Whole‐Body Imaging of Infection Using Bioluminescence

Abstract

Bioluminescence imaging is a powerful technique to visualize and monitor biological processes in numerous systems. This unit describes two strategies for bioluminescence imaging that can be used to study bacterial infection in mice. One method is to express a luciferase gene in the bacteria; the second method is to use bacteria that express both a luciferase and β‐lactamase along with a substrate containing caged luciferin, which is released by β‐lactamase hydrolysis and reacts with luciferase to generate light. For both strategies, bioluminescent signals are imaged using an IVIS live animal imaging system (Caliper Life Sciences). The bioluminescence images are analyzed to localize bioluminescent bacteria, quantify signal, and determine the wavelengths of the signals produced. The correlation of bacterial numbers with signal intensity in vivo can be determined, allowing a quantitative measure of bacterial numbers in mice in real time. Methods are described in detail to facilitate successful application of these emerging technologies in nearly any experimental system. Curr. Protoc. Microbiol. 21:2C.4.1‐2C.4.17. © 2011 by John Wiley & Sons, Inc.

Bioluminescent imaging of Borrelia burgdorferi in vivo demonstrates that the fibronectin‐binding protein BBK32 is required for optimal infectivity

“…Each sample for each strain was treated with a final concentration of 667 µM d ‐luciferin (Research Products International Corp., Mt. Prospect, IL) in PBS and immediately measured for luminescence (Kong et al ., 2011). Each cell concentration for all the strains was measured for luminescence in triplicate, values were averaged and the standard error was calculated.…”

“…The use of in vivo imaging has been adapted to globally track various target cell types as well as the infectious potential of a number of pathogens that have been modified to display fluorescence or bioluminescence (Contag et al ., 1995; Francis et al ., 2000; 2001; Burns et al ., 2001; Edinger et al ., 2002; Rhee et al ., 2010; Kong et al ., 2011). This technology has the advantage of allowing temporal and spatial evaluation of an infectious process in a non‐invasive manner whereby the same animal in its entirety can been analysed (under anaesthesia) at numerous times throughout an experiment.…”

Bacterial vectors for imaging and cancer gene therapy: a review

“…97 Bluco is a 'caged' luciferin, which is released by bacterial b-lactamase hydrolysis for reacting with luciferase. 98 Tumor targeting …”

In vivoimaging of bioluminescentPseudomonas aeruginosain an acute murine airway infection model

“…Bioluminescence imaging allows the analysis of bacterial infectious diseases in animal models Kong et al, 2011). During the last 20 years, non-invasive imaging studies have been performed with numerous micro-organisms including Pseudomonas aeruginosa (reviewed by Andreu et al, 2011).…”