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

Publication ,  Journal Article
Fadero, TC; Gerbich, TM; Rana, K; Suzuki, A; DiSalvo, M; Schaefer, KN; Heppert, JK; Boothby, TC; Goldstein, B; Peifer, M; Allbritton, NL ...
Published in: J Cell Biol
May 7, 2018

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.

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Published In

J Cell Biol

DOI

EISSN

1540-8140

Publication Date

May 7, 2018

Volume

217

Issue

5

Start / End Page

1869 / 1882

Location

United States

Related Subject Headings

  • Time-Lapse Imaging
  • Reproducibility of Results
  • Photobleaching
  • Models, Biological
  • Microscopy, Fluorescence
  • Light
  • Imaging, Three-Dimensional
  • Humans
  • Fungi
  • Fluorescence
 

Citation

APA
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Fadero, T. C., Gerbich, T. M., Rana, K., Suzuki, A., DiSalvo, M., Schaefer, K. N., … Maddox, P. S. (2018). LITE microscopy: Tilted light-sheet excitation of model organisms offers high resolution and low photobleaching. J Cell Biol, 217(5), 1869–1882. https://doi.org/10.1083/jcb.201710087
Fadero, Tanner C., Therese M. Gerbich, Kishan Rana, Aussie Suzuki, Matthew DiSalvo, Kristina N. Schaefer, Jennifer K. Heppert, et al. “LITE microscopy: Tilted light-sheet excitation of model organisms offers high resolution and low photobleaching.J Cell Biol 217, no. 5 (May 7, 2018): 1869–82. https://doi.org/10.1083/jcb.201710087.
Fadero TC, Gerbich TM, Rana K, Suzuki A, DiSalvo M, Schaefer KN, et al. LITE microscopy: Tilted light-sheet excitation of model organisms offers high resolution and low photobleaching. J Cell Biol. 2018 May 7;217(5):1869–82.
Fadero, Tanner C., et al. “LITE microscopy: Tilted light-sheet excitation of model organisms offers high resolution and low photobleaching.J Cell Biol, vol. 217, no. 5, May 2018, pp. 1869–82. Pubmed, doi:10.1083/jcb.201710087.
Fadero TC, Gerbich TM, Rana K, Suzuki A, DiSalvo M, Schaefer KN, Heppert JK, Boothby TC, Goldstein B, Peifer M, Allbritton NL, Gladfelter AS, Maddox AS, Maddox PS. LITE microscopy: Tilted light-sheet excitation of model organisms offers high resolution and low photobleaching. J Cell Biol. 2018 May 7;217(5):1869–1882.

Published In

J Cell Biol

DOI

EISSN

1540-8140

Publication Date

May 7, 2018

Volume

217

Issue

5

Start / End Page

1869 / 1882

Location

United States

Related Subject Headings

  • Time-Lapse Imaging
  • Reproducibility of Results
  • Photobleaching
  • Models, Biological
  • Microscopy, Fluorescence
  • Light
  • Imaging, Three-Dimensional
  • Humans
  • Fungi
  • Fluorescence