Multi-color live-cell super-resolution volume imaging with multi-angle interference microscopy

Abstract

Imaging and tracking of near-surface three-dimensional volumetric nanoscale dynamic processes of live cells remains a challenging problem. In this paper, we propose a multi-color live-cell near-surface-volume super-resolution microscopy method that combines total internal reflection fluorescence structured illumination microscopy with multi-angle evanescent light illumination. We demonstrate that our approach of multi-angle interference microscopy is perfectly adapted to studying subcellular dynamics of mitochondria and microtubule architectures during cell migration.

Imaging of the immune system – towards a subcellular and molecular understanding

“…(Huff, 2016) and STED (12.5 Hz for 2.9×2.9 μm) (Yang et al, 2014), are more suitable for live-cell imaging. SIM is the most commonly used super-resolution imaging technique for live-cell imaging (Chen et al, 2018;Fiolka et al, 2012;Turcotte et al, 2019). SIM has a good imaging speed, can use any labels used in conventional live-cell imaging, and has tolerable light toxicity, similar to conventional live-cell imaging.…”

Far‐Field Superresolution Imaging via Spatial Frequency Modulation

“…Among the existing optical superresolution methodologies, the SIM method [109,165,[189][190][191] is one of the most commonly implemented optical superresolution techniques for studying the dynamic biological process at high speed, based on its widefield implement. In SIM, the sample is illuminated with spatially structured excitation light to visualize normally inaccessible high-resolution information in the form of moiré fringes.…”

“…By combining this exponential decay leveraging method with TIRF-SIM, the multi-angle interference microscopy (MAIM) can provide a sub-100 nm lateral resolution and 40 nm axial resolution over 600 nm depth range. [189] Chip-Based SIM (cSIM): In conventional SIM and TIRF-SIM, a large FOV and high resolution cannot be simultaneously obtained. Besides, the commercial liquid immersion objective lens has a limited NA of ≈1.7, which restricts the resolution of TIRF-SIM to around 80 nm.…”

Correlative cathodoluminescence electron microscopy bioimaging: towards single protein labelling with ultrastructural context

“…[16][17][18] While conventional FM is routinely employed for dynamic live cell imaging, SR-FM is mostly performed on fixed samples due to high laser power, long image acquisition times and lack of suitable probes. 10,19 The most fundamental limitation of optical nanoscopy is the achievable labelling density with respect to the required resolution. Often the required labelling densities are not achievable in biological samples without altering their function.…”