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Measurement of Scattering Nonlinearities from a Single Plasmonic Nanoparticle
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Simultaneous scattering compensation at multiple points in multi-photon microscopy.

Molly A May1, Kai K Kummer2, Marie-Luise Edenhofer2

  • 1Institute of Biomedical Physics, Medical University of Innsbruck, Müllerstraße 44, 6020 Innsbruck, Austria.

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|January 10, 2022
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Summary
This summary is machine-generated.

This study enhances two-photon fluorescence imaging depth by using Dynamic Adaptive Scattering compensation Holography (DASH) with a liquid crystal spatial light modulator. This method effectively corrects scattering in living mouse tissue, improving imaging capabilities.

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Area of Science:

  • Biomedical Optics
  • Neuroscience Imaging
  • Advanced Microscopy Techniques

Background:

  • Two-photon fluorescence imaging depth is limited by biological tissue scattering.
  • Current wavefront correction methods are effective only over small sample regions (1–10 µm).
  • Sample conjugate correction geometries can expand the field of view in planar scattering samples.

Purpose of the Study:

  • To improve the field of view and correction efficiency for scattering in two-photon fluorescence imaging.
  • To apply a novel sensor-less scattering correction scheme, Dynamic Adaptive Scattering compensation Holography (DASH), in a sample conjugate configuration.
  • To demonstrate simultaneous scattering correction at multiple field points using a high pixel count nematic liquid crystal spatial light modulator (LC-SLM).

Main Methods:

  • Implementation of the DASH algorithm with a large LC-SLM in a sample conjugate setup.
  • Simultaneous correction for scattering across multiple, distributed field points within the objective lens's field of view.
  • Measurement of correction times to assess feasibility for dynamic biological samples.

Main Results:

  • Achieved scattering correction times of approximately 10 seconds per field point, despite the LC-SLM's slow refresh rate.
  • Demonstrated effective scattering compensation at multiple sites within living mouse hippocampal tissue slices.
  • Showcased the capability of correcting scattering over a larger field of view compared to traditional methods.

Conclusions:

  • DASH, when implemented with a large LC-SLM in a sample conjugate configuration, significantly enhances two-photon fluorescence imaging depth and field of view.
  • The achieved correction speed is sufficient for counteracting scattering in dynamic biological samples like living mouse brain tissue.
  • This approach offers a promising method for deeper and wider field imaging in scattering biological tissues.