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Multimodal Optical Imaging Platform for Studying Cellular Metabolism
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Reaction-Based Ratiometric Sensors for Simultaneous Multi-Bio-Analyte Imaging in Living Cells Using Spontaneous Raman

Sujit K Das1, Heqi Xi2, Itsuki Yamamoto2

  • 1Department of Chemical Sciences, Tata Institute of Fundamental Research, Mumbai, Maharashtra, 400005, India.

Angewandte Chemie (International Ed. in English)
|December 29, 2025
PubMed
Summary
This summary is machine-generated.

Researchers developed novel Raman-responsive ratiometric sensors (ABATaRs) for sensitive bio-analyte imaging. These probes overcome sensitivity limitations, enabling faster, multi-analyte imaging in live cells using spontaneous Raman microscopy.

Keywords:
Activity‐based alkyne tag Raman sensors‐ ABATaRsLive cell Raman imagingMultiplexed imagingRatiometric Raman sensingSpontaneous Raman imaging

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Area of Science:

  • Chemical Biology
  • Spectroscopy
  • Bioimaging

Background:

  • Raman-scattering offers multi-analyte imaging potential due to narrow peak-widths.
  • Alkyne/nitrile tags are crucial for Raman probes but suffer from low Raman-scattering cross-sections, limiting sensitivity.
  • Existing Raman-responsive ratiometric sensors struggle with sensitivity on accessible spontaneous Raman platforms.

Purpose of the Study:

  • To develop highly sensitive Raman-responsive ratiometric sensors for spontaneous Raman imaging.
  • To overcome the low sensitivity challenge associated with alkyne/nitrile Raman probes.
  • To demonstrate the versatility of these sensors for imaging various bio-analytes in live cells.

Main Methods:

  • Leveraged the 'push-pull' electronic effect to design novel activity-based alkyne-tag Raman sensors (ABATaRs).
  • Quantified computed and experimental Raman-scattering activities and relative cross-sections compared to benchmarks.
  • Developed cell-permeable, ratiometric ABATaRs for pH, hydrogen peroxide, and copper (Cu) ions.

Main Results:

  • ABATaRs exhibited significantly higher Raman-scattering activities (12-38 times) and experimental Raman intensity (5-22 times) than benchmark EdU.
  • Achieved high relative Raman scattering cross-sections for alkyne stretching (up to 466 vs. DMSO C-H stretching).
  • Successfully imaged physiological and pathophysiological levels of bio-analytes (pH, H2O2, Cu ions) in live cells at low sensor concentrations (1-5 µM) using spontaneous Raman microscopy.
  • Demonstrated simultaneous multi-analyte Raman imaging of Cu ions and hydrogen peroxide in live cells, enhancing imaging speed.

Conclusions:

  • ABATaRs represent a breakthrough in sensitive Raman-based bio-analyte imaging on accessible spontaneous Raman platforms.
  • The developed sensors offer high sensitivity, ratiometric detection, and versatility for various analytes.
  • ABATaRs significantly advance live-cell multi-analyte imaging capabilities, enabling faster and more sensitive detection.