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Related Concept Videos

Applications of IR Spectroscopy: Overview01:11

Applications of IR Spectroscopy: Overview

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The non-destructive nature and ability to provide valuable chemical information make IR spectroscopy a versatile technique with broad applications in various scientific and industrial fields. IR spectroscopy is commonly used to identify and characterize organic and inorganic compounds. It provides information about the functional groups present in a molecule and the bonding between atoms. This helps in the structural elucidation of compounds during organic synthesis, pharmaceutical research,...
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UV–Vis Spectrometers01:14

UV–Vis Spectrometers

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The absorbance of UV and visible (UV–visible) radiations is measured using a UV–visible spectrophotometer. Deuterium lamps, which emit UV radiation, and tungsten lamps, which produce radiation in the visible region, are used as light sources in UV–visible spectrophotometers. A monochromator or prism is used for diffraction grating, i.e., to split the incoming radiation into different wavelengths. A system of slits is used to focus the desired wavelength on the sample cell.
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UV–Vis Spectroscopy: Molecular Electronic Transitions01:16

UV–Vis Spectroscopy: Molecular Electronic Transitions

2.1K
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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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...
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Ultraviolet and Visible (UV–Vis) Spectroscopy: Overview01:02

Ultraviolet and Visible (UV–Vis) Spectroscopy: Overview

3.7K
Ultraviolet–visible (UV–visible or UV–Vis) spectroscopy is an analytical technique that investigates the interaction between matter and UV–Vis light within the electromagnetic spectrum. This method is widely used for its versatility, simplicity, and relatively quick data acquisition, making it valuable for both qualitative and quantitative analysis. When UV–Vis radiation passes through a material,  molecules absorb light depending on the energy required for...
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Infrared (IR) Spectroscopy: Overview01:09

Infrared (IR) Spectroscopy: Overview

3.4K
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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Related Experiment Video

Updated: Oct 29, 2025

In vivo Imaging of Biological Tissues with Combined Two-Photon Fluorescence and Stimulated Raman Scattering Microscopy
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In vivo Imaging of Biological Tissues with Combined Two-Photon Fluorescence and Stimulated Raman Scattering Microscopy

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In Vivo Spectroscopy.

Cornelia Klose1

  • 1Faculty of Biology, Institute of Biology II, University of Freiburg, Freiburg, Germany. cornelia.klose@biologie.uni-freiburg.de.

Methods in Molecular Biology (Clifton, N.J.)
|July 19, 2019
PubMed
Summary

This study details in vivo spectroscopy for assaying plant phytochromes. The method uses light-induced absorbance changes to quantify total photoreversible phytochromes and the Pfr conformation in intact plant tissues.

Area of Science:

  • Plant physiology
  • Spectroscopy
  • Biophysics

Background:

  • Phytochromes are crucial photoreceptors regulating plant growth and development.
  • Directly assaying phytochromes in intact plant material presents challenges due to light scattering.
  • Existing methods often require tissue extraction, which can alter phytochrome conformation.

Purpose of the Study:

  • To describe an in vivo spectroscopy method for directly assaying phytochromes in intact plants.
  • To detail the instrument setup and measurement procedure for automated dual-wavelength ratio spectrophotometers (ratiospects).
  • To explain data calculation for determining total photoreversible phytochromes and the Pfr phytochrome proportion.

Main Methods:

  • Utilizing in vivo spectroscopy on intact plant material.
Keywords:
In vivo spectroscopyPhotoconversionPhotoreversibilityPhytochromeRatiospectRatiospectrophotometer

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  • Employing dual-wavelength ratio spectrophotometers (ratiospects) to overcome light scattering issues.
  • Applying actinic irradiation to induce photoreversible absorbance changes in phytochromes.

    • Main Results:

    • Demonstration of a reliable method for in vivo phytochrome assay in scattering plant tissues.
    • Quantification of total photoreversible phytochromes is achievable.
    • Determination of the proportion of phytochrome in the biologically active Pfr conformation is possible.

    Conclusions:

    • In vivo spectroscopy with ratiospects provides a non-destructive method for phytochrome analysis.
    • The described method allows for accurate assessment of phytochrome status directly within the plant.
    • This technique is valuable for understanding plant photomorphogenesis and responses to light.