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Light Sheet – Is it all about the Beams

Whether it’s with a Bessel beam, a Gaussian beam, an Airy beam or an Optical Lattice, Light Sheet Fluorescence Microscopy (LSFM) is now an established technique for imaging three dimensional biological samples e.g. tissues, organoids and model organisms like Zebrafish. LSFM provides an effective way to optically section the sample and minimise damage due to illumination, reject out of focus fluorescence and, image quickly and over extended periods of time. Although sample preparation takes extra consideration compared to other approaches, the technique is well suited to live cell imaging, proving very useful in developmental studies of drosophila, c.elegans or zebrafish. The different beam types provide different benefits making them suited to different sample types and there are many self-build and complete solutions available.

Light-sheet fluorescence microscopy imaging of Zebrafish embryo gastruliation from 4 to 18 hours post-fertilization, where each cell nucleus is labelled with GFP. Cells are color-coded for depth to visualisze how dynamic cell reorganization gives rise to the body axis of zebrafish. Image courtesy of Gopi Shah, Max Planck Institute of Molecular Cell Biology and Genetics, Dresden.

sCMOS cameras are a vital element of many LSFM set-ups as they have the required sensitivity, the imaging speed, and large sensor area that facilitates scanning large areas of the specimen at high speeds with minimal bleaching and high signal to noise.

Further Reading

References

  • Tomer R, Lovett-Barron M, Kauvar I, Andalman A, Burns VM, Sankaran S, Grosenick L, Broxton M, Yang S, Deisseroth K. SPED light sheet microscopy: Fast mapping of biological system structure and function. Cell. 2015;163:1796–1806. doi: 10.1016/j.cell.2015.11.061
  • Liebmann T, Renier N, Bettayeb K, Greengard P, Tessier-Lavigne M, Flajolet M. Three-Dimensional Study of Alzheimer's Disease Hallmarks Using the iDISCO Clearing Method. Cell Rep. 2016 Jul 26;16(4):1138-52

Date: February 2020

Author: Dr Alan Mullan and Dr Claudia Florindo

Category: Application Note

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