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Related Concept Videos

IR Spectroscopy: Molecular Vibration Overview01:24

IR Spectroscopy: Molecular Vibration Overview

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When Infrared (IR) radiation passes through a covalently bonded molecule, the bonds transition from lower to higher vibrational levels. The fundamental vibrational motions that result in infrared absorption can be classified as stretching or bending vibrations.
Stretching vibrations are vibrational motions that occur along the bond line, changing the bond length or distance between two bonded atoms. They are further distinguished as symmetric or asymmetric. In symmetric stretching, the...
5.3K
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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UV–Vis Spectroscopy: Molecular Electronic Transitions01:16

UV–Vis Spectroscopy: Molecular Electronic Transitions

3.2K
In Ultraviolet–Visible (UV–Vis) spectroscopy, the absorption of electromagnetic radiation is used to probe the electronic structure of molecules. This technique provides insights into molecular electronic transitions, particularly the movement of electrons between different molecular orbitals. Radiation is absorbed if the energy of the electromagnetic radiation passing through the molecule is precisely equal to the energy difference between the excited and ground states. During this...
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Tandem Mass Spectrometry01:21

Tandem Mass Spectrometry

2.8K
Tandem mass spectrometry is a technique that uses multiple mass analyzers in series to obtain a higher selectivity and reduce chemical noise during analyte detection. Instruments with multiple analyzers separated by an interaction cell enable secondary fragmentation and selected study of the fragment ions.Secondary fragmentations occur in the interaction cell and can be induced by various factors. Fragmentation induced by collision with inert gases, such as N2, Ar, He, etc., is called...
2.8K
IR Spectrum Peak Splitting: Symmetric vs Asymmetric Vibrations01:08

IR Spectrum Peak Splitting: Symmetric vs Asymmetric Vibrations

2.0K
Identical bonds within a polyatomic group can stretch symmetrically (in-phase) or asymmetrically (out-of-phase). Similar to hydrogen bonding, these vibrations also influence the shape of the IR peak. Generally, asymmetric stretching frequencies are higher than symmetric stretching frequencies. For example, primary amines exhibit two distinct IR peaks between 3300–3500 cm−1 corresponding to the symmetric and asymmetric N-H stretching, while secondary amines exhibit a single...
2.0K
Raman Spectroscopy: Overview01:20

Raman Spectroscopy: Overview

2.0K
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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Author Spotlight: Unveiling the Potential of VSFG Microscopy in Studying Mesoscopically Heterogeneous Self-Assembled Structures
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Super-multiplex vibrational imaging.

Lu Wei1, Zhixing Chen1, Lixue Shi1

  • 1Department of Chemistry, Columbia University, New York, New York 10027, USA.

Nature
|April 21, 2017
PubMed
Summary
This summary is machine-generated.

Researchers developed a new super-multiplex optical imaging method using stimulated Raman scattering. This technique allows for high-sensitivity, high-selectivity visualization of 24 distinct molecular species in live cells, advancing biological research.

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Area of Science:

  • Biophysics
  • Molecular Imaging
  • Cell Biology

Background:

  • Current molecular imaging techniques face limitations in simultaneously visualizing numerous distinct molecular species within cells due to spectral overlap and sensitivity issues.
  • Fluorescence microscopy is constrained by a 'colour barrier,' limiting the number of distinguishable signals, while spontaneous Raman microscopy suffers from weak signal intensity.
  • Existing methods struggle to achieve high selectivity and sensitivity for imaging multiple molecular targets quantitatively within live biological systems.

Purpose of the Study:

  • To develop a novel optical imaging approach capable of visualizing a large number of distinct molecular species within live cells with high selectivity and sensitivity.
  • To overcome the limitations of existing microscopy techniques, particularly the 'colour barrier' in fluorescence and low signal in spontaneous Raman microscopy.
  • To establish a super-multiplex imaging capability for detailed analysis of cellular and tissue heterogeneity.

Main Methods:

  • Utilized stimulated Raman scattering (SRS) microscopy under electronic pre-resonance conditions for enhanced signal detection.
  • Developed a palette of triple-bond-conjugated near-infrared dyes, each exhibiting a single peak in the cell-silent Raman spectral window.
  • Combined the novel dye palette with existing fluorescent probes to achieve 24 resolvable colours for super-multiplex imaging.

Main Results:

  • Achieved highly selective and sensitive imaging of target molecules in live cells, with sensitivity down to 250 nanomolar and a time constant of 1 millisecond.
  • Demonstrated a 24-colour super-multiplex imaging capability by combining novel Raman dyes with fluorescent probes.
  • Successfully visualized cell-type-dependent heterogeneities in DNA and protein metabolism in neuronal co-cultures and brain tissues.

Conclusions:

  • The developed stimulated Raman scattering-based super-multiplex optical imaging approach offers unprecedented capability for visualizing multiple molecular species in live biological systems.
  • This technique overcomes previous spectral and sensitivity limitations, enabling deeper insights into complex cellular processes and tissue heterogeneity.
  • The 24-colour imaging platform holds significant potential for advancing our understanding of intricate interactions in physiology and pathology.