Structured illumination fluorescence correlation spectroscopy for velocimetry in zebrafish embryos

Paolo Pozzi, Leone Rossetti, Laura Sironi, Stefano Freddi, Laura D'Alfonso, Michele Caccia, Margaux Bouzin, Maddalena Collini, Chirico Giuseppe

Research output: Chapter in Book/Report/Conference proceedingConference contribution

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.

Original languageEnglish
Title of host publicationProgress in Biomedical Optics and Imaging - Proceedings of SPIE
Volume8580
DOIs
Publication statusPublished - 2013
EventDynamics and Fluctuations in Biomedical Photonics VIII Conference - San Francisco, CA, United States
Duration: Feb 2 2013Feb 4 2013

Other

OtherDynamics and Fluctuations in Biomedical Photonics VIII Conference
CountryUnited States
CitySan Francisco, CA
Period2/2/132/4/13

Keywords

  • Fluorescence Correlation
  • Velocimetry
  • Zebrafish

ASJC Scopus subject areas

  • Atomic and Molecular Physics, and Optics
  • Electronic, Optical and Magnetic Materials
  • Biomaterials
  • Radiology Nuclear Medicine and imaging

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