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Technical implementations of light sheet microscopy

Elisa Zagato (UGent) , Toon Brans (UGent) , Stefaan De Smedt (UGent) , Katrien Remaut (UGent) , Kristiaan Neyts (UGent) and Kevin Braeckmans (UGent)
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Abstract
Fluorescence-based microscopy is among the most successful methods in biological studies. It played a critical role in the visualization of subcellular structures and in the analysis of complex cellular processes, and it is nowadays commonly employed in genetic and drug screenings. Among the fluorescence-based microscopy techniques, light sheet fluorescence microscopy (LSFM) has shown a quite interesting set of benefits. The technique combines the speed of epi-fluorescence acquisition with the optical sectioning capability typical of confocal microscopes. Its unique configuration allows the excitation of only a thin plane of the sample, thus fast, high resolution imaging deep inside tissues is nowadays achievable. The low peak intensity with which the sample is illuminated diminishes phototoxic effects and decreases photobleaching of fluorophores, ensuring data collection for days with minimal adverse consequences on the sample. It is no surprise that LSFM applications have raised in just few years and the technique has been applied to study a wide variety of samples, from whole organism, to tissues, to cell clusters, and single cells. As a consequence, in recent years numerous set-ups have been developed, each one optimized for the type of sample in use and the requirements of the question at hand. Hereby, we aim to review the most advanced LSFM implementations to assist new LSFM users in the choice of the LSFM set-up that suits their needs best. We also focus on new commercial microscopes and do-it-yourself strategies; likewise we review recent designs that allow a swift integration of LSFM on existing microscopes.
Keywords
PLANE ILLUMINATION MICROSCOPY, CROSS-CORRELATION SPECTROSCOPY, SELF-RECONSTRUCTING BEAMS, THICK BIOLOGICAL SAMPLES, CELL 3D SUPERRESOLUTION, FLUORESCENCE MICROSCOPY, HIGH-SPEED, ZEBRAFISH DEVELOPMENT, FLOW-CYTOMETRY, BESSEL BEAMS

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Citation

Please use this url to cite or link to this publication:

Chicago
Zagato, Elisa, Toon Brans, Stefaan De Smedt, Katrien Remaut, Kristiaan Neyts, and Kevin Braeckmans. 2018. “Technical Implementations of Light Sheet Microscopy.” Microscopy Research and Technique 81 (9): 941–958.
APA
Zagato, E., Brans, T., De Smedt, S., Remaut, K., Neyts, K., & Braeckmans, K. (2018). Technical implementations of light sheet microscopy. MICROSCOPY RESEARCH AND TECHNIQUE, 81(9), 941–958.
Vancouver
1.
Zagato E, Brans T, De Smedt S, Remaut K, Neyts K, Braeckmans K. Technical implementations of light sheet microscopy. MICROSCOPY RESEARCH AND TECHNIQUE. WILEY; 2018;81(9):941–58.
MLA
Zagato, Elisa et al. “Technical Implementations of Light Sheet Microscopy.” MICROSCOPY RESEARCH AND TECHNIQUE 81.9 (2018): 941–958. Print.
@article{8608884,
  abstract     = {Fluorescence-based microscopy is among the most successful methods in biological studies. It played a critical role in the visualization of subcellular structures and in the analysis of complex cellular processes, and it is nowadays commonly employed in genetic and drug screenings. Among the fluorescence-based microscopy techniques, light sheet fluorescence microscopy (LSFM) has shown a quite interesting set of benefits. The technique combines the speed of epi-fluorescence acquisition with the optical sectioning capability typical of confocal microscopes. Its unique configuration allows the excitation of only a thin plane of the sample, thus fast, high resolution imaging deep inside tissues is nowadays achievable. The low peak intensity with which the sample is illuminated diminishes phototoxic effects and decreases photobleaching of fluorophores, ensuring data collection for days with minimal adverse consequences on the sample. It is no surprise that LSFM applications have raised in just few years and the technique has been applied to study a wide variety of samples, from whole organism, to tissues, to cell clusters, and single cells. As a consequence, in recent years numerous set-ups have been developed, each one optimized for the type of sample in use and the requirements of the question at hand. Hereby, we aim to review the most advanced LSFM implementations to assist new LSFM users in the choice of the LSFM set-up that suits their needs best. We also focus on new commercial microscopes and do-it-yourself strategies; likewise we review recent designs that allow a swift integration of LSFM on existing microscopes.},
  author       = {Zagato, Elisa and Brans, Toon and De Smedt, Stefaan and Remaut, Katrien and Neyts, Kristiaan and Braeckmans, Kevin},
  issn         = {1059-910X},
  journal      = {MICROSCOPY RESEARCH AND TECHNIQUE},
  keywords     = {PLANE ILLUMINATION MICROSCOPY,CROSS-CORRELATION SPECTROSCOPY,SELF-RECONSTRUCTING BEAMS,THICK BIOLOGICAL SAMPLES,CELL 3D SUPERRESOLUTION,FLUORESCENCE MICROSCOPY,HIGH-SPEED,ZEBRAFISH DEVELOPMENT,FLOW-CYTOMETRY,BESSEL BEAMS},
  language     = {eng},
  number       = {9},
  pages        = {941--958},
  publisher    = {WILEY},
  title        = {Technical implementations of light sheet microscopy},
  url          = {http://dx.doi.org/10.1002/jemt.22981},
  volume       = {81},
  year         = {2018},
}

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