A correlative quantitative phase contrast and fluorescence super-resolution microscope for imaging molecules in their cellular context
Bao, Y.; Marin, Z.; Chen, X.; Liu, Q.; Tuna, Y.; Stoller, S.; Schueder, F.; Zhang, C.; Fung, S. Y. S.; Wu, M.; M. Neugebauer, K.; Howard, J.; Liu, C.; Baddeley, D.; Shribak, M.; Bewersdorf, J.
Show abstract
Fluorescence microscopy has been widely used to reveal the spatial distribution of specifically labeled molecules, but it is blind to cellular context. Quantitative phase contrast microscopy (QPC) provides such complementary information. Here we have developed a platform that combines the QPC technique of correlative orientation-independent differential interference contrast (OI-DIC) microscopy with single-molecule super-resolution fluorescence microscopy. We demonstrate a detection sensitivity of 0.05 nm optical path difference, sufficient to detect single microtubules, and show its capability of 3D super-resolution fluorescence imaging in the cellular context. Additionally, we report deep-learning enabled digital staining, identifying nuclei, mitochondria and lipid droplets from OI-DIC data and demonstrate the potential of this approach for long-term live-cell imaging of organelles of interest without the need for fluorescence. OI-DIC can be easily integrated into most fluorescence microscopes and is readily adoptable by microscopy labs. ONE-SENTENCE TEASERA highly sensitive technique to visualize sub-cellular structures, dynamics, and molecules in their cellular context.
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