This chapter describes the use of whole-body bioluminescent imaging (BLI) for the study of bacterial trafficking in live mice, with an emphasis on the use of bacteria in therapy of cancer. Bacteria present an attractive class of vector for cancer therapy, possessing a natural ability to grow preferentially within tumors following systemic administration. Bacteria engineered to express the lux gene cassette permit BLI detection of the bacteria and tumor sites concurrently. The location and levels of bacteria within tumors over time can be readily examined, visualized in two or three dimensions. The method is applicable to a wide range of bacterial species and tumor xenograft types. This article describes the protocol for analysis of bioluminescent bacteria within subcutaneous tumor-bearing mice. This powerful, and inexpensive, real-time imaging strategy represents an ideal method for the study of bacteria in vivo in the context of cancer research. This protocol outlines the procedure for studying lux-tagged Escherichia coli and Bifidobacterium breve in mice, demonstrating the spatial and temporal readout from 2D and 3D BLI achievable with whole-body in vivo luminescence imaging.
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The authors wish to acknowledge Dr. Cormac Gahan and Dr. Susan Joyce for providing lux-tagged E. coli MG1655 and relevant funding support from the European Commission Seventh Framework Programme (PIAP- GA-2013-612219-VIP), Science Foundation Ireland/Enterprise Ireland (12/TIDA/B2437), the Irish Health Research Board (HRA_POR/2010/138), and the Irish Cancer Society (CRF11CRO and PCI12TAN).
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