期刊
JOURNAL OF PHOTOCHEMISTRY AND PHOTOBIOLOGY B-BIOLOGY
卷 99, 期 3, 页码 126-132出版社
ELSEVIER SCIENCE SA
DOI: 10.1016/j.jphotobiol.2010.03.007
关键词
Chromoblastomycosis; Benzazole; Cell staining; Proton transfer; Epifluorescence microscopy; Black fungus
资金
- CNPq
- FAPERGS
The microscopic morphology of Fonsecaea pedrosoi ATCC46428 was observed using two benzazole derivatives, 2-(2'-hydroxyphenyl)benzoxazole and 2-(5'-amino-2'-hydroxyphenyl)benzoxazole, which emit intense fluorescence by a proton transfer mechanism in the electronically excited state (ESIPT). The cell surface could be successfully stained with fluorescent dye solutions of 10 mu M-10 mM using two different fast and cost-effective procedures. At these concentrations, any structure or dye crystallization could be observed. Concerning the external microstructural details, only the amino derivative allowed the differentiation between hyphae and conidia. These dyes presented some advantages comparing to commercial dyes, since the stained cells showed high chemical, thermal and photochemical stability during the experiments and also after several months of storage at room temperature and normal light exposition. Procedure 1 presented the advantage to be used when heating can change the chemical or biochemical cell composition. On the other hand Procedure 2 showed to be useful as a routine methodology for cells staining. The results allowed to propose a simple and highly sensitive assay to study the F. pedrosoi micromorphology by epifluorescence microscopy. This methodology can probably be extended for other fungi of clinical interest. (C) 2010 Elsevier B.V. All rights reserved.
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