Optimal clearing and mounting media for confocal microscopy of thick specimens
نویسندگان
چکیده
Sample preparation for confocal microscopy typically involves fixing and staining, followed by clearing and mounting. Clearing involves treatment with solvent to dissolve lipids and other structures that cause light scattering and reduce the ability to image through thick specimens. Mounting involves transfer of the specimen into a medium which matches the refractive index the objective is designed for (n = 1.515 in the case of oil immersion lenses). This higher refractive index also helps to match the refractive index of the tissue. Mounting media often include chemicals to retard photobleaching. A wide variety of clearing and mounting media for confocal microscopy have been described in the literature; in several cases the same media are used for both clearing and mounting. Careful attention to clearing and mounting has enabled confocal microscopy of samples greater than 500 μm thick at low magnification [2, 11-12].
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