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

Fingerprint

Rheology
embryos
Fluorescence Spectrometry
Endothelial cells
Blood vessels
Zebrafish
Lighting
Charge coupled devices
Velocity measurement
Transparency
Blood Proteins
Blood
Embryonic Structures
Photons
Fluorescence
illumination
Cells
Spectroscopy
Fluxes
Plasmas

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

Cite this

Pozzi, P., Rossetti, L., Sironi, L., Freddi, S., D'Alfonso, L., Caccia, M., ... Giuseppe, C. (2013). Structured illumination fluorescence correlation spectroscopy for velocimetry in zebrafish embryos. In Progress in Biomedical Optics and Imaging - Proceedings of SPIE (Vol. 8580). [85800V] https://doi.org/10.1117/12.2002082

Structured illumination fluorescence correlation spectroscopy for velocimetry in zebrafish embryos. / Pozzi, Paolo; Rossetti, Leone; Sironi, Laura; Freddi, Stefano; D'Alfonso, Laura; Caccia, Michele; Bouzin, Margaux; Collini, Maddalena; Giuseppe, Chirico.

Progress in Biomedical Optics and Imaging - Proceedings of SPIE. Vol. 8580 2013. 85800V.

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

Pozzi, P, Rossetti, L, Sironi, L, Freddi, S, D'Alfonso, L, Caccia, M, Bouzin, M, Collini, M & Giuseppe, C 2013, Structured illumination fluorescence correlation spectroscopy for velocimetry in zebrafish embryos. in Progress in Biomedical Optics and Imaging - Proceedings of SPIE. vol. 8580, 85800V, Dynamics and Fluctuations in Biomedical Photonics VIII Conference, San Francisco, CA, United States, 2/2/13. https://doi.org/10.1117/12.2002082
Pozzi P, Rossetti L, Sironi L, Freddi S, D'Alfonso L, Caccia M et al. Structured illumination fluorescence correlation spectroscopy for velocimetry in zebrafish embryos. In Progress in Biomedical Optics and Imaging - Proceedings of SPIE. Vol. 8580. 2013. 85800V https://doi.org/10.1117/12.2002082
Pozzi, Paolo ; Rossetti, Leone ; Sironi, Laura ; Freddi, Stefano ; D'Alfonso, Laura ; Caccia, Michele ; Bouzin, Margaux ; Collini, Maddalena ; Giuseppe, Chirico. / Structured illumination fluorescence correlation spectroscopy for velocimetry in zebrafish embryos. Progress in Biomedical Optics and Imaging - Proceedings of SPIE. Vol. 8580 2013.
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