BBQ methods: Streamlined workflows for Bacterial Burden Quantification in infected cells by confocal microscopy.

ABSTRACT: Accurate quantification of bacterial burden within macrophages, termed Bacterial Burden Quantification (BBQ), is crucial for understanding host-pathogen interactions. Various methods have been employed, each with strengths and weaknesses. This article addresses limitations in existing techniques and introduces two novel automated methods for BBQ within macrophages based on confocal microscopy data analysis. The first method refines total fluorescence quantification by incorporating filtering steps to exclude uninfected cells, while the second method calculates total bacterial volume per cell to mitigate potential biases in fluorescence-based readouts. These workflows utilize PyImageJ and Cellpose software, providing reliable, unbiased, and rapid quantification of bacterial load. The propos

SUBMITTER: Augenstreich J 

PROVIDER: S-EPMC10592879 | biostudies-literature | 2023 Oct

REPOSITORIES: biostudies-literature