Fast imaging and dynamics of the DAPI dye in the live cells using Bessel beam light sheet microscopy

Abstract

Fluorescence microscopy is one of the most widely used technique in biology for imaging. The biological specimen or the area of interest is labelled with suitable dyes or fluorescent nanoparticles for imaging. Most of the standard fluorescence microscopes use an epi-illumination configuration that illuminates the entire sample volume during imaging, and information is collected from a small sample volume. This overexposure of light to the whole of specimen volume can create photobleaching and phototoxicity to live cells. In confocal microscopy, it employs point illumination and detection, which will slow down the imaging process. Fast imaging is necessary to study the cellular dynamics on a microscopic scale. A Bessel beam-based light sheet microscopy can overcome these limitations. A Bessel beam-based light sheet microscope uses an orthogonal configuration of excitation and detection arms, which will reduce the volume of illumination and reduce photobleaching. Since light sheet microscopy employs line scanning for excitation, time-lapse images of hundreds of frames per second can be obtained using a CCD camera.