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Optical microscopy is an indispensable tool that is driving progress in biology and is still the only practical means of obtaining spatial and temporal resolution within living cells and tissues. In this context, staining samples with fluorescent labels provides a highly sensitive and specific method of visualizing biomolecules. However, fluorescence microscopy has various limitations including sample manipulation and staining artifacts, fluorophore photobleaching and associated phototoxicity. Therefore, much effort has been devoted to developing label-free optical microscopy techniques which are non-perturbing, photostable, and in turn offer quantitative capabilities unavailable with fluorescent methods.
Our laboratory has been developing quantitative label-free optical microscopy set-ups featuring innovative excitation/detection schemes, with application ranging from synthetic lipid membranes [1] and nanoparticle materials [2-4] to living cells [5]. Specifically, we have demonstrated quantitative differential interference contrast microscopy [1], extinction microscopy [2], four-wave mixing imaging [3,4], and chemically-specific coherent Raman scattering (CRS) microscopy [5-7], including an interferometric CRS set-up which offers background-free image contrast, shot-noise limited detection, and phase sensitivity, enabling topographic imaging of interfaces [8]. We are also developing a new wide-field interferometric reflectometry method, aimed at monitoring single protein-lipid membrane interactions with unprecedented sensitivity. I will present our latest progress with these techniques and their applications to bioimaging.
[1] Anal. Chem. 92, 14657 (2020).
[2] Nanoscale 12, 16215 (2020).
[3] Phys. Rev. X 7, 41022 (2017).
[4] Nanoscale 12, 4622 (2020).
[5] Analyst 146, 2277 (2021).
[6] Nat. Nanotechnol. 9, 940 (2014).
[7] Anal. Chem. 91, 2813 (2019).
[8] APL Photonics 3, 092402 (2018).
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