STAR protocols | VOL. 3
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Medium throughput protocol for genome-based quantification of intracellular mycobacterial loads and macrophage survival during invitro infection.

Publication Date Jun 17, 2022

Abstract

Here, we present a streamlined protocol for assessing intracellular Mycobacterium tuberculosis (Mtb) loads in macrophages. This protocol describes the simultaneous assessment of macrophage viability using automated microscopy. Further, we detail the quantification of mycobacterial loads using a rapid, inexpensive, and accurate approach for mycobacterial DNA isolation from paraformaldehyde-fixed macrophages. Simultaneous assessment of the bacterial loads using internal standard and macrophage viability allows for precise quantification of the effects of perturbations on Mtb and host cells while accounting for technical artifacts. For complete details on the use and execution of this protocol, please refer to Chatterjee etal. (2021).

Concepts

Quantification Of Loads Invitro Infection Mycobacterial Loads Macrophage Viability Macrophage Survival Mtb Loads Protocol For Quantification Mtb Cells Internal Standard Intracellular Macrophage

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No potential conflict of interest was reported by the authors. The conception and design of the study, acquisition of data, analysis and interpretatio...

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