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20 June 2014 Electron multiplying charge-coupled device-based fluorescence cross-correlation spectroscopy for blood velocimetry on zebrafish embryos
Paolo Pozzi, Laura Sironi, Laura D’Alfonso, Margaux Bouzin, Maddalena Collini, Giuseppe Chirico, Piersandro Pallavicini, Franco Cotelli, Efrem A. Foglia
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Journal of Biomedical Optics, Vol. 19, Issue 6, 067007 (June 2014). https://doi.org/10.1117/1.JBO.19.6.067007
Abstract
Biomedical issues in vasculogenesis and cardiogenesis require methods to follow hemodynamics with high spatial (micrometers) and time (milliseconds) resolution. At the same time, we need to follow relevant morphogenetic processes on large fields of view. Fluorescence cross-correlation spectroscopy coupled to scanning or wide-field microscopy meets these needs but has limited flexibility in the excitation pattern. To overcome this limitation, we develop here a two-photon two-spots setup coupled to an all-reflective near-infrared (NIR) optimized scanning system and to an electron multiplying charge-coupled device. Two NIR laser spots are spaced at adjustable micron-size distances (1 to 50 μm) by means of a Twyman-Green interferometer and repeatedly scanned on the sample, allowing acquisition of information on flows at 4 ms–3 μm time-space resolution in parallel on an extended field of view. We analyze the effect of nonhomogeneous and variable flow on the cross-correlation function by numerical simulations and show exemplary application of this setup in studies of blood flow in zebrafish embryos in vivo. By coupling the interferometer with the scanning mirrors and by computing the cross-correlation function of fluorescent red blood cells, we are able to map speed patterns in embryos’ vessels.
© 2014 Society of Photo-Optical Instrumentation Engineers (SPIE) 0091-3286/2014/$25.00 © 2014 SPIE
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Paolo Pozzi, Laura Sironi, Laura D’Alfonso, Margaux Bouzin, Maddalena Collini, Giuseppe Chirico, Piersandro Pallavicini, Franco Cotelli, and Efrem A. Foglia "Electron multiplying charge-coupled device-based fluorescence cross-correlation spectroscopy for blood velocimetry on zebrafish embryos," Journal of Biomedical Optics 19(6), 067007 (20 June 2014). https://doi.org/10.1117/1.JBO.19.6.067007
Published: 20 June 2014
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Cited by 10 scholarly publications.
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KEYWORDS
Diffusion
Capillaries
Luminescence
Blood
Veins
Blood circulation
Solids
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