Polarization effects on fluorescence emission of zebrafish neurons using light-sheet microscopy

TL;DR

This study uses light-sheet microscopy to explore how polarization of excitation light affects fluorescence in zebrafish neurons, revealing polarization-dependent intensity variations and improved imaging contrast.

Contribution

It demonstrates the polarization effects on fluorescence emission in zebrafish neurons and introduces a method to enhance imaging contrast by exploiting polarization differences.

Findings

Fluorescence intensity follows a cosine square dependence on polarization.

Orthogonal polarization yields 40% higher fluorescence emission.

Enhanced signals mainly originate from nerve cells, aiding structural observation.

Abstract

Light-sheet fluorescence microscopy (LSFM) makes use of a thin plane of light to optically section and image transparent tissues or organisms {\it{in vivo}}, which has the advantages of fast imaging speed and low phototoxicity. In this paper, we have employed light-sheet microscopy to investigate the polarization effects on fluorescence emission of zebrafish neurons via modifying the electric oscillation orientation of the excitation light. The intensity of the fluorescence emission from the excited zebrafish larvae follows a cosine square function with respect to the polarization state of the excitation light and reveals a 40$\%$ higher fluorescence emission when the polarization orientation is orthogonal to the illumination and detection axes. Through registration and subtraction of fluorescence images under different polarization states, we have demonstrated that most of the enhanced fluorescence signals are from the nerve cells rather than the extracellular substance. This provides us a way to distinguish the cell boundaries and observe the organism structures with improved contrast and resolution.