Infrared Scattering-Type Scanning Near-Field Optical Microscopy in Water

Infrared (IR) absorption spectroscopy detects state and chemical composition of biomolecules solely by their inherent vibrational fingerprints. Major disadvantages like the lack of spatial resolution and sensitivity were compensated lately by the use of pointed probes as local sensors enabling the detection of quantities as few as hundreds of proteins with nanometer precision. This makes infrared scattering-type scanning near-field optical microscopy a very powerful tool in life science. The strong absorption of infrared radiation of liquid water, however, still prevents to simply access the measured quantity – light scattered at the probing atomic force microscope tip. Here we report on the local IR response of biological membranes immersed in aqueous bulk solution. We make use of a silicon solid immersion lens as substrate and focusing optics to achieve detection efficiencies sufficient to yield IR near-field maps of purple membranes. We scrutinized our experimental findings by applying theoretical models. Finally, we suggest a means to improve the imaging quality by laser scanning assisted scattering-type scanning near-field optical microscopy. We believe that IR scattering-type scanning near-field optical microscopy will resolve biological structures in their native environments at nm resolution without the need for labeling.