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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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IR Spectrometers01:25

IR Spectrometers

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There are two main infrared (IR) spectrophotometers: dispersive IR spectrometers and Fourier transform infrared (FTIR) spectrometers. In a dispersive IR spectrometer, a beam of infrared radiation produced by a hot wire is divided into two parallel equal-intensity beams using mirrors. One beam passes through the sample, while another is a reference beam. The beams then move through the monochromator, which separates the radiations into a continuous spectrum of different frequencies. The...
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Infrared (IR) Spectroscopy: Overview01:09

Infrared (IR) Spectroscopy: Overview

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When electromagnetic radiation passes through a material, atoms or molecules transition from a lower to a higher energy state by absorbing radiation corresponding to the energy difference between the two states. The absorption of infrared (IR) radiation causes transitions between vibrational energy levels in a molecule. Therefore, IR spectroscopy is a useful analytical tool for determining the molecular structure of molecules.
Different compounds display unique properties due to their...
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IR Frequency Region: Fingerprint Region01:03

IR Frequency Region: Fingerprint Region

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IR spectra are divided into two main regions: the diagnostic region and the fingerprint region. The diagnostic region of the spectrum lies above 1500 cm−1. The absorptions resulting from single-bond vibrations of the N–H, C–H, and O–H stretch at higher wavenumbers and appear on the left side of the spectrum. The stretching absorptions of the C≡C and C≡N occur between 2100–2300 cm−1. In contrast, those arising from stretching absorptions of the...
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Raman Spectroscopy: Overview01:20

Raman Spectroscopy: Overview

804
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

Super-resolution Fluorescence Microscopy

11.0K
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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Related Experiment Video

Updated: Oct 26, 2025

High-definition Fourier Transform Infrared FT-IR Spectroscopic Imaging of Human Tissue Sections towards Improving Pathology
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Multiplexed Fourier Transform Infrared and Raman Imaging.

Guillermo Quintás1, Bayden R Wood2, Hugh J Byrne3

  • 1Health & Biomedicine, LEITAT Technological Center, Barcelona, Spain.

Methods in Molecular Biology (Clifton, N.J.)
|July 31, 2021
PubMed
Summary

Multiplexed infrared (IR) and Raman imaging offer comprehensive label-free analysis of biochemical samples. This summary outlines methods to overcome technical challenges and extract vital chemical information from combined vibrational spectra.

Keywords:
Data fusionInfraredLabel-free imagingMultimodal imagingRaman

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Area of Science:

  • Biophysical techniques
  • Vibrational spectroscopy
  • Chemical imaging

Background:

  • Infrared (IR) and Raman spectroscopies are label-free vibrational imaging techniques.
  • These methods provide complementary chemical information about biological samples.
  • Integrating IR and Raman imaging enhances sample analysis.

Purpose of the Study:

  • To summarize the steps for performing multiplexed IR and Raman imaging.
  • To identify and address challenges in combining these techniques.
  • To facilitate comprehensive analysis of biochemical samples.

Main Methods:

  • Detailed procedural steps for multiplexed IR and Raman imaging.
  • Strategies for overcoming technical incompatibilities between IR and Raman.
  • Methods for extracting information from large spectral datasets.

Main Results:

  • Successful integration of IR and Raman imaging is demonstrated.
  • Solutions are presented for technical challenges in multiplexing.
  • Effective data analysis strategies for combined spectra are outlined.

Conclusions:

  • Multiplexed IR and Raman imaging provides a powerful, label-free approach.
  • Overcoming technical hurdles enables comprehensive biochemical analysis.
  • Integrated vibrational spectroscopy enhances understanding of sample composition.