title: iRFP is a sensitive marker for cell number and tumor growth in high-throughput systems.
abstract: GFP and luciferase are used extensively as markers both in vitro and in vivo although both have limitations. The utility of GFP fluorescence is restricted by high background signal and poor tissue penetrance. Luciferase throughput is limited in vitro by the requirement for cell lysis, while in vivo, luciferase readout is complicated by the need for substrate injection and the dependence on endogenous ATP. Here we show that near-infrared fluorescent protein in combination with widely available near-infrared scanners overcomes these obstacles and allows for the accurate determination of cell number in vitro and tumor growth in vivo in a high-throughput manner and at negligible per-well costs. This system represents a significant advance in tracking cell proliferation in tissue culture as well as in animals, with widespread applications in cell biology.
authors.affiliation: Cancer Research UK Beatson Institute; Glasgow, UK.
authors.forename: Andreas K
authors.initials: AK
authors.lastname: Hock
authors.affiliation: Cancer Research UK Beatson Institute; Glasgow, UK.
authors.forename: Pearl
authors.initials: P
authors.lastname: Lee
authors.affiliation: Cancer Research UK Beatson Institute; Glasgow, UK.
authors.forename: Oliver Dk
authors.initials: OD
authors.lastname: Maddocks
authors.affiliation: Cancer Research UK Beatson Institute; Glasgow, UK.
authors.forename: Susan M
authors.initials: SM
authors.lastname: Mason
authors.affiliation: Cancer Research UK Beatson Institute; Glasgow, UK.
authors.forename: Karen
authors.initials: K
authors.lastname: Blyth
authors.affiliation: Cancer Research UK Beatson Institute; Glasgow, UK.
authors.forename: Karen H
authors.initials: KH
authors.lastname: Vousden
mesh_terms: D016475:3T3 Cells; D000818:Animals; D014408:Biomarkers, Tumor / Q000378:metabolism*; D002452:Cell Count; D002461:Cell Line, Transformed; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D005260:Female; D051858:Forkhead Transcription Factors / Q000235:genetics; D064593:Heterografts; D057166:High-Throughput Screening Assays; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D009374:Neoplasms, Experimental / Q000378:metabolism / Q000473:pathology*; D019265:Spectroscopy, Near-Infrared
chemical_list: D014408:Biomarkers, Tumor; D051858:Forkhead Transcription Factors; D008164:Luminescent Proteins; C090140:Whn protein
keywords: cancer; iRFP cell number quantification; in vivo; near-infrared fluorescence