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Using Mycobacterium smegmatis as a Bioindicator for Zinc-Limited Growth Conditions in Mycobacteria08:24

Using Mycobacterium smegmatis as a Bioindicator for Zinc-Limited Growth Conditions in Mycobacteria

This protocol details preparation of Zn2+-limited media, growth vessels, and seed cultures to produce Zn2+-limited ribosomes in mycobacteria. Traces of Zn2+ lead to Zn2+-limiting growth and we describe morphogenic features used as bioindicators to verify Zn2+ limitation in vitro in the model bacterium, Mycobacterium smegmatis, and the human pathogen M.
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Measuring Mycobacterium smegmatis Mistranslation Rates Using a GFP/Luciferase Reporter System03:38

Measuring Mycobacterium smegmatis Mistranslation Rates Using a GFP/Luciferase Reporter System

Source: Chen, Y., et al., Measurement of Specific Mycobacterial Mistranslation Rates with Gain-of-function Reporter Systems. J. Vis. Exp. (2019)This video demonstrates a method to quantify mistranslation in Mycobacterium smegmatis using an inducible GFP and secreted mutant luciferase reporter system. It shows how luminescence resulting from mistranslation is measured and compared to GFP fluorescence to evaluate the impact of a test molecule on translational...
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Customizing a Cryolite Glass Prosthetic Eye08:04

Customizing a Cryolite Glass Prosthetic Eye

This manuscript shows each step of customizing a cryolite glass prosthetic eye including some major advantages of the use of cryolite glass for manufacturing an eye prosthesis compared to poly(methyl methacrylate). In addition, this manuscript gives ophthalmologists better insight into the ocularistic care that could improve interprofessional...
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Flow Cytometry Staining of Mycobacterium tuberculosis Infected Macrophage Subpopulations03:18

Flow Cytometry Staining of Mycobacterium tuberculosis Infected Macrophage Subpopulations

This video showcases a flow cytometric gating strategy utilizing a multi-color panel to analyze cell viability and specific fluorescent-conjugated anti-human antibody markers. This approach makes it possible to determine the percentage of Mycobacterium tuberculosis-infected cell populations...
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An Assay to Evaluate Drug Efficacy against a Mycobacterium tuberculosis Infection Model05:14

An Assay to Evaluate Drug Efficacy against a Mycobacterium tuberculosis Infection Model

This video demonstrates an assay to evaluate the efficacy of drugs against Mycobacterium tuberculosis (Mtb) infection. An in vitro infection model containing Mtb/macrophage aggregates is generated that mimics in vivo infection. Upon introducing suitable antibiotics, their efficacy is tested via resuzarin assay to monitor the survival of bacteria with increasing drug...
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Use of the Invertebrate Galleria mellonella as an Infection Model to Study the Mycobacterium tuberculosis Complex09:23

Use of the Invertebrate Galleria mellonella as an Infection Model to Study the Mycobacterium tuberculosis Complex

Galleria mellonella was recently established as a reproducible, cheap, and ethically acceptable infection model for the Mycobacterium tuberculosis complex. Here we describe and demonstrate the steps taken to establish successful infection of G. mellonella with bioluminescent Mycobacterium bovis BCG...
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