Lightsheet microscopy, also known as selective plane illumination microscopy (SPIM), is a fluorescence microscopy technique that combines optical sectioning with imaging from multiple views. This facilitates the observation of biological samples and organisms under the principle of tomography for both translucent and opaque samples at different depths. The difference with widefield fluorescence microscopy or confocal microscopy is that lightsheet microscopy only illuminates one optical section at a time, minimizing the effects of photobleaching and phototoxicity. Additionally, the use of computational algorithms allows for three-dimensional reconstruction, even at different time points, generating either a static image or a 3D video as required. The application of this microscopy technique is broad because it can visualize a wide variety of specimens, such as embryos, organoids, spheroids, small animals, small organs, insects, plants, biopsies, etc., which can be cleared or expanded to increase either the depth of view or the resolution.

This is a theoretical-demonstrative course with two main objectives. The first is to introduce lightsheet microscopy; sample preparation through clearing or expansion; and image data management and processing. The second objective is to foster the formation of a regional community of lightsheet microscopy imaging scientists focused on solving scientific and medical problems through the study of the structure and dynamics of three-dimensional biological models that cannot be seen by other microscopy techniques.

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