Mid-Infrared Spectroscopy and Microscopy of Subcellular Structures in Eukaryotic Cells with Atomic Force Microscopy - Infrared Spectroscopy

TL;DR

This study demonstrates the use of AFM-IR spectroscopy for high-resolution, nanoscale imaging and spectral analysis of subcellular structures in fixed eukaryotic cells, revealing molecular composition and new nonlinear features.

Contribution

It introduces AFM-IR as a tool for subcellular spectroscopy with spatial resolution below 50 nm, enabling detailed molecular analysis at the nanoscale.

Findings

AFM-IR images of lipid droplets, vesicles, and cytoskeletal filaments were obtained.

Spectra showed characteristic IR absorption bands of phospholipids and polypeptides.

Nonlinear photothermal spectral features were observed, suggesting enhanced sensitivity.

Abstract

Atomic Force Microscopy - Infrared (AFM-IR) spectroscopy allows spectroscopic studies in the mid-infrared spectral region with a spatial resolution better than 50 nm. We show that the high spatial resolution can be used to perform spectroscopic and imaging studies at the subcellular level in fixed eukaryotic cells. We collect AFM-IR images of subcellular structures that include lipid droplets, vesicles and cytoskeletal filaments, by relying on the intrinsic contrast from IR light absorption. We also obtain AFM-IR absorption spectra of individual structures. Most spectra show features that are recognizable in the IR absorption spectra of cells and tissue obtained with FTIR technology, including absorption bands characteristic of phospholipids and polypeptides. We also observe sharp spectral features that we attribute to the nonlinear photothermal response of the system and we propose that they can be used to perform sensitive spectroscopy on the nanoscale.