Reporter Enzyme Fluorescence Imaging for In Vivo Tracking of Bacterial Infection

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.

1. Grow the bacteria in M-OADC-TW (7H9 broth supplemented with 0.5% glycerol, 10% oleic acid dextrose complex without catalase, and 0.05% Tween-80) medium standing at 37 °C to an OD₆₀₀ of 0.5 (~ 0.2 x 10⁷ colony-forming unit or CF....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
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