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Background-Suppressed High-Throughput Mid-Infrared Photothermal Microscopy via Pupil Engineering

2021/04/13 by Zong, Haonan, Yurdakul, Celalettin, Bai, Yeran +3 · 1 citation
#Applied Physics (physics.app-ph) #FOS: Physical sciences #Optics (physics.optics)

paper · doi:10.48550/arxiv.2104.06247

Abstract

Mid-infrared photothermal (MIP) microscopy has been a promising label-free chemical imaging technique for functional characterization of specimens owing to its enhanced spatial resolution and high specificity. Recently developed wide-field MIP imaging modalities have drastically improved speed and enabled high-throughput imaging of micron-scale subjects. However, the weakly scattered signal from sub-wavelength particles becomes indistinguishable from the shot-noise as a consequence of the strong background light, leading to limited sensitivity. Here, we demonstrate background-suppressed chemical fingerprinting at a single nanoparticle level by selectively attenuating the reflected light through pupil engineering in the collection path. Our technique provides over three orders of magnitude background suppression by quasi-darkfield illumination in epi-configuration without sacrificing lateral resolution. We demonstrate 6-fold signal-to-background noise ratio improvement, allowing for simultaneous detection and discrimination of hundreds of nanoparticles across a field of view of 70 um x 70 um. A comprehensive theoretical framework for photothermal image formation is provided and experimentally validated with 300 and 500~nm PMMA beads. The versatility and utility of our technique are demonstrated via hyperspectral dark-field MIP imaging of S. aureus and E. coli bacteria.

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