In this visualized experiment, protocol details are provided for in vitro labeling of human embryonic stem cells (hESC) with second harmonic generation nanoparticles (HNPs). The latter are a new family of probes recently introduced for labeling biological samples for multi-photon imaging. HNPs are capable of doubling the frequency of excitation light by the nonlinear optical process of second harmonic generation with no restriction on the excitation wavelength.Multi-photon based methodologies for hESC differentiation into cardiac clusters (maintained as long term air-liquid cultures) are presented in detail. In particular, evidence on how to maximize the intense second harmonic (SH) emission of isolated HNPs during 3D monitoring of beating cardiac tissue in 3D is shown. The analysis of the resulting images to retrieve 3D displacement patterns is also detailed.

Harmonic nanoparticles for regenerative research

Ronzoni, Flavio;
2014

Abstract

In this visualized experiment, protocol details are provided for in vitro labeling of human embryonic stem cells (hESC) with second harmonic generation nanoparticles (HNPs). The latter are a new family of probes recently introduced for labeling biological samples for multi-photon imaging. HNPs are capable of doubling the frequency of excitation light by the nonlinear optical process of second harmonic generation with no restriction on the excitation wavelength.Multi-photon based methodologies for hESC differentiation into cardiac clusters (maintained as long term air-liquid cultures) are presented in detail. In particular, evidence on how to maximize the intense second harmonic (SH) emission of isolated HNPs during 3D monitoring of beating cardiac tissue in 3D is shown. The analysis of the resulting images to retrieve 3D displacement patterns is also detailed.
Bioengineering
Issue 87
multi-photon imaging
human embryonic stem cells (ESC)
nanoparticles
embryoid bodies (EBs)
cardiomyocyte differentiation
cardiac contraction
air-liquid cultures
Cell Communication
Cell Differentiation
Embryonic Stem Cells
Humans
Imaging, Three-Dimensional
Microscopy, Fluorescence, Multiphoton
Myocytes, Cardiac
Nanoparticles
Niobium
Oxides
Potassium
Regeneration
Regenerative Medicine
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11699/62155
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