Bioluminescence imaging

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Bioluminescence imaging (BLI) is a technology developed over the past decade that allows for the noninvasive study of ongoing biological processes in small laboratory animals.

Bioluminescence is the process of light emission in living organisms. Bioluminescence imaging utilizes native light emission from one of several organisms which bioluminesce. The three main sources are the North American firefly, the sea pansy (and related marine organisms), and bacteria like Photorhabdus luminescens and Vibrio fischeri. The DNA encoding the luminescent protein is incorporated into the laboratory animal either via a virus or by creating a transgenic animal.

Systems derived from the three groups above differ in key ways:

1. Firefly luciferase requires D-luciferin to be injected into the subject prior to imaging. The peak emission wavelength is about 560 nm. Due to the attenuation of blue-green light in tissues, the red-shift (compared to the other systems) of this emission makes detection of firefly luciferase much more sensitive in vivo.
2. Renilla luciferase (from the Sea Pansy) requires its substrate, coelenterazine, to be injected as well. As opposed to luciferin, coelenterazine has a lower bioavailability (likely due to MDR1 transporting it out of mammalian cells). Additionally, the peak emission wavelength is about 480 nm.
3. Bacterial luciferase has an advantage in that the lux  operon used to express it also encodes the enzymes required for substrate biosynthesis. Unfortunately, this system has not yet been adapted for mammalian cell expression (but is widely used for developing bioluminescent pathogens). This luciferase reaction has a peak wavelength of about 490 nm.

While the total amount of light emitted from bioluminescence is typically small and not detected by the human eye, an ultra-sensitive CCD camera can image bioluminescence from an external vantage point.

Common applications of BLI include in vivo studies of infection (with bioluminescent pathogens), cancer progression (using a bioluminescent cancer cell line), and reconstitution kinetics (using bioluminescent stem cells).

Researchers at UT Southwestern Medical Center have shown that bioluminescence imaging can be used to determine the effectiveness of cancer drugs that choke off a tumor’s blood supply. The technique requires luciferin to be added to the bloodstream, which carries it to cells throughout the body. When luciferin reaches cells that have been altered to carry the firefly gene, those cells emit light.^[1]

[edit] References

• Hutchens, M. and Luker, G.D. (2007). Applications of bioluminescence imaging to the study of infectious diseases. Cellular Microbiology  9, 2315-2322.