Combined Multi-Plane Tomographic Phase Retrieval and Stochastic Optical Fluctuation Imaging for 4D Cell Microscopy

Abstract

Super-resolution fluorescence microscopy provides unprecedented insight into cellular and subcellular structures. However, going "beyond the diffraction barrier" comes at a price since most far-field super-resolution imaging techniques trade temporal for spatial super-resolution. We propose the combination of a novel label-free white light quantitative phase tomography with fluorescence imaging to provide high-speed imaging and spatial super-resolution. The non-iterative phase reconstruction relies on the acquisition of single images at each z-location and thus enables straightforward 3D phase imaging using a classical microscope. We realized multi-plane imaging using a customized prism for the simultaneous acquisition of 8 planes. This allowed us to not only image live cells in 3D at up to 200 Hz, but also to integrate fluorescence super-resolution optical fluctuation imaging within the same optical instrument. This 4D microscope platform unifies the sensitivity and high temporal resolution of phase tomography with the specificity and high spatial resolution of fluorescence imaging.

References in corpus (3)

• Complementarity of PALM and SOFI for super-resolution live cell imaging of focal adhesions
• Structured illumination multimodal 3D-resolved quantitative phase and fluorescence sub-diffraction microscopy
• Quantifying protein densities on cell membranes using super-resolution optical fluctuation imaging

Cited by in corpus (9)

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• Quantitative phase microscopies: accuracy comparison
• Subwavelength Imaging using a Solid-Immersion Diffractive Optical Processor
• Label-free chip-based evanescent light scattering super-resolution and superior-contrast optical microscopy (cELS)
• Optimal correlation order in super-resolution optical fluctuation microscopy
• Autofocusing technologies for whole slide imaging and automated microscopy
• Metasurfaces for infrared multi-modal microscopy: phase contrast and bright field
• Spatial light interference microscopy (SLIM): principle and applications to biomedicine