Ms. Marie-Charlotte Chandeclerc Profile

Marie-Charlotte Chandeclerc

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Proceedings Article | 30 May 2022 Presentation

Holography-based 3D real-time single-particle tracking for intraneuronal transport characterization

Florian Semmer, Marie-Charlotte Chandeclerc, François Treussart, Karen Perronet, François Marquier

Proceedings Volume PC12144, PC1214408 (2022) https://doi.org/10.1117/12.2622036

KEYWORDS: Holography, Nonlinear optics, Digital micromirror devices, Two photon excitation microscopy, Spatial resolution, Neurons, Nanoparticles, Nanocrystals, Multiphoton microscopy, Microscopy

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Neurodegenerative diseases such as Alzheimer’s disease present abnormalities in intraneuronal transport, suggesting the relevance of measuring this key biological process. In 2017, a sensitive method to measure changes in intraneuronal endosomal transport has been reported in 2D cultures of neurons using fluorescent nanodiamonds (fNDs) [1]. The high brightness, photostability and absence of cytotoxicity allow fNDs to be tracked with 50 nm spatial and 50 ms time resolutions. This nanoparticle tracking based-approach applies also to multiphoton imaging, opening the possibility of transport measurement in vivo. We use nanocrystals possessing a large nonlinear second order optical response. First results indicate that the intraneuronal transport measurement can be inferred from nonlinear microscopy data, opening applications to thicker samples owing to the low background of multiphoton imaging. In order to get a high spatio-temporal resolution (around 10 nanometers at 1 ms), we are developing a two-photon microscope, based on a digital holography method [2]. A Digital Micromirror Device (DMD) is used as a spatial light modulator, allowing a fast 3D motion of the excitation volume. We aim at reaching a time resolution below the millisecond and super-localization regime in the tens nanometer range using orbital tracking. References : [1] S. Haziza, et al. Nat. Nanotechnol. 12 (2017), 322. [2] Geng, Q., Gu, C., Cheng, J. & Chen, S. Digital micromirror device-based two-photon microscopy for three-dimensional and random-access imaging. Optica 4, 674 (2017)

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