of Plant Origin Foodstuffs Hygiene and Technology, Faculty of Veterinary Hygiene and Ecology, University of Veterinary and Pharmaceutical Sciences Brno, 612 42 Brno, Czech Republic.
Department of Technology and Organization of Public Catering, South Ural State University, 454080 Chelyabinsk, Russia.
Foods. 2020 Apr 13;9(4):487. doi: 10.3390/foods9040487.
The aim of the research was to verify the necessity of secondary fixation with osmium tetroxide in various types of meat products and evaluation of structural changes of products using different fixation procedures. The material for the study consisted of 11 types of meat products that were analyzed using a scanning electron microscope (SEM) with two different methods of chemical fixation. The first method included the usual processing of biological samples: glutaraldehyde primary fixation, the use of a buffer, secondary fixation by osmium tetroxide (OsO), buffer, and dehydration using ethanol of increasing concentrations. The second method comprised the glutaraldehyde primary fixation and dehydration using the ethanol of increasing concentrations only. The results unambiguously suggest that the main difference between these methods is in fixation and visibility of fat. Our analysis principally suggests that fixation of the product with OsO allows the tracking of all components (fat droplets, muscle fibers, connective tissue) in meat products. At the same time, our results also support the possibility that the secondary fixation can be skipped during the analysis, where the main objection is an observation of lipid-free structures of the meat products (e.g., connection between muscle and starches or spices) or meat products with an insignificant amount of fat.
该研究的目的是验证在各类肉制品中使用四氧化锇进行二次固定的必要性，并评估使用不同固定程序时产品的结构变化。研究材料包括11种肉制品，采用扫描电子显微镜（SEM）并运用两种不同的化学固定方法对其进行分析。第一种方法包括生物样品的常规处理：戊二醛初次固定、使用缓冲液、四氧化锇（OsO）二次固定、缓冲液处理以及使用浓度递增的乙醇进行脱水。第二种方法仅包括戊二醛初次固定和使用浓度递增的乙醇进行脱水。结果明确表明，这些方法之间的主要差异在于脂肪的固定和可见性。我们的分析主要表明，用OsO固定产品能够追踪肉制品中的所有成分（脂肪滴、肌纤维、结缔组织）。同时，我们的结果也支持在分析过程中可以跳过二次固定的可能性，主要原因是观察无脂的肉制品结构（如肌肉与淀粉或香料之间的连接）或脂肪含量极少的肉制品。
