title: An optical platform for cell tracking in adult zebrafish.
abstract: Adult zebrafish are being increasingly used as a model in cancer and stem cell research. Here we describe an integrated optical system that combines a laser scanning confocal microscope (LSCM) and an in vivo flow cytometer (IVFC) for simultaneous visualization and cell quantification. The system is set up specifically for non-invasive tracking of both stationary and circulating cells in adult zebrafish (casper) that have been engineered to be optically transparent. Confocal imaging in this instrument serves the dual purpose of visualizing fish tissue microstructure and an imaging-based guide to locate a suitable vessel for quantitative analysis of circulating cells by IVFC. We demonstrate initial testing of this novel instrument by imaging the transparent adult zebrafish casper vasculature and tracking circulating cells in CD41-GFP/Gata1-DsRed transgenic fish whose thrombocytes/erythrocytes express the green and red fluorescent proteins. In vivo measurements allow cells to be tracked under physiological conditions in the same fish over time, without drawing blood samples or sacrificing animals. We also discuss the potential applications of this instrument in biomedical research.
authors.affiliation: Advanced microscopy program, Center for Systems Biology and Wellman Center for Photomedicine, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts 02114, USA.
authors.forename: Li
authors.initials: L
authors.lastname: Zhang
authors.forename: Clemens
authors.initials: C
authors.lastname: Alt
authors.forename: Pulin
authors.initials: P
authors.lastname: Li
authors.forename: Richard M
authors.initials: RM
authors.lastname: White
authors.forename: Leonard I
authors.initials: LI
authors.lastname: Zon
authors.forename: Xunbin
authors.initials: X
authors.lastname: Wei
authors.forename: Charles P
authors.initials: CP
authors.lastname: Lin
mesh_terms: D000375:Aging / Q000502:physiology*; D000818:Animals; D030801:Animals, Genetically Modified; D001808:Blood Vessels / Q000166:cytology; D058948:Cell Tracking / Q000295:instrumentation* / Q000379:methods*; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism; D005434:Flow Cytometry; D049452:Green Fluorescent Proteins / Q000378:metabolism; D018613:Microscopy, Confocal; D055095:Optics and Photonics / Q000295:instrumentation*; D040201:Platelet Membrane Glycoprotein IIb / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D015203:Reproducibility of Results; D015027:Zebrafish / Q000235:genetics / Q000378:metabolism*
chemical_list: D040201:Platelet Membrane Glycoprotein IIb; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins