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|1. (WO2018089948) SYSTEM AND METHOD FOR LIGHT SHEET MICROSCOPE AND CLEARING FOR TRACING|
|Applicants:||COLD SPRING HARBOR LABORATORY
VENKATARAJU, Kannan, Umadevi
ALBEANU, Dinu, F.
|Title:||SYSTEM AND METHOD FOR LIGHT SHEET MICROSCOPE AND CLEARING FOR TRACING|
An exemplary system and method for imaging tissue includes using an illumination objective, directing one or multi photon excitation lights onto a portion of a tissue from a position on top and at an oblique angle relative to the tissue while the tissue is mounted on a stage. The method further includes generating a tissue-penetrating light-sheet from the one or multi photon excitation lights. Using a detection objective, the method detects the tissue-penetrating light-sheet. Upon detecting the tissue-penetrating light-sheet, it uses the detection objective, to collect fluorescent signals from the tissue and uses the fluorescent lights to acquire a first image of the tissue while the tissue is an imaging position. A second image of the tissue is acquired while the tissue is in the imaging position. The first and second images each defined by first and second data, respectively. Subsequently, the tissue is moved to a sectioning position and with the use of an integrated Vibratome, a portion of the tissue, with known thickness, is sectioned. The process repeats until the tissue, in its entirety, has been sectioned, with images acquired each time. Image data, from the acquired images, are stitched to create a 3-dimensional 3D) image of the tissue.