Abstract
Super-resolution imaging of volumes as large as whole cells in three-dimensions (3D) is required to reveal unknown features of cellular organization which cannot be resolved by conventional fluorescence microscopy. We propose a new 3D high resolution imaging technique based on the principles of single-molecule localization microscopy (SMLM) and fluorescence incoherent correlation holography (FINCH). FINCH enables hologram acquisition and three-dimensional (3D) imaging of large objects emitting incoherent light. This technique combines FINCH and SMLM to enable single-molecule volumetric imaging over large axial ranges without scanning the sample using a simple and robust setup, hence making it a viable solution for whole cell super-resolution imaging of biological samples. Here, we present the underlying theory and simulations demonstrating the extended depth of field. We image a single 0.2-μm fluorescent microsphere using this approach and discuss the signal-to-noise ratio (SNR) requirements for an experimental implementation.
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