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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.
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Total Internal Reflection Fluorescence Microscopy01:05

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Total internal reflection fluorescence microscopy or TIRF is an advanced microscopic technique used to visualize fluorophores in samples close to a solid surface with a higher refractive index, such as a glass coverslip. TIRF only allows fluorophores in proximity to the solid surface to be excited. When light from a medium with a lower refractive index (such as air) hits the glass coverslip at a critical angle, the light undergoes total internal reflection stead of passing through the glass.
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Updated: Sep 6, 2025

Rejection of Fluorescence Background in Resonance and Spontaneous Raman Microspectroscopy
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Bessel-beam illumination Raman microscopy.

Kazuki Bando1, Shumpei Yabuuchi1, Menglu Li1,2

  • 1Department of Applied Physics, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan.

Biomedical Optics Express
|July 5, 2022
PubMed
Summary
This summary is machine-generated.

Bessel beams enhance Raman imaging for label-free analysis of cell spheroids. This method improves contrast and reduces background in thick biological samples, enabling clearer intracellular molecule observation.

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Area of Science:

  • Biophotonics
  • Spectroscopy
  • Cellular Imaging

Background:

  • Raman imaging offers label-free analysis of biological samples.
  • Conventional methods struggle with background noise and low contrast in thick specimens.
  • Intracellular molecular analysis in live cell spheroids remains challenging.

Purpose of the Study:

  • To demonstrate Bessel beams for side-illumination slit-scanning Raman imaging.
  • To improve image contrast and background elimination for thick biological samples.
  • To enable label-free hyperspectral analysis of intracellular molecules in cell spheroids.

Main Methods:

  • Utilized Bessel beams for side illumination in slit-scanning Raman microscopy.
  • Employed aberration-resistant Bessel beams to enhance imaging.
  • Performed live cell spheroid imaging for comparative analysis.

Main Results:

  • Bessel beam side illumination significantly improved image contrast in Raman observations.
  • Effective background elimination was achieved, crucial for thick samples.
  • Demonstrated label-free investigation of intracellular molecules.

Conclusions:

  • Bessel beams combined with side illumination offer a superior approach for Raman imaging of cell spheroids.
  • The technique enhances clarity and reduces background noise compared to conventional epi-illumination.
  • This advancement facilitates detailed, label-free study of molecular distributions within live cells.