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Parallelized shifted-excitation Raman difference spectroscopy for fluorescence rejection in a temporary varying system.

ABSTRACT: A fluorescence background is one of the common interference factors of the Raman spectroscopic analysis in the biology field. Shifted-excitation Raman difference spectroscopy (SERDS), in which a slow (typically 1?Hz) modulation to excitation wavelength is coupled with a sequential acquisition of alternating shifted-excitation spectra, has been used to separate Raman scattering from excitation-shift insensitive background. This sequential method is susceptible to spectral change and thus is limited only to stable samples. We incorporated a fast laser modulation (200?Hz) and a mechanical streak camera into SERDS to effectively parallelize the SERDS measurement in a single exposure. The developed system expands the scope of SERDS to include temporary varying system. The proof of concept is demonstrated using highly fluorescent samples, including living algae. Quantitative performance in fluorescence rejection and the robustness of the method to the dynamic spectral change during the measurement are manifested.

SUBMITTER: Shimada R

PROVIDER: S-EPMC7065630 | biostudies-literature | 2019 Dec

REPOSITORIES: biostudies-literature

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Parallelized shifted-excitation Raman difference spectroscopy for fluorescence rejection in a temporary varying system.

Shimada Rintaro R Nakamura Takashi T Ozawa Takeaki T

Journal of biophotonics 20190828 12

A fluorescence background is one of the common interference factors of the Raman spectroscopic analysis in the biology field. Shifted-excitation Raman difference spectroscopy (SERDS), in which a slow (typically 1 Hz) modulation to excitation wavelength is coupled with a sequential acquisition of alternating shifted-excitation spectra, has been used to separate Raman scattering from excitation-shift insensitive background. This sequential method is susceptible to spectral change and thus is limit ...[more]

PMID: 31407507

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Project description:Optical fiber probe-based Raman spectroscopy systems are widely used for in situ measurements ranging from material characterization to biomedical applications. However, small Raman cross sections necessitate the use of high-power lasers or long exposure times that limit Raman's larger application to multiple research fields. This limitation can be overcome by collecting more Raman photons through additional collection fibers with taller detectors. This system configuration requires replacement of the detector and modification of the spectrograph to incorporate larger optical components, making it a costly and cumbersome option. In probe-based Raman systems, a typical detector image shows stacked collection fibers on the vertical axis and Raman spectra on the horizontal axis. While the vertical pixels are fully packed with multiple collection fibers, horizontal pixels have broad silent regions due to the narrow bandwidth of Raman peaks, potentially wasting valuable detector pixels. Here, we propose a new approach utilizing horizontally shifted collection fibers rather than vertically stacked ones. We designed and fabricated a novel collection fiber bundle that has horizontally shifted optical fibers in two vertical lines at the spectrograph entrance. This custom-made fiber bundle was incorporated into the imaging spectrograph to provide multiple horizontally shifted spectra on the detector. Through deconvolution, the original spectra can be recovered with an improved detection limit from greater photon collection. We demonstrate an enhanced limit of detection on various bioanalytes, such as glucose, urea, and lactate. Further, we applied the probe to measure tissue Raman spectra and successfully decomposed them into basis spectra, demonstrating the potential application of high-throughput in vivo tissue diagnosis. Our approach provides a simple, cost-effective, and universal method to increase the throughput without modifying existing Raman spectrometers.

Dataset Information

Parallelized shifted-excitation Raman difference spectroscopy for fluorescence rejection in a temporary varying system.

Publications

Parallelized shifted-excitation Raman difference spectroscopy for fluorescence rejection in a temporary varying system.

Similar Datasets

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