title: Comparison of human optimized bacterial luciferase, firefly luciferase, and green fluorescent protein for continuous imaging of cell culture and animal models.
abstract: Bioluminescent and fluorescent reporter systems have enabled the rapid and continued growth of the optical imaging field over the last two decades. Of particular interest has been noninvasive signal detection from mammalian tissues under both cell culture and whole animal settings. Here we report on the advantages and limitations of imaging using a recently introduced bacterial luciferase (lux) reporter system engineered for increased bioluminescent expression in the mammalian cellular environment. Comparison with the bioluminescent firefly luciferase (Luc) system and green fluorescent protein system under cell culture conditions demonstrated a reduced average radiance, but maintained a more constant level of bioluminescent output without the need for substrate addition or exogenous excitation to elicit the production of signal. Comparison with the Luc system following subcutaneous and intraperitoneal injection into nude mice hosts demonstrated the ability to obtain similar detection patterns with in vitro experiments at cell population sizes above 2.5 × 10(4) cells but at the cost of increasing overall image integration time.
authors.affiliation: University of Tennessee, Center for Environmental Biotechnology, 1414 Circle Drive, 676 Dabney Hall, Knoxville, Tennesssee 37996, USA.
authors.forename: Dan M
authors.initials: DM
authors.lastname: Close
authors.forename: Ruth E
authors.initials: RE
authors.lastname: Hahn
authors.forename: Stacey S
authors.initials: SS
authors.lastname: Patterson
authors.forename: Seung J
authors.initials: SJ
authors.lastname: Baek
authors.forename: Steven A
authors.initials: SA
authors.lastname: Ripp
authors.forename: Gary S
authors.initials: GS
authors.lastname: Sayler
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D003952:Diagnostic Imaging / Q000379:methods*; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D049453:Luciferases, Bacterial / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D049409:Luciferases, Firefly / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D008163:Luminescent Measurements / Q000379:methods*; D008164:Luminescent Proteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008819:Mice, Nude; D012815:Signal Processing, Computer-Assisted; D051598:Whole Body Imaging
chemical_list: D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D049409:Luciferases, Firefly; D049453:Luciferases, Bacterial