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Ratiometric imaging of minor groove binders in mammalian cells using Raman microscopy.
Christian Tentellino1, William J Tipping1, Leah M C McGee2
1Centre for Molecular Nanometrology, WestCHEM, Department of Pure and Applied Chemistry, Technology and Innovation Centre, University of Strathclyde Glasgow G1 1RD UK karen.faulds@strath.ac.uk duncan.graham@strath.ac.uk.
RSC Chemical Biology
|December 22, 2022
Summary
Raman microscopy enables semi-quantitative imaging of novel DNA minor groove binders (S-MGBs) in live cells. S-MGB-528 showed greater intracellular accumulation than S-MGB-529, highlighting imaging
Area of Science:
- * Pharmacology
- * Chemical Biology
- * Biophysics
Background:
- * Quantitative drug imaging in live cells is crucial for drug discovery but challenging.
- * Solution-based screening methods overlook cellular environment impacts on drug-cell interactions.
- * Raman microscopy offers a powerful tool for visualizing drug behavior within cells.
Purpose of the Study:
- * To semi-quantitatively image novel Strathclyde-minor groove binders (S-MGBs) in mammalian cells using Raman microscopy.
- * To investigate the influence of tail group chemistry (pKa) on S-MGB intracellular uptake, distribution, and mechanism.
- * To compare the cellular behavior of S-MGB-528 and S-MGB-529.
Main Methods:
- * Raman microscopy and stimulated Raman scattering (SRS) microscopy for semi-quantitative drug imaging.
- * Synthesis of novel S-MGBs with alkyne moieties for analysis in the cell-silent Raman region.
- * Multimodal imaging to track intracellular localization and accumulation.
Main Results:
- * Raman imaging revealed differential intracellular accumulation of S-MGB-528 and S-MGB-529.
- * S-MGB-528 demonstrated greater intracellular accumulation despite lower DNA binding affinity in solution compared to S-MGB-529.
- * SRS microscopy showed S-MGB-528 initially localized in the nucleus before lysosomal accumulation.
Conclusions:
- * Raman spectroscopy is a valuable technique for semi-quantitative drug imaging in live cells.
- * Cellular environment significantly impacts drug behavior, influencing accumulation and localization.
- * Imaging techniques are essential for understanding drug-cell interactions and optimizing drug design.
