3D biological imaging using laser-scanning confocal fluorescence microscopy

Abstract

In this work, the three-dimensional (3D) imaging capability of an inexpensive custom-built laser-scanning confocal fluorescence microscope (LSCFM) is demonstrated. Axial resolution is quantified by monitoring the response of the LSCFM to a fluorescence sea and an ultrathin fluorescence monolayer. Correspondingly, we present images of an autofluorescent pollen grain taken at different depths. The 3D volume-rendered image of the specimen is generated from the series of optical sections obtained. Results indicate unique morphology of the optically sectioned biological sample which are not evident in the individual optical sections.