Tunable SIM: observation at varying spatiotemporal resolutions across the FOV

Abstract

To date, imaging systems have generally been designed to provide an even spatiotemporal resolution across the field of view (FOV). However, this becomes a fundamental limitation when we aim to simultaneously observe varying dynamics at different parts of the FOV. In conventional imaging systems, to capture fast dynamics occurring at only a small portion of the FOV, the entire imaging system's sampling rate must be increased. This is a major problem if different parts of the FOV must rather be imaged at high spatial resolutions beyond the diffraction limit and require a sacrifice in temporal resolution. To answer this unmet challenge, we propose tunable SIM, which enables adaptive modulation of spatiotemporally varying structured illumination across different parts of the FOV. Using tunable SIM, we exploit the varying and designable spatiotemporal resolution to demonstrate simultaneous measurements of subdiffraction-limited changes in the actin fine structure of U87MG-EGFP-CD9 cells and the fast viscous flow inducing these structural changes.

How did correlative atomic force microscopy and super-resolution microscopy evolve in the quest for unravelling enigmas in biology?

“…These patterns can be generated either by a projection of a physical grid pattern, or with electronic devices, such as spatial light modulators, [132][133][134] or digital mirror devices. [135][136][137][138] Multiplication of the sample structure with the illumination pattern generates interference patterns in form of Moiré fringes that contain information about the fine details of the sample structure that could not be observed in diffraction-limited imaging. A series of images is acquired with different pattern positions and orientations, and computational techniques are then applied to remove the illumination structure and retrieve high-resolution information.…”