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Open AccessArticle

Characterization of Intact Eukaryotic Cells with Subcellular Spatial Resolution by Photothermal-Induced Resonance Infrared Spectroscopy and Imaging

by Luca Quaroni 1,2
1
Faculty of Chemistry, Jagiellonian University, ul. Gronostajowa 2, 30-387 Kraków, Poland
2
Institute of Nuclear Physics, Polish Academy of Sciences, ul. Radzikowskiego 152, 31-342 Kraków, Poland
Academic Editor: Kamilla Malek
Molecules 2019, 24(24), 4504; https://doi.org/10.3390/molecules24244504
Received: 29 November 2019 / Accepted: 3 December 2019 / Published: 9 December 2019
Photothermal-induced resonance (PTIR) spectroscopy and imaging with infrared light has seen increasing application in the molecular spectroscopy of biological samples. The appeal of the technique lies in its capability to provide information about IR light absorption at a spatial resolution better than that allowed by light diffraction, typically below 100 nm. In the present work, we tested the capability of the technique to perform measurements with subcellular resolution on intact eukaryotic cells, without drying or fixing. We demonstrate the possibility of obtaining PTIR images and spectra from the nucleus and multiple organelles with high resolution, better than that allowed by diffraction with infrared light. We obtain particularly strong signal from bands typically assigned to acyl lipids and proteins. We also show that while a stronger signal is obtained from some subcellular structures, other large subcellular components provide a weaker or undetectable PTIR response. The mechanism that underlies such variability in response is presently unclear. We propose and discuss different possibilities, addressing thermomechanical, geometrical, and electrical properties of the sample and the presence of cellular water, from which the difference in response may arise. View Full-Text
Keywords: photothermal-induced resonance; PTIR; AFM-IR; infrared spectroscopy; single cell; subcellular imaging; infrared imaging; scanning probe microscopy photothermal-induced resonance; PTIR; AFM-IR; infrared spectroscopy; single cell; subcellular imaging; infrared imaging; scanning probe microscopy
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Quaroni, L. Characterization of Intact Eukaryotic Cells with Subcellular Spatial Resolution by Photothermal-Induced Resonance Infrared Spectroscopy and Imaging. Molecules 2019, 24, 4504.

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