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Proceedings Paper

In vivo cell tracking and quantification method in adult zebrafish
Author(s): Li Zhang; Clemens Alt; Pulin Li; Richard M. White; Leonard I. Zon; Xunbin Wei; Charles P. Lin
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Paper 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.

Paper Details

Date Published: 9 February 2012
PDF: 9 pages
Proc. SPIE 8225, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues X, 82250P (9 February 2012); doi: 10.1117/12.909632
Show Author Affiliations
Li Zhang, Massachusetts General Hospital, Harvard Medical School (United States)
Fudan Univ. (China)
Clemens Alt, Massachusetts General Hospital, Harvard Medical School (United States)
Pulin Li, Harvard Medical School (United States)
Richard M. White, Harvard Medical School (United States)
Leonard I. Zon, Harvard Medical School (United States)
Xunbin Wei, Shanghai Jiaotong Univ. (China)
Fudan Univ. (China)
Charles P. Lin, Massachusetts General Hospital, Harvard Medical School (United States)

Published in SPIE Proceedings Vol. 8225:
Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues X
Daniel L. Farkas; Dan V. Nicolau; Robert C. Leif, Editor(s)

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