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Spatiotemporal-multiplexed Fourier ptychographic diffraction tomography for high-speed, label-free 3D imaging of live cells
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DOI：10.1117/1.AP.8.2.026003.png)
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Abstract
En 中文
Label-free, three-dimensional (3D) imaging of organelle dynamics at high spatiotemporal resolution is essential for deciphering cellular physiology. While optical diffraction tomography (ODT) can retrieve intrinsic-contrast 3D refractive-index distribution of transparent cells, its sequential, variable-angle acquisition compromises temporal resolution, obscuring rapid events such as membrane blebbing, vesicle trafficking, and cytoskeletal remodeling. Here, we introduce spatiotemporal-multiplexed Fourier ptychographic diffraction tomography (STM-FPDT), a noninterferometric ODT modality that employs hybrid coherent/partially coherent annular illumination to achieve high resolution in both space (x-y-z) and time (t). By integrating a spatiotemporal reconstruction framework with a sliding window protocol and nonlinear global optimization, STM-FPDT achieves volumetric imaging at 5 Hz with 347 nm lateral and 1.54 mu m axial resolution over a 40 & times;/0.75 NA objective. We validate STM-FPDT across diverse cell lines, successfully capturing membrane blebbing, mitochondrial fission, and intercellular interactions that evade conventional ODT. Readily retrofittable to standard bright-field microscopes, STM-FPDT offers a broadly accessible platform for quantitative, high-spatiotemporal-resolution studies of live-cell physiology.
Keywords:
diffraction tomography
live-cell imaging
refractive index imaging
3D microscopy
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