ESTUDIO ULTRAESTRUCTURAL DE Leptospira biflexa serovar Andamana cepa JNS AL MICROSCOPIO ELECTRÓNICO DE TRANSMISIÓN Y BARRIDO.

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Title: ESTUDIO ULTRAESTRUCTURAL DE Leptospira biflexa serovar Andamana cepa JNS AL MICROSCOPIO ELECTRÓNICO DE TRANSMISIÓN Y BARRIDO.
Authors: Macedo A., Sonia1,2 sma1611@yahoo.com
Source: Revista Peruana de Medicina Experimental y Salud Pública. 2005, Vol. 22 Issue 4, p281-289. 9p.
Subjects: LEPTOSPIRA, TRANSMISSION electron microscopes, SCANNING electron microscopes, CYTOPLASMIC filaments, MORPHOLOGY, MICROORGANISMS
Abstract (English): Objectives: To investigate Leptospira biflexa serovar Andamana, JNS strain ultrastructure using the metallic replication technique, ultrafine cuts, and negative staining for transmission electron microscope and scanning electron microscope high and ultra-high resolution imaging. Materials and methods: Andamana serovar, JNS strain logarithmic phase cultures, 12-15 x 106 cell/mL isolated from one patient were used. Negative staining, ultrafine slicing and metallic replication methods were used for transmission electron microscopy. For scanning electron microscopy, high and ultra-high resolution samples with gold and platinum coating were used. Results: Using the metallic replication method the external structure of individual cells was defined for Leptospira biflexa, Andamana serovar using transmission electron microscopy. Using the negative staining technique the three-layer external membrane could be seen, as well as axial filaments, cytoplasmic membrane, granular cytoplasm with electron-dense inclusions, «balloon» formations, «terminal structure» «terminal appendages», and also the electron-permeable inclusions, all of these were clearly defined. The typical morphology of the microorganism was observed using fine slices on scanning electron microscopy. Conclusions: For the first time we report the metallic replication technique for determining the external morphology in individual Leptospira biflexa, Andamana serovar, JNS strain, and this technique offers advantageous results compared to other methods. Ultrastructure and electron-permeable inclusions within the cytoplasm at a 120 000 power 20 KV were clearly seen using negative staining. Scanning electron microscopy was also useful for confirming the helical structure of the microorganism. [ABSTRACT FROM AUTHOR]
Abstract (Spanish): Objetivos: Investigar la ultraestructura de Leptospira biflexa serovar Andamana cepa JNS, por la técnica de réplica metálica, cortes ultrafinos y tinción negativa al microscopio electrónico de transmisión (MET) y barrido (MEB) en alta y ultraalta resolución. Materiales y métodos: Cultivos en fase logarítmica 12-15x106 cel/mL del serovar Andamana cepa JNS, aislada de un paciente. Se aplicaron los métodos de tinción negativa, cortes ultrafinos y réplica metálica para la microscopía electrónica de transmisión. Para la microscopía electrónica de barrido se usaron las técnicas en alta y ultraalta resolución de muestras cubiertas con oro y platino. Resultados: Con el método de réplica metálica se definió la estructura externa en células individuales de Leptospira biflexa serovar Andamana al MET. Mediante la técnica de tinción negativa se observó la membrana externa formada por tres capas, los filamentos axiales, la membrana citoplasmática, el citoplasma granular con inclusiones electronodensas, las formaciones «globosas», la «estructura terminal», «apéndices terminales», además de las inclusiones permeables para los electrones, claramente definidas. La morfología clásica del microorganismo fue observada en cortes finos, y al microscopio electrónico de barrido. Conclusiones: Se reporta por primera vez la técnica de réplica metálica para determinar la morfología externa en células individuales de Leptospira biflexa serovar Andamana JNS, la cual ofrece resultados ventajosos en comparación con otros métodos. La ultraestructura, y las inclusiones permeables para electrones dentro del citoplasma a una magnificación de 120000x 20KV fueron nítidamente observadas por tinción negativa. La estructura helicoidal del microorganismo se comprobó al MEB. [ABSTRACT FROM AUTHOR]
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Database: MedicLatina
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Abstract:Objectives: To investigate Leptospira biflexa serovar Andamana, JNS strain ultrastructure using the metallic replication technique, ultrafine cuts, and negative staining for transmission electron microscope and scanning electron microscope high and ultra-high resolution imaging. Materials and methods: Andamana serovar, JNS strain logarithmic phase cultures, 12-15 x 106 cell/mL isolated from one patient were used. Negative staining, ultrafine slicing and metallic replication methods were used for transmission electron microscopy. For scanning electron microscopy, high and ultra-high resolution samples with gold and platinum coating were used. Results: Using the metallic replication method the external structure of individual cells was defined for Leptospira biflexa, Andamana serovar using transmission electron microscopy. Using the negative staining technique the three-layer external membrane could be seen, as well as axial filaments, cytoplasmic membrane, granular cytoplasm with electron-dense inclusions, «balloon» formations, «terminal structure» «terminal appendages», and also the electron-permeable inclusions, all of these were clearly defined. The typical morphology of the microorganism was observed using fine slices on scanning electron microscopy. Conclusions: For the first time we report the metallic replication technique for determining the external morphology in individual Leptospira biflexa, Andamana serovar, JNS strain, and this technique offers advantageous results compared to other methods. Ultrastructure and electron-permeable inclusions within the cytoplasm at a 120 000 power 20 KV were clearly seen using negative staining. Scanning electron microscopy was also useful for confirming the helical structure of the microorganism. [ABSTRACT FROM AUTHOR]
ISSN:17264634