Biology 2382B Lecture Notes - Primary And Secondary Antibodies, Sound, Fluorophore

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3: isolation & analysis of organelles & molecules. Confocal fluorescence microscopy illuminates only a focal plane of the specimen and achieves greater clarity than conventional fluorescence microscopy, which produces a blurry image due to fluorescence detection above and below the focal plane. Deconvolution is a computational restoration method (using the point spread function algorithm) that provides an alternative means of producing high-resolution images. In deconvolution microscopy, a series of images of an object are taken at different focal planes called a z-stack with a conventional fluorescence microscope or confocal microscope. Unlike confocal techniques, which physically remove the out-of-focus emission information by means of a pinhole, deconvolution is a mathematical processing method in which computations for the acquired stacks reassign the diffracted light to its original location. As the emitted signal is collected in its entirety by means of a highly sensitive digital camera, post-deconvolution images may in some cases provide higher resolution than confocal microscopy.

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