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

TL;DR

This study demonstrates the use of Photothermal-Induced Resonance Infrared Spectroscopy and Imaging to achieve subcellular resolution in intact eukaryotic cells, revealing detailed molecular information without sample drying or fixing.

Contribution

It is the first to apply PTIR spectroscopy to intact, hydrated eukaryotic cells for subcellular imaging and spectral analysis at nanometer resolution.

Findings

Successful PTIR imaging of nuclei and organelles in live cells

Detection of lipid and protein spectral signatures at subcellular level

Variable PTIR signal strength across different cellular structures

Abstract

Photothermal-Induced Resonance (PTIR) spectroscopy and imaging with infrared light has seen increasing application in 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 allowed by light diffraction, typically below 100 nm. In the present work we test the capability of the technique to perform measurements with subcellular resolution on intact eukaryotic cells, without drying or fixing. We demonstrate the possibility to obtain PTIR images and spectra from the nucleus and multiple organelles with high resolution. 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.