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