Structured line illumination Raman microscopy

Abstract

In the last couple of decades, the spatial resolution in optical microscopy has increased to unprecedented levels by exploiting the fluorescence properties of the probe. At about the same time, Raman imaging techniques have emerged as a way to image inherent chemical information in a sample without using fluorescent probes. However, in many applications, the achievable resolution is limited to about half the wavelength of excitation light. Here we report the use of structured illumination to increase the spatial resolution of label-free spontaneous Raman microscopy, generating highly detailed spatial contrast from the ensemble of molecular information in the sample. Using structured line illumination in slit-scanning Raman microscopy, we demonstrate a marked improvement in spatial resolution and show the applicability to a range of samples, including both biological and inorganic chemical component mapping. This technique is expected to contribute towards greater understanding of chemical component distributions in organic and inorganic materials.

Raman spectroscopy: techniques and applications in the life sciences

“…More recently structured illumination Raman microscopy has been reported to increase the spatial resolution [137].…”

Optimal 2D-SIM reconstruction by two filtering steps with Richardson-Lucy deconvolution

“…The apodization function has evolved into a parametrized function whose shape is empirically tuned to enhance the reconstruction and eliminate artifacts. In this sense, the reconstruction has become reliant on the appointed apodization function and is still a heavily debated issue23456789. In our method, we perform two filtering steps with the Richardson-Lucy (RL) deconvolution.…”

Three-dimensional wide-field pump-probe structured illumination microscopy

“…As an alternative method to point scanning, there have been publications to implement wide-field pump-probe microscopy based on coherent anti-Stokes Raman scattering (CARS) [19][20][21]. People have also applied SIM to pump-probe microscopy to increase the lateral resolution in two dimensions in various configurations including point scanning, line scanning and wide-field [22][23][24][25]. In this article, we extend these previous works to three dimensions (3D) in widefield configuration and propose a scheme that is compatible with pump-probe modalities where signal wavelength is not shifted from those of input beams (for example, signal wavelength is anti-Stokes shifted for CARS, but it is not for SRS).…”