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Visualizing Bioactive Small Molecules by Alkyne Tagging and Slit-Scanning Raman Microscopy

  • Jun Ando
  • Kosuke Dodo
  • Katsumasa Fujita
  • Mikiko Sodeoka
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1888)

Abstract

An understanding of the intracellular distribution of bioactive small molecules provides insight into their target organelles and biomolecules, and throws light on their molecular mechanisms of action and specificity. Many studies in this area have employed fluorescence imaging, using molecules of interest labeled with fluorescent dyes. However, modification with a bulky fluorophore may significantly alter the properties, including bioactivity, of small molecules. To address this issue, we have recently developed Raman scattering microscopy of alkyne tags to visualize small molecules in biological systems. The tiny alkyne tag should have a minimal effect on the properties of the tagged molecule, and the unique alkyne vibrational properties allow observation of the Raman signals of the tagged molecules in a wavelength region where there is little interference from Raman signals of endogenous biomolecules. Here we summarize the methodology for Raman imaging of alkyne-tagged bioactive small molecules in living biological systems including the development of slit-scanning Raman microscope, which is useful for fast imaging of alkyne-tagged molecules.

Key words

Alkyne tag Raman scattering Small molecules Slit-scanning Raman microscopy 

Notes

Acknowledgment

This work was partially supported by JST-ERATO (Sodeoka Live Cell Chemistry Project), AMED-CREST (No. JP17gm0710004), RIKEN and JSPS KAKENHI Grant Number 26600117 (J.A.).

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Jun Ando
    • 1
  • Kosuke Dodo
    • 2
  • Katsumasa Fujita
    • 1
  • Mikiko Sodeoka
    • 2
  1. 1.Department of Applied PhysicsOsaka UniversitySuitaJapan
  2. 2.Synthetic Organic Chemistry LaboratoryRIKEN Cluster for Pioneering ResearchWakoJapan

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