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We use photothermal induced resonance (PTIR) imaging and spectroscopy, inresonant and non-resonant mode, to study the cytoplasmic membrane and surfaceof intact cells. Non-resonant PTIR images apparently provide rich details ofthe cell surface. However we show that non-resonant image contrast does notarise from the infrared absorption of surface molecules and is insteaddominated by the mechanics of tip-sample contact. In contrast, spectra andimages of the cellular surface can be selectively obtained by tuning thepulsing structure of the laser to restrict thermal wave penetration to thesurface layer. Resonant PTIR images reveal surface structures and domains thatrange in size from about 20 nm to 1 um and are associated to the cytoplasmicmembrane and its proximity. Resonant PTIR spectra of the cell surface arecomparable to far-field IR spectra and provide the first selective measurementof the IR absorption spectrum of the cellular membrane of an intact cell. Inresonant PTIR images, signal intensity, and therefore contrast, can be ascribedto a variety of factors, including mechanical, thermodynamic and spectroscopicproperties of the cellular surface. While PTIR images are difficult tointerpret in terms of spectroscopic absorption, they are easy to collect andprovide unique contrast mechanisms without any exogenous labeling. As such theyprovide a new paradigm in cellular imaging and membrane biology and can be usedto address a range of critical questions, from the nature of membrane lipiddomains to viral membrane fusion.

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