Light-sheet microscopy or Ultramicroscopy, stands out for its efficient and gentle 3D imaging of large samples, such as an entire mouse, compared to conventional epifluorescent microscopy techniques like widefield or confocal. In light-sheet microscopy, a transparent specimen is illuminated from the side with a thin light sheet. This technique allows for quick optical sectioning and minimal out-of-focus illumination. Photobleaching is reduced, and imaging speed is increased due to the exclusion of out-of-focus light through a pinhole. Modified from Dodt et al. 2007. Dodt et al. 2007 (a) The sample is illuminated from two sides by a blue laser forming a thin sheet of light. Fluorescent light is thus emitted only from a thin optical section and collected by the objective lens. Stray light is blocked by a GFP filter and the image is projected through the tube lens onto the camera target. (b) Photograph of a whole mouse brain in clearing solution illuminated in an early experiment by the white light of a high-pressure mercury burner. Selected literature Dodt, H.-U. et al. Ultramicroscopy: three-dimensional visualization of neuronal networks in the whole mouse brain. Nat. Methods 4, 331–336 (2007).Siedentopf, H. & Zsigmondy, R. U ̈ ber Sichtbarmachung und Gro ̈enbestimmung ultramikroskopischer Teilchen, mit besonderer Anwendung auf Goldrubingla ̈ser. Annalen der Physik 10, 1–39 (1903).Keller, P.J. & Dodt, H.U. Light sheet microscopy of living or cleared specimens. Curr. Opin. Neurobiol. 22, 138–143 (2012).Ueda, H. R. et al. Whole-Brain Profiling of Cells and Circuits in Mammals by Tissue Clearing and Light-Sheet Microscopy. Neuron 106, 369–387 (2020).Becker, K., Jährling, N., Kramer, E.R., Schnorrer, F. & Dodt, H.U. Ultramicroscopy: 3D reconstruction of large microscopical specimens. J. Biophotonics 1, 36–42 (2008). This article was published on 2024-09-09
