title: 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.
authors.affiliation: Department of Microbial and Molecular Pathogenesis, Texas A&M Health Sciences Center, College Station, Texas, USA.
authors.forename: Ying
authors.initials: Y
authors.lastname: Kong
authors.forename: Yanlin
authors.initials: Y
authors.lastname: Shi
authors.forename: Mihee
authors.initials: M
authors.lastname: Chang
authors.forename: Ali R
authors.initials: AR
authors.lastname: Akin
authors.forename: Kevin P
authors.initials: KP
authors.lastname: Francis
authors.forename: Ning
authors.initials: N
authors.lastname: Zhang
authors.forename: Tamara L
authors.initials: TL
authors.lastname: Troy
authors.forename: Hequn
authors.initials: H
authors.lastname: Yao
authors.forename: Jianghong
authors.initials: J
authors.lastname: Rao
authors.forename: Suat L G
authors.initials: SL
authors.lastname: Cirillo
authors.forename: Jeffrey D
authors.initials: JD
authors.lastname: Cirillo
mesh_terms: D000818:Animals; D001419:Bacteria / Q000254:growth & development / Q000378:metabolism / Q000472:pathogenicity*; D001424:Bacterial Infections / Q000473:pathology*; D004195:Disease Models, Animal; D015870:Gene Expression; D017930:Genes, Reporter; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D049449:Luminescence; D049408:Luminescent Agents / Q000378:metabolism; D008163:Luminescent Measurements; D051379:Mice; D051598:Whole Body Imaging / Q000379:methods*; D001618:beta-Lactamases / Q000235:genetics / Q000378:metabolism
chemical_list: D049408:Luminescent Agents; D008156:Luciferases; D001618:beta-Lactamases