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Total internal reflection fluorescence microscopy or TIRF is an advanced microscopic technique used to visualize fluorophores in samples close to a solid surface with a higher refractive index, such as a glass coverslip. TIRF only allows fluorophores in proximity to the solid surface to be excited. When light from a medium with a lower refractive index (such as air) hits the glass coverslip at a critical angle, the light undergoes total internal reflection stead of passing through the glass.
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Using Light Sheet Fluorescence Microscopy to Image Zebrafish Eye Development
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Light sheet fluorescence microscopy for neuroscience.

Stella Corsetti1, Frank Gunn-Moore2, Kishan Dholakia1

  • 1Scottish Universities Physics Alliance, School of Physics and Astronomy, University of St. Andrews, North Haugh, St. Andrews, Fife KY16 9SS, UK.

Journal of Neuroscience Methods
|July 27, 2018
PubMed
Summary
This summary is machine-generated.

Light sheet fluorescence microscopy (LSFM) offers high spatiotemporal resolution for imaging neuronal networks in the central nervous system (CNS). This advanced technique enables in-vivo imaging and detailed morphological studies of the brain.

Keywords:
Airy beamsBessel beamsBrainLSFMLight sheet fluorescence microscopyNeuroimagingNeuroscienceSPIM

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Area of Science:

  • Neuroscience
  • Biomedical Imaging
  • Optical Microscopy

Background:

  • Central nervous system (CNS) functions depend on large-scale neuronal network interactions.
  • High spatiotemporal resolution imaging is crucial for understanding CNS functions.

Purpose of the Study:

  • To highlight the need for advanced imaging techniques in neuroscience.
  • To present Light Sheet Fluorescence Microscopy (LSFM) as a solution.

Main Methods:

  • Light Sheet Fluorescence Microscopy (LSFM) for volumetric imaging.
  • Use of propagation invariant Bessel and Airy beams to enhance penetration depth.

Main Results:

  • LSFM provides high spatial resolution and minimizes photobleaching/phototoxicity.
  • Enabled in-vivo imaging of CNS development and sub-cellular resolution of cleared brains.
  • Propagation invariant beams improve imaging depth in neural tissues.

Conclusions:

  • LSFM is a promising technique for in-vivo functional and morphological imaging of the CNS.
  • Further developments in LSFM geometries and beam types could enable deeper in-vivo imaging.