Parallelized shifted‐excitation Raman difference spectroscopy for fluorescence rejection in a temporary varying system
| Autor: | Takashi Nakamura, Takeaki Ozawa, Rintaro Shimada |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Materials science
SERDS General Physics and Astronomy Spectrum Analysis Raman 01 natural sciences Fluorescence General Biochemistry Genetics and Molecular Biology Spectral line 010309 optics symbols.namesake Optics Full Article 0103 physical sciences General Materials Science Spectroscopy business.industry Streak camera Chlamydomonas Full Articles 010401 analytical chemistry General Engineering General Chemistry mechanical streak camera Biomechanical Phenomena 0104 chemical sciences multivariate analysis Biology (field) Calibration Raman spectroscopy symbols business background correction Raman scattering Excitation |
| Zdroj: | Journal of Biophotonics |
| ISSN: | 1864-0648 1864-063X |
| DOI: | 10.1002/jbio.201960028 |
| Popis: | 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. We present parallelized shifted excitation Raman difference spectroscopy by the synchronized operation of a mechanical streak camera with a wavelength tunable light source. The parallel detection scheme facilitates the effective and accurate fluorescence rejection from a temporary varying system having faster dynamics than the spectral acquisition rate. Quantitative performance in fluorescence rejection and the robustness of the method to the dynamic spectral change during the measurement are manifested. |
| Databáze: | OpenAIRE |
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