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Related Concept Videos

Raman Spectroscopy Instrumentation: Overview01:26

Raman Spectroscopy Instrumentation: Overview

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A conventional Raman spectrophotometer includes a laser source, a sample holding system, a wavelength selector, and a detector.
The monochromatic laser source, typically using visible or near-infrared radiation, generates a highly focused beam of light. This light interacts with the molecules of the sample, scattering some of the light. Liquid and gaseous samples are usually tested in ordinary glass capillaries, while solids can be analyzed as powders packed in capillaries or as potassium...
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Raman Spectroscopy: Overview01:20

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The underlying principle of Raman spectroscopy is based on the interaction between light and matter, specifically molecules' inelastic scattering of photons. When a monochromatic beam of light, typically from a laser source, interacts with a sample, most scattered light has the same frequency as the incident light. This is known as Rayleigh scattering.
However, a small fraction of the scattered light exhibits a frequency shift due to the exchange of energy between the incident photons and...
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Super-resolution Fluorescence Microscopy01:37

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Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been...
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Ultrafast Time-resolved Near-IR Stimulated Raman Measurements of Functional π-conjugate Systems
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Ultrafast Simultaneous Raman-Fluorescence Spectroscopy.

Matthew Lindley1, Kotaro Hiramatsu1,2,3, Hayate Nomoto1

  • 1Department of Chemistry , The University of Tokyo , Tokyo 113-0033 , Japan.

Analytical Chemistry
|November 28, 2019
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Summary
This summary is machine-generated.

Scientists developed a fast, multimodal spectrometer combining Raman and fluorescence spectroscopy for bioanalysis. This new fluoRaman technique enables high-throughput single-cell analysis, overcoming previous signal interference issues.

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Area of Science:

  • Bioanalytical Chemistry
  • Spectroscopy
  • Microbiology

Background:

  • Raman and fluorescence spectroscopy are complementary bioanalytical techniques.
  • Fluorescence signals often overwhelm weak Raman signals, limiting combined use.
  • Existing methods struggle with simultaneous, high-speed Raman and fluorescence measurements.

Purpose of the Study:

  • To develop a high-speed, multimodal spectrometer for simultaneous Raman and fluorescence measurements.
  • To overcome signal interference challenges in combined spectroscopic techniques.
  • To enable rapid, label-free and targeted analysis in microbiological assays.

Main Methods:

  • Utilized Fourier-transform-coherent anti-Stokes Raman scattering (FT-CARS) and Fourier-transform-two-photon excitation (FT-TPE).
  • Employed a femtosecond pulse laser coupled to a rapid-scan Michelson interferometer.
  • Achieved high acquisition speeds of 24,000 spectra per second.

Main Results:

  • Successfully demonstrated simultaneous, high-speed Raman and fluorescence measurements.
  • Validated the system by measuring coumarin dyes in organic solvents.
  • Applied the technique to fluoRaman flow cytometry of Haematococcus pluvialis cells at ~10 events/sec.

Conclusions:

  • The developed ultrafast fluoRaman spectrometer overcomes previous limitations in combining Raman and fluorescence spectroscopy.
  • This technology enables high-throughput, single-cell analysis for studying cellular responses.
  • Potential applications include large-scale metabolic stress response analysis in microbial populations.