TY - JOUR
T1 - Bioluminescence measurements in mice using a skin window
AU - Huang, Qin
AU - Acha, Victor
AU - Yow, Raylon
AU - Schneider, Erik
AU - Sardar, Dhiraj K.
AU - Hornsby, Peter J.
N1 - Funding Information:
This work was supported by a grant from the San Antonio Life Sciences Institute to Hornsby and Sardar. Sardar also acknowledges support from the NSF-sponsored Center for Biophotonics Science and Technology (CBST) at the University of California, Davis, under Cooperative Agreement PHY 0120999.
PY - 2007/9
Y1 - 2007/9
N2 - Studies of bioluminescence in living animals, such as cellbased biosensor applications, require measurement of light at different wavelengths, but accurate light measurement is impeded by absorption by tissues at wavelengths 600 nm. We present a novel approach to this problem-the use of a plastic window in the skin/body wall of mice-that permits measurements of light produced by bioluminescent cells transplanted into the kidney. The cells coexpressed firefly luciferase FLuc, a vasopressin receptor-Renilla luciferase RLuc fusion protein, and a GFP2-arrestin2 fusion protein. Following coadministration of two luciferase substrates, native coelenterazine and luciferin, bioluminescence is measured via the window using fiber optics and a photon counter. Light emission from the two different luciferases, FLuc and RLuc, is readily distinguishable using appropriate optical filters. When coelenterazine 400a is administered, bioluminescence resonance energy transfer BRET occurs between the RLuc and GFP2 fusion proteins and is detected by the use of suitable filters. Following intraperitoneal injection of vasopressin, there is a marked increase in BRET. When rapid and accurate measurement of light from internal organs is required, rather than spatial imaging of bioluminescence, the combination of skin/body wall window and fiber optic light measurement will be advantageous.
AB - Studies of bioluminescence in living animals, such as cellbased biosensor applications, require measurement of light at different wavelengths, but accurate light measurement is impeded by absorption by tissues at wavelengths 600 nm. We present a novel approach to this problem-the use of a plastic window in the skin/body wall of mice-that permits measurements of light produced by bioluminescent cells transplanted into the kidney. The cells coexpressed firefly luciferase FLuc, a vasopressin receptor-Renilla luciferase RLuc fusion protein, and a GFP2-arrestin2 fusion protein. Following coadministration of two luciferase substrates, native coelenterazine and luciferin, bioluminescence is measured via the window using fiber optics and a photon counter. Light emission from the two different luciferases, FLuc and RLuc, is readily distinguishable using appropriate optical filters. When coelenterazine 400a is administered, bioluminescence resonance energy transfer BRET occurs between the RLuc and GFP2 fusion proteins and is detected by the use of suitable filters. Following intraperitoneal injection of vasopressin, there is a marked increase in BRET. When rapid and accurate measurement of light from internal organs is required, rather than spatial imaging of bioluminescence, the combination of skin/body wall window and fiber optic light measurement will be advantageous.
KW - Bioluminescence
KW - Luciferases
KW - Resonance energy transfer
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U2 - 10.1117/1.2795567
DO - 10.1117/1.2795567
M3 - Article
C2 - 17994900
AN - SCOPUS:38449104757
SN - 1083-3668
VL - 12
JO - Journal of biomedical optics
JF - Journal of biomedical optics
IS - 5
M1 - 054012-1
ER -