We demonstrate a high-speed confocal-based interferometric scattering (iSCAT) microscopy for label-free live cell imaging. Using a spinning disk scanning unit, our wide-field iSCAT confocal microscope reaches a high image acquisition rate of 1,000 frames per second. Nano-sized objects, as small as 10 nm gold nanoparticles, are directly visualized by the iSCAT confocal microscope in scattering. With the high spatiotemporal resolutions and the high sensitivity, we explore the complex organization and dynamics in live cells.
We present a novel optical microscope technique to detect the chromatin organization and DNA damage foci in the unlabeled live cell nuclei. The dynamic scattering signal of chromatin is recorded by a highly sensitive interference microscopy, contrast-enhanced coherent brightfield microscopy (COBRI), at a high speed. By analyzing the temporal fluctuations of the scattering signal, the density and the compaction level of chromatin are accurately estimated with the sub-micrometer spatial resolutions. This imaging strategy is referred to as “DYNAMICS imaging”. Using DYNAMICS imaging, we monitor the dynamic chromatin remodeling initiated by DNA damage.
Current optical microscope imaging heavily relies on fluorescence-based detection because of its high detection
sensitivity and molecular specificity through labeling. However, the photophysics and photochemistry of the fluorophores,
e.g., photobleaching, blinking and saturation, have limited its applicability to long-term observation and quantitative
imaging. Label-free interference microscopy potentially overcomes the challenges aforementioned by measuring the
scattered field of the sample quantitatively, but these approaches usually compromise on the sensitivity and molecular
specificity of imaging. Here, we demonstrate coherent brightfield (COBRI) microscopy, a highly sensitive scatteringbased
interference microscopy, to capture the dynamic linear scattering signal of live cell nucleus. We demonstrate a
strategy to reconstruct the nuclear organization from the time-lapse COBRI imaging. Our method offers the opportunity
to investigate the cell nuclear dynamics at the unprecedented spatial and temporal resolutions.
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