Real-time dynamics of molecular order and orientation by fast polarization modulation in coherent Raman scattering microscopy (Conference Presentation)

Matthias Hofer; Naveen K. Balla; Sophie Brasselet

doi:10.1117/12.2509692

4 March 2019 Real-time dynamics of molecular order and orientation by fast polarization modulation in coherent Raman scattering microscopy (Conference Presentation)

Matthias Hofer, Naveen K. Balla, Sophie Brasselet

Author Affiliations +

Proceedings Volume 10889, High-Speed Biomedical Imaging and Spectroscopy IV; 1088909 (2019) https://doi.org/10.1117/12.2509692
Event: SPIE BiOS, 2019, San Francisco, California, United States

Abstract

Probing vibrational modes of molecules by Coherent Raman Scattering (CRS) provides a non-invasive, label free and chemically sensitive imaging modality that is of a large interest for the study of biological systems. Stimulated Raman Scattering and Coherent Anti-Stokes Raman Scattering provide quantitative imaging of the density of resonant vibrational modes within a focal volume. However, those modes are also sensitive to the incident polarization of light, which can give additional insight into the structural order and orientation of molecular assemblies. Accessing such sub-diffraction molecular order information opens ways to understand the interplay between organization and function in biological systems, as well as its alteration in the progression of diseases. A limiting factor of polarized-CRS imaging so far was the long acquisition times due to a slow step wise rotation of the incident polarization state, leading to image acquisition times of 100x100 μm field of views in the minute time scales. In this work, we use electro-optic modulation to fully rotate the incident polarization state at rates of 100 kHz. Coupled with lock-in detection, we show that one can obtain order and orientation parameters of molecular vibrations down to sub-seconds time scales per image. Additional incident intensity modulation permits to access simultaneously molecular density information, with no compromise in signal to noise conditions. Using this method, we demonstrate real-time deformation and dynamics of model membrane systems (multilamellar vesicles). Moreover, we show the sensitivity of the technique down to one single lipid bilayer as present in red blood cell ghosts.

Conference Presentation

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Matthias Hofer, Naveen K. Balla, and Sophie Brasselet "Real-time dynamics of molecular order and orientation by fast polarization modulation in coherent Raman scattering microscopy (Conference Presentation)", Proc. SPIE 10889, High-Speed Biomedical Imaging and Spectroscopy IV, 1088909 (4 March 2019); https://doi.org/10.1117/12.2509692

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KEYWORDS

Polarization

Modulation

Raman scattering

Microscopy

Imaging systems

Systems modeling

CARS tomography

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