Reporter Enzyme Fluorescence Imaging for In Vivo Tracking of Bacterial Infection

Published: November 30, 2023

Abstract

Source: Sharan, R., et al. Imaging Mycobacterium tuberculosis in Mice with Reporter Enzyme Fluorescence. J. Vis. Exp. (2018).

This video demonstrates the reporter enzyme fluorescence (REF) imaging for in vivo detection of Mycobacterium tuberculosis infection. The infected mice are intraperitoneally injected with a substrate for REF imaging. Upon being hydrolyzed by bacterial β-lactamase enzyme, the substrate emits fluorescence, the detection of which aids evaluation of infection.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Strains and culture conditions Note: M. tuberculosis strain CDC1551 is used in this study, but any M. tuberculosis strain can be used in the same manner. Grow the bacteria in M-OADC-TW (7H9 broth supplemented with 0.5% glycerol, 10% oleic acid dextrose compl…

Declarações

The authors have nothing to disclose.

Materials

Isoflurane VETONE 501027
CNIR800 Custom synthesized
Fatal Plus solution Vortech Pharmaceutical Ls, Ltd
7H9 Middlebrook broth BD 271310
OADC Middlebrook enrichment BD 212351
Sporcidin RE-1284F
7H11 Middlebrook Agar BD 212203
Madison Chamber
IVIS Spectrum Perkin Elmer 124262
XGI-8-gas Anesthesia System Perkin Elmer
Living Imaging software Perkin Elmer
Transparent nose cones Perkin Elmer
M. tuberculosis strain CDC1551 ATCC
Female BALB/C mice, 5-7 weeks Jackson Laboratory

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Reporter Enzyme Fluorescence Imaging for In Vivo Tracking of Bacterial Infection. J. Vis. Exp. (Pending Publication), e21791, doi: (2023).

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