Sarmiento-Lacera Ladys, Torres-Fernández Orlando, Rivera Jorge A, Rodríguez-Toro Gerzaín
Grupo de Morfología Celular, Dirección de Investigación en Salud Pública, Instituto Nacional de Salud, Bogotá, Colombia.
Facultad de Medicina, Universidad de la Sabana, Bogotá, Colombia.
Microsc Res Tech. 2021 Apr;84(4):789-795. doi: 10.1002/jemt.23638. Epub 2020 Nov 11.
Lobomycosis is a skin infection produced by the fungus Lacazia loboi, which mainly affects some indigenous and afro-descendant populations in Tropical America. We previously reported the comparative effect of osmium tetroxide (OsO ) and ruthenium tetroxide (RuO ) in the electron microscopy (EM) of other related microorganisms. The objective of this study is to compare the effect of postfixation with OsO and RuO in the ultrastructure of L. loboi yeasts. Skin biopsies on patients diagnosed with lobomycosis were fixed in glutaraldehyde at 3% and postfixed in the following solutions: (a) 1% OsO , (b) 0.2% RuO , and (c) OsO at 1% followed by RuO at 0.2%. They were then processed using the conventional method for EM. Unlike OsO the treatment with RuO revealed different shades of gray and electron dense bands in the cell wall and other cell components of L. loboi. The most notable finding was the presence of radial filamentous structures around the yeast, which made the image look like the sun. Postfixation with RuO revealed ultrastructural details that had not been previously reported for L loboi. The combined use of OsO and RuO in EM of microorganisms with cell walls can be useful to evaluate the effect of microbicide substances.
芽生菌病样肉芽肿是由罗博隐球菌引起的一种皮肤感染,主要影响热带美洲的一些原住民和非洲裔人群。我们之前报道过四氧化锇(OsO₄)和四氧化钌(RuO₄)在其他相关微生物电子显微镜检查中的比较效果。本研究的目的是比较用OsO₄和RuO₄进行后固定对罗博隐球菌酵母超微结构的影响。对诊断为芽生菌病样肉芽肿的患者进行皮肤活检,用3%的戊二醛固定,然后在以下溶液中进行后固定:(a)1%的OsO₄,(b)0.2%的RuO₄,(c)1%的OsO₄后接0.2%的RuO₄。然后采用常规的电子显微镜方法进行处理。与OsO₄不同,用RuO₄处理后,罗博隐球菌的细胞壁和其他细胞成分呈现出不同深浅的灰色和电子致密带。最显著的发现是酵母周围存在放射状丝状结构,使图像看起来像太阳。用RuO₄进行后固定揭示了罗博隐球菌以前未报道过的超微结构细节。在对有细胞壁的微生物进行电子显微镜检查时,联合使用OsO₄和RuO₄有助于评估杀微生物物质的效果。