Harmonic holography for three-dimensional cellular imaging

Rachel Grange, Chia-Lung Hsieh, Demetri Psaltis, Ye Pu
2009
Conference paper

Abstract

Luminescent markers play a key role in imaging techniques for life sciences since they provide a contrast mechanism between signal and background. We describe a new type of marker using second harmonic generation (SHG) from noncentrosymmetric BaTiO3 nanocrystals. These nanoparticles are attractive due to their stable, non-saturating and coherent signal with a femtosecond-scale response time and a broad flexibility in the choice of excitation wavelength. We use the coherent SHG signal from BaTiO3 nanoparticles for three-dimensional (3D) imaging without scanning. We built a harmonic holographic (H2) microscope which records digital holograms at the second harmonic frequency. Highresolution 3D distributions of these SHG markers in mammalian cells are successfully captured and interpreted by the H2 microscope. © 2009 SPIE.

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https://infoscience.epfl.ch/record/155335?ln=en

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Three-dimensional harmonic holographic microcopy using nanoparticles as probes for cell imaging

Rachel Grange, Chia-Lung Hsieh, Demetri Psaltis, Ye Pu

Luminescent markers play a key role in imaging techniques for life science since they provide a contrast mechanism between signal and background. We describe a new type of marker using second harmonic generation (SHG) from noncentrosymmetric BaTiO3 nanocrystals. These nanoparticles are attractive due to their stable, non-saturating and coherent signal with a femtosecond-scale response time and broad flexibility in the choice of excitation wavelength. We obtained monodispersed BaTiO3 nanoparticles in colloidal suspensions by coating the particle surface with amine groups. We characterized the SHG efficiency of 90-nm BaTiO3 particles experimentally and theoretically. Moreover, we use the coherent SHG signal from BaTiO3 nanoparticles for three-dimensional (3D) imaging without scanning. We built a harmonic holographic ( H-2) microscope which records digital holograms at the second harmonic frequency. For the first time, high-resolution 3D distributions of these SHG markers in mammalian cells are successfully captured and interpreted by the H-2 microscope. (c) 2009 Optical Society of America

Optical Society of America2009

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We develop second-harmonic nanoparticles as the contrast agents for cell imaging. Second-harmonic nanoparticles show promise as cell imaging probes due to their non-bleaching, non-blinking, and coherent signal. Nanoparticles of noncentrosymmetric crystal structures have high second-harmonic generation (SHG) efficiency and provide high contrast in a generally non-structured cell environment. Here, we use barium titanate (BaTiO3) nanoparticles with tetragonal crystal structure as imaging probes. Cytotoxicity tests performed on BaTiO3 nanoparticles with mammalian cells did not result in toxic effects. Specifically we observed no change in the cell metabolism after 24 hours incubation of the cells with high concentration of BaTiO3 nanoparticles. We demonstrate two methods of cell labeling with BaTiO 3 nanoparticles for imaging. One is non-specific labeling via endocytosis of the cells, which results in a great number of the nanoparticles randomly distributed inside the cells. The other is specific labeling via surface functionalization of the nanoparticles with antibodies, which enables us to label specific cell membrane proteins with the nanoparticles. SHG imaging is compatible to two-photon microscopy and the SHG signal from nanoparticles can be easily detected with a standard two-photon confocal microscope. Our work provides the opportunity for long-term, three-dimensional cell tracking with secondharmonic nanoparticles. © 2010 SPIE.

Spie-Int Soc Optical Engineering, Po Box 10, Bellingham, Wa 98227-0010 Usa2010

Barium titanate nanoparticles used as second harmonic radiation imaging probes for cell imaging

Rachel Grange, Chia-Lung Hsieh, Demetri Psaltis, Ye Pu

Luminescent markers play a key role in imaging techniques for life science since they provide a contrast mechanism between signal and background. We describe a new type of marker using second harmonic generation (SHG) from noncentrosymmetric BaTiO3 nanocrystals. These nanoparticles are attractive due to their stable, non-saturating and coherent signal with a femtosecond-scale response time and broad flexibility in the choice of excitation wavelength. In this paper, we report the use of nanoparticles for cell imaging. We first stabilized the BaTiO3 nanoparticles in suspension and characterized the optical properties. We also demonstrated the functionalization of BaTiO3 nanoparticles by conjugating IgG antibody on the surface of the nanoparticles. These functionalized nanocrystals are capable of specific labeling the antigens of interest. © 2009 SPIE-OSA.

2009