Quantitative microbiology with widefield microscopy: navigating optical artefacts for accurate interpretations.

ABSTRACT: Time-resolved live-cell imaging using widefield microscopy is instrumental in quantitative microbiology research. It allows researchers to track and measure the size, shape, and content of individual microbial cells over time. However, the small size of microbial cells poses a significant challenge in interpreting image data, as their dimensions approache that of the microscope's depth of field, and they begin to experience significant diffraction effects. As a result, 2D widefield images of microbial cells contain projected 3D information, blurred by the 3D point spread function. In this study, we employed simulations and targeted experiments to investigate the impact of diffraction and projection on our ability to quantify the size and content of microbial cells from 2D microscopic image

SUBMITTER: Hardo G 

PROVIDER: S-EPMC11368818 | biostudies-literature | 2024

REPOSITORIES: biostudies-literature