Principios y aplicaciones de la microscopia láser confocal en la investigación biomédica
Palabras clave:
Microscopia láser confocal, fluorescencia, confocalidad, Z-stack, imagen 3DResumen
La microscopia confocal es una herramienta ampliamente utilizada y de apoyo en el área biomédica que ofrece la posibilidad de obtener imágenes con una alta resolución espacial. Lo anterior se debe al principio de confocalidad que permite dirigir el haz de luz láser que ilumina la muestra a un plano focal limitado por el usuario, lo que evita que la adquisición de información quede fuera de foco, lo que mejora significativamente la resolución espacial en particular en el plano «z» (plano de profundidad) de una imagen, generando información más precisa que la que genera un sistema de fluorescencia convencional. Con la microscopia confocal es posible obtener y analizar imágenes tanto de cultivos celulares, como de especímenes con un grosor en el rango de los cientos de micras tales como secciones de tejido que contengan células y moléculas autofluorescentes o marcadas con fluoróforos. Su principal ventaja radica en la obtención de imágenes nítidas de planos correspondientes a 0.5 μm de grosor y que en conjunto con programas de imágenes de libre acceso, pueden realizarse reconstrucciones y análisis en tres dimensiones de las imágenes adquiridas. El constante desarrollo tecnológico ha facilitado el uso de la técnica y el desarrollo de nuevas aplicaciones en áreas de la biomedicina, la biología y la biotecnología. El propósito de esta revisión es dar a conocer las aplicaciones de la microscopia confocal en el campo de la biomedicina, exponiendo las ventajas que presenta con respecto a la microscopia de fluorescencia convencional y mostrar que constituye
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Derechos de autor 2016 Instituto Nacional de Rehabilitación Luis Guillermo Ibarra Ibarra
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