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Implementation of a Nonlinear Microscope Based on Stimulated Raman Scattering
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Label-Free Biomedical Imaging Using High-Speed Lock-In Pixel Sensor for Stimulated Raman Scattering.

Kamel Mars1, De Xing Lioe2, Shoji Kawahito3

  • 1Research Institute of Electronics, Shizuoka University, 3-5-1 Johoku, Nakaku, Hamamatsu, Shizuoka 432-8011, Japan. kamel@idl.rie.shizuoka.ac.jp.

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PubMed
Summary
This summary is machine-generated.

Researchers developed a new CMOS image sensor for stimulated Raman scattering (SRS) microscopy. This detector enables faster, label-free biological imaging by efficiently capturing weak SRS signals at high speeds.

Keywords:
CMOS image sensorRaman shifthigh-speed modulationlock-in pixellow-noisestimulated Raman scattering

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Area of Science:

  • Biophotonics and Imaging
  • Materials Science and Engineering
  • Spectroscopy

Background:

  • Raman imaging offers label-free visualization of biological samples, crucial for understanding cellular processes without staining.
  • Stimulated Raman Scattering (SRS) microscopy has advanced rapidly, enabling high-speed and hyperspectral imaging.
  • A key limitation in SRS microscopy is the lack of suitable detectors capable of MHz modulation rates for parallel detection of weak signals while rejecting strong background light.

Purpose of the Study:

  • To develop a novel complementary metal-oxide semiconductor (CMOS) image sensor with high-speed lock-in pixels specifically designed for stimulated Raman scattering (SRS) applications.
  • To address the challenge of detecting weak SRS signals at MHz modulation frequencies while effectively eliminating strong background laser light.
  • To enable faster and more efficient hyperspectral and multi-focus SRS imaging of biological samples.

Main Methods:

  • A custom CMOS image sensor was designed featuring high-speed lock-in pixels capable of in-pixel signal differencing (Stokes-on vs. Stokes-off) at 20 MHz.
  • An integrated lateral electric field charge modulator (LEFM) with two-step ion implantation was employed for efficient extraction and amplification of small SRS signals.
  • The pixel architecture includes low-pass filters, sample-and-hold circuits, and switched capacitor integrators with fully differential amplifiers, fabricated using 0.11 μm CMOS technology.

Main Results:

  • The prototype CMOS image sensor successfully acquired SRS spectra and images of stearic acid and 3T3-L1 cell samples.
  • The sensor demonstrated the capability to extract and amplify weak SRS signals at a 20 MHz modulation frequency directly within the pixel.
  • The results validate the potential for high-speed, label-free SRS imaging, including hyperspectral and multi-focus capabilities.

Conclusions:

  • The developed CMOS image sensor with high-speed lock-in pixels effectively overcomes detector limitations in SRS microscopy.
  • This technology paves the way for real-time, label-free hyperspectral and multi-focus SRS imaging at video rates.
  • Further modifications to the pixel architecture and acquisition system could enhance performance for advanced biological imaging applications.