Quantifiable Intravital Light Sheet Microscopy

Methods Mol Biol. 2022:2440:181-196. doi: 10.1007/978-1-0716-2051-9_11.

Abstract

Live imaging of zebrafish embryos that maintains normal development can be difficult to achieve due to a combination of sample mounting, immobilization, and phototoxicity issues that, once overcome, often still results in image quality sufficiently poor that computer-aided analysis or even manual analysis is not possible. Here, we describe our mounting strategy for imaging the zebrafish midbrain-hindbrain boundary (MHB) with light sheet fluorescence microscopy (LSFM) and pilot experiments to create a study-specific set of parameters for semiautomatically tracking cellular movements in the embryonic midbrain primordium during zebrafish segmentation.

Keywords: Bioimage analysis; Light sheet fluorescence microscopy; Midbrain–hindbrain boundary; Zebrafish.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Mesencephalon
  • Microscopy, Fluorescence
  • Rhombencephalon
  • Zebrafish Proteins*
  • Zebrafish*

Substances

  • Zebrafish Proteins