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Label-free in vitro visualization and characterization of caveolar bulbs during stimulated re-epithelialization

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Abstract

Tip-enhanced Raman scattering (TERS) was paired with real-time reverse transcription quantitative polymerase chain reaction (RT-qPCR) to characterize lipid aggregates during stimulated re-epithelialization using an in vitro wound healing model. In this study, lipid fluctuations in the plasma membrane of epidermal keratinocytes were studied at multiple time points post-wounding. TERS measurements for the first time were also combined with sample analysis after initial wounding and 24 h of wound healing. This enabled simultaneous visualization and characterization of caveolar bulb distribution during wound healing stages, providing noninvasive insight into their associated lipid structure and coating protein, caveolin, in the nanometer range. The combination of Raman spectroscopy and scanning probe microscopy in TERS gives access to topographic and chemical structure information in a single experiment. It is the intrinsic specificity and sensitivity of TERS that enable this discrete detection of cell surface components on the nanometer scale. In contrast with competing biochemical methods, the applied technique does not interfere with the cellular composition, enabling lipid structure analysis without digestion or detergents, and displayed great potential for future biological in vivo studies.

tip_enhanced Raman spectroscopy of cell caveolar bulbs formed on a wound healing model

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Acknowledgments

We thank the Bundesministerium für Bildung und Forschung (No. 0312032B and No. 0312032C) for their financial support.

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Authors and Affiliations

  1. East 42nd Street, New York, NY, 10017, USA

    Melissa Watkins-Mariani

  2. IPHT–Leibniz Institute for Photonic Technology, Albert-Einstein-Str. 9, 07743, Jena, Germany

    Tanja Deckert-Gaudig & Volker Deckert

  3. IPC–Friedrich-Schiller-University of Jena and Abbe Center of Photonics, Helmholtzstr. 4, 07743, Jena, Germany

    Volker Deckert

Authors
  1. Melissa Watkins-Mariani
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  2. Tanja Deckert-Gaudig
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  3. Volker Deckert
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Correspondence to Volker Deckert.

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Published in the topical collection Single Cell Analysis with guest editors Petra Dittrich and Norbert Jakubowski.

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Watkins-Mariani, M., Deckert-Gaudig, T. & Deckert, V. Label-free in vitro visualization and characterization of caveolar bulbs during stimulated re-epithelialization. Anal Bioanal Chem 406, 6993–7002 (2014). https://doi.org/10.1007/s00216-014-7998-y

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  • DOI: https://doi.org/10.1007/s00216-014-7998-y

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