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Proceedings Paper

Structured illumination fluorescence correlation spectroscopy for velocimetry in Zebrafish embryos
Author(s): Paolo Pozzi; Leone Rossetti; Laura Sironi; Stefano Freddi; Laura D'Alfonso; Michele Caccia; Margaux Bouzin; Maddalena Collini; Giuseppe Chirico
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Paper Abstract

The vascular system of Zebrafish embryos is studied by means of Fluorescence Correlation and Image Correlation Spectroscopy. The long term project addresses biologically relevant issues concerning vasculogenesis and cardiogenesis and in particular mechanical interaction between blood flow and endothelial cells. To this purpose we use Zebrafish as a model system since the transparency of its embryos facilitates morphological observation of internal organs in-vivo. The correlation analysis provides quantitative characterization of fluxes in blood vessels in vivo. We have pursued and compared two complementary routes. In a first one we developed a two-spots two-photon setup in which the spots are spaced at adjustable micron-size distances (1-40 μm) along a vessel and the endogenous (autofluorescence) or exogenous (dsRed transgenic erythrocytes) signal is captured with an EM-CCD and cross-correlated. In this way we are able to follow the morphology of the Zebrafish embryo, simultaneously measure the heart pulsation, the velocity of red cells and of small plasma proteins. These data are compared to those obtained by image correlations on Zebrafish vessels. The two methods allows to characterize the motion of plasma fluids and erythrocytes in healthy Zebrafish embryos to be compared in the future to pathogenic ones.

Paper Details

Date Published: 1 March 2013
PDF: 9 pages
Proc. SPIE 8580, Dynamics and Fluctuations in Biomedical Photonics X, 85800V (1 March 2013); doi: 10.1117/12.2002082
Show Author Affiliations
Paolo Pozzi, Univ. degli Studi di Milano-Bicocca (Italy)
Leone Rossetti, Univ. degli Studi di Milano-Bicocca (Italy)
Laura Sironi, Univ. degli Studi di Milano-Bicocca (Italy)
Stefano Freddi, Univ. degli Studi di Milano-Bicocca (Italy)
Laura D'Alfonso, Univ. degli Studi di Milano-Bicocca (Italy)
Michele Caccia, Univ. degli Studi di Milano-Bicocca (Italy)
Margaux Bouzin, Univ. degli Studi di Milano-Bicocca (Italy)
Maddalena Collini, Univ. degli Studi di Milano-Bicocca (Italy)
Giuseppe Chirico, Univ. degli Studi di Milano-Bicocca (Italy)


Published in SPIE Proceedings Vol. 8580:
Dynamics and Fluctuations in Biomedical Photonics X
Valery V. Tuchin; Donald D. Duncan; Kirill V. Larin; Martin J. Leahy; Ruikang K. Wang, Editor(s)

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