Label-Free Biomedical Imaging Using High-Speed Lock-In Pixel Sensor for Stimulated Raman Scattering.

Kamel Mars,Keita Yasutomi,Takahiro Yamada,De Xing Lioe,Shoji Kawahito,Keiichiro Kagawa,Mamoru Hashimoto

doi:10.3390/s17112581

Abstract

Raman imaging eliminates the need for staining procedures, providing label-free imaging to study biological samples. Recent developments in stimulated Raman scattering (SRS) have achieved fast acquisition speed and hyperspectral imaging. However, there has been a problem of lack of detectors suitable for MHz modulation rate parallel detection, detecting multiple small SRS signals while eliminating extremely strong offset due to direct laser light. In this paper, we present a complementary metal-oxide semiconductor (CMOS) image sensor using high-speed lock-in pixels for stimulated Raman scattering that is capable of obtaining the difference of Stokes-on and Stokes-off signal at modulation frequency of 20 MHz in the pixel before reading out. The generated small SRS signal is extracted and amplified in a pixel using a high-speed and large area lateral electric field charge modulator (LEFM) employing two-step ion implantation and an in-pixel pair of low-pass filter, a sample and hold circuit and a switched capacitor integrator using a fully differential amplifier. A prototype chip is fabricated using 0.11 μm CMOS image sensor technology process. SRS spectra and images of stearic acid and 3T3-L1 samples are successfully obtained. The outcomes suggest that hyperspectral and multi-focus SRS imaging at video rate is viable after slight modifications to the pixel architecture and the acquisition system.

Highlights

Coherent Raman scattering (CRS) is gaining popularity to provide label-free biomedical imaging.The CRS finds its applications in a wide range of biology and medical imaging [1,2,3,4,5,6,7,8], in which the use of exogenous dye will perturb the molecules of interest
To address the limitation in noise caused by the use of sub-pixels array to achieve a large area detector to obtain a better output signal stability and to improve the signal-to-noise ratio (SNR), we propose a new structure of pinned-photodiode-based detector
The power of the pump laser and the Stokes laser is set to 3.5 mW and 4.5 mW, respectively, and the total integration cycles are set to 700 cycles

Summary

Introduction

The CRS finds its applications in a wide range of biology and medical imaging [1,2,3,4,5,6,7,8], in which the use of exogenous dye will perturb the molecules of interest. Another advantage of the CRS is the high sensitivity that overcomes the drawback of low-speed imaging by spontaneous Raman scattering. Both CARS and SRS excite the sample with a pair of lasers, namely the pump at frequency ωp and the Stokes at frequency ωs .

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Conclusion