LITE microscopy: Tilted light-sheet excitation of model organisms offers high resolution and low photobleaching

The Journal of Cell Biology
Tanner C FaderoPaul S Maddox

Abstract

Fluorescence microscopy is a powerful approach for studying subcellular dynamics at high spatiotemporal resolution; however, conventional fluorescence microscopy techniques are light-intensive and introduce unnecessary photodamage. Light-sheet fluorescence microscopy (LSFM) mitigates these problems by selectively illuminating the focal plane of the detection objective by using orthogonal excitation. Orthogonal excitation requires geometries that physically limit the detection objective numerical aperture (NA), thereby limiting both light-gathering efficiency (brightness) and native spatial resolution. We present a novel live-cell LSFM method, lateral interference tilted excitation (LITE), in which a tilted light sheet illuminates the detection objective focal plane without a sterically limiting illumination scheme. LITE is thus compatible with any detection objective, including oil immersion, without an upper NA limit. LITE combines the low photodamage of LSFM with high resolution, high brightness, and coverslip-based objectives. We demonstrate the utility of LITE for imaging animal, fungal, and plant model organisms over many hours at high spatiotemporal resolution.

References

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Citations

Apr 15, 2018·The Journal of Cell Biology·Simon C Watkins, Claudette M St Croix
Oct 9, 2019·Journal of Biophotonics·Yuhao LiuShiwen Zhang
Oct 28, 2019·The Journal of Cell Biology·Alexandra BondazMonica Gotta
Feb 23, 2020·Platelets·Abdullah O Khan, Jeremy A Pike
Apr 4, 2020·Annual Review of Biomedical Engineering·Sheng LiuFang Huang
Jul 17, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Alexander W A F ReismannGerhard J Schütz
Feb 15, 2019·Frontiers in Neuroanatomy·Ignacio Albert-SmetJorge Ripoll
Nov 6, 2018·Cold Spring Harbor Protocols·Bob Goldstein
May 9, 2019·Biomedical Optics Express·Aleks PonjavicDavid Klenerman
Jan 16, 2019·Journal of the Optical Society of America. A, Optics, Image Science, and Vision·Mouna HaouasIlya Golub
Mar 23, 2019·Frontiers in Genetics·Sarah C Hughes, Andrew J Simmonds
Dec 2, 2020·Optics Letters·Ting-Yu HsiehYuan Luo
Feb 18, 2021·Pain Medicine : the Official Journal of the American Academy of Pain Medicine·Adam M R GrohCheryle A Séguin
May 20, 2021·Proceedings of the National Academy of Sciences of the United States of America·Lyndsay L LeachJeffrey M Gross
Jun 15, 2021·Lab on a Chip·Erick J Vargas-OrdazVictor J Cadarso
Dec 1, 2019·Nanophotonics·Jialei TangKyu Young Han

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Methods Mentioned

BETA
confocal microscopy
fluorescence microscopy
LITE microscopy
dissection
biopsy
dissecting

Software Mentioned

Elements
AutoCAD
TILT3D
LITE
Fiji
NIS
Kymograph

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