Monday, June 14, 2010

SUPERMICROSCOPES










The Max Planck Institute in collaboration with the company Leica develops new microscopes, able to solve images to scale of some few nanómeters, with a resolution but there the limits of diffraction (nanoscopes). These microscopes based on principles of Photoactivation Localization Microscopy (PALM) and Stimulated Emission Depletion (STED), developed in 1994 by Stefan Hell (Max Planck Institute/Göttingen/To solve images very near at nanoscale range), focusing the opening light with 2 opposed lenses (4Pi Microscope), improving 7 times the space resolution, providing images of high resolution of the position of thousands of molecules.

Technology improved when being created later the GSDIM (Ground state depletion microscopy followed by individual molecule return), adding to STED rays laser activators of fluorescent tints, resulting biomolecular images of great sharpness at nanometer range. Tints include fluorophores, rhodamines and photoactivated fluorescent proteins. Activated individual molecules return spontaneously to their non fluorescent state, while their neighboring counterparts remain non illuminated. Methods developed lately include FPALM (Fluorescence Photoactivation Localization Microscopy), STORM (Stochastic Optical Reconstruction MIcroscopy) and 3D STORM, with which microtubules, DNA complex, movements of proteins and synaptic vesicles in alive cells, mitochondrias, dentrites, etc.,can be seen.
SUPERMICROSCOPIOS

El Instituto Max Planck en colaboracion con la compañia Leica desarrolla nuevos microscopios, capaces de resolver imágenes a escala de unos pocos nanómetros, con una resolucion mas alla del limite de difraccion (nanoscopios). Estos microscopios se apoyan en principios derivados del Photoactivation Localization Microscopy (PALM) y el Stimulated Emission Depletion (STED), desarrollada en 1994 por Stefan Hell (Max Planck Institute/Göttingen/Para resolver imágenes a nanoescala muy cercanas entre si), focalizando la luz de apertura con 2 lentes opuestos (Microscopio 4Pi), mejorando 7 veces la resolución espacial, proporcionando imagenes de alta resolucion de la posición de miles de moleculas.
Tecnologia mejorada al crearse posteriormente el GSDIM (Ground state depletion microscopy followed by individual molecule return), al añadir al STED rayos laser activadores de tinciones fluorescentes, proporcionado imagenes biomoleculares de gran agudeza en el rango de nanometros. Las tinciones incluyen fluoroforos, rodaminas y proteínas fluorescentes fotoactivables. Las moléculas individuales activadas retornan espontaneamente a su estado inicial no fluorescente, mientras sus contrapartes vecinas permanecen no iluminadas. Metodos desarrollados ultimamente incluyen al FPALM (Fluorescence Photoactivation Localization Microscopy), al STORM (Stochastic Optical Reconstruction MIcroscopy) y al 3D STORM, con los que se pueden ver microtubulos, complejos de DNA, movimientos de proteínas y vesículas sinápticas en células vivas, mitocondrias, dentritas,etc.


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