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Updated: Mar 1, 2026
Non-contact, Label-free Monitoring of Cells and Extracellular Matrix using Raman Spectroscopy
Published on: May 29, 2012
Surface-enhanced hyper Raman hyperspectral imaging and probing in animal cells
Zsuzsanna Heiner1, Marina Gühlke2, Vesna Živanović1
1SALSA School of Analytical Sciences Adlershof, Humboldt-Universität zu Berlin, Albert-Einstein-Str. 5-9, 12489 Berlin, Germany and Department of Chemistry, Humboldt-Universität zu Berlin, Brook-Taylor-Str. 2, 12489 Berlin, Germany. janina.kneipp@chemie.hu-berlin.de.
Surface-enhanced hyper Raman scattering (SEHRS) enables detailed imaging of macrophage cells. This advanced technique uses SEHRS labels to reveal subcellular structures and physiological parameters like endosomal pH.
Area of Science:
- Biophotonics
- Spectroscopy
- Cellular Imaging
Background:
- Surface-enhanced hyper Raman scattering (SEHRS) amplifies weak Raman signals from organic molecules near plasmonic nanostructures.
- SEHRS offers advantages over SERS, including complementary vibrational data and probing of small volumes with long-wavelength excitation.
Purpose of the Study:
- To demonstrate SEHRS imaging of macrophage cells using novel SEHRS labels.
- To investigate the potential of SEHRS for mapping subcellular localization and physiological parameters within cells.
Main Methods:
- Utilized silver nanoparticles functionalized with 2-naphthalenethiol and para-mercaptobenzoic acid as SEHRS labels.
- Employed hyperspectral analysis, principal component analysis (PCA), and hierarchical cluster analysis (HCA) for data processing and localization.
- Excited SEHRS labels off-resonance with long wavelengths.
Main Results:
- Successfully imaged macrophage cells using SEHRS, differentiating vibrational signatures of reporter molecules.
- Localized SEHRS probes within subcellular compartments, providing insights into their distribution.
- Demonstrated the potential for mapping endosomal pH variations within the cell cytosol based on SEHRS spectral sensitivity.
Conclusions:
- SEHRS is a powerful technique for high-resolution cellular imaging and probing intracellular environments.
- Hyperspectral SEHRS analysis, combined with PCA and HCA, effectively localizes probes and reveals cellular heterogeneity.
- The sensitivity of SEHRS to environmental changes opens avenues for mapping physiological parameters in live cells.
