Imaging and Spectroscopy of Domains of the Cellular Membrane by Photothermal-Induced Resonance

TL;DR

This paper demonstrates the use of photothermal induced resonance (PTIR) imaging and spectroscopy to visualize and analyze cellular membrane domains with nanoscale resolution, revealing new contrast mechanisms without labels.

Contribution

It introduces a novel application of resonant PTIR for selective imaging and spectroscopy of cellular membranes, providing first-of-its-kind IR spectra of intact cell membranes.

Findings

Resonant PTIR images reveal membrane domains from 20 nm to 1 μm.

Spectra obtained are comparable to far-field IR spectra.

PTIR provides label-free contrast based on mechanical, thermodynamic, and spectroscopic properties.

Abstract

We use photothermal induced resonance (PTIR) imaging and spectroscopy, in resonant and non-resonant mode, to study the cytoplasmic membrane and surface of intact cells. Non-resonant PTIR images apparently provide rich details of the cell surface. However we show that non-resonant image contrast does not arise from the infrared absorption of surface molecules and is instead dominated by the mechanics of tip-sample contact. In contrast, spectra and images of the cellular surface can be selectively obtained by tuning the pulsing structure of the laser to restrict thermal wave penetration to the surface layer. Resonant PTIR images reveal surface structures and domains that range in size from about 20 nm to 1 um and are associated to the cytoplasmic membrane and its proximity. Resonant PTIR spectra of the cell surface are comparable to far-field IR spectra and provide the first selective measurement of the IR absorption spectrum of the cellular membrane of an intact cell. In resonant PTIR images, signal intensity, and therefore contrast, can be ascribed to a variety of factors, including mechanical, thermodynamic and spectroscopic properties of the cellular surface. While PTIR images are difficult to interpret in terms of spectroscopic absorption, they are easy to collect and provide unique contrast mechanisms without any exogenous labeling. As such they provide a new paradigm in cellular imaging and membrane biology and can be used to address a range of critical questions, from the nature of membrane lipid domains to viral membrane fusion.