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.
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.
2. Aerosol infection of mice using a Madison Chamber
3. Animal anesthesia
4. Reporter enzyme fluorescence (REF) imaging
The authors have nothing to disclose.
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 |