In vivo cell tracking and quantification method in adult zebrafish

Li Zhang; Clemens Alt; Pulin Li; Richard M. White; Leonard I. Zon; Xunbin Wei; Charles P. Lin

doi:10.1117/12.909632

9 February 2012 In vivo cell tracking and quantification method in adult zebrafish

Li Zhang, Clemens Alt, Pulin Li, Richard M. White, Leonard I. Zon, Xunbin Wei, Charles P. Lin

Proceedings Volume 8225, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues X; 82250P (2012) https://doi.org/10.1117/12.909632
Event: SPIE BiOS, 2012, San Francisco, California, United States

Abstract

Zebrafish have become a powerful vertebrate model organism for drug discovery, cancer and stem cell research. A recently developed transparent adult zebrafish using double pigmentation mutant, called casper, provide unparalleled imaging power in in vivo longitudinal analysis of biological processes at an anatomic resolution not readily achievable in murine or other systems. In this paper we introduce an optical method for simultaneous visualization and cell quantification, which combines the laser scanning confocal microscopy (LSCM) and the in vivo flow cytometry (IVFC). The system is designed specifically for non-invasive tracking of both stationary and circulating cells in adult zebrafish casper, under physiological conditions in the same fish over time. The confocal imaging part in this system serves the dual purposes of imaging fish tissue microstructure and a 3D navigation tool to locate a suitable vessel for circulating cell counting. The multi-color, multi-channel instrument allows the detection of multiple cell populations or different tissues or organs simultaneously. We demonstrate initial testing of this novel instrument by imaging vasculature and tracking circulating cells in CD41: GFP/Gata1: DsRed transgenic casper fish whose thrombocytes/erythrocytes express the green and red fluorescent proteins. Circulating fluorescent cell incidents were recorded and counted repeatedly over time and in different types of vessels. Great application opportunities in cancer and stem cell researches are discussed.

Citation Download Citation

Li Zhang, Clemens Alt, Pulin Li, Richard M. White, Leonard I. Zon, Xunbin Wei, and Charles P. Lin "In vivo cell tracking and quantification method in adult zebrafish", Proc. SPIE 8225, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues X, 82250P (9 February 2012); https://doi.org/10.1117/12.909632

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KEYWORDS

In vivo imaging

Confocal microscopy

Green fluorescent protein

Imaging systems

Flow cytometry

Stem cells

Blood vessels

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