Abstract
We present a compressive lens-free technique that performs tomographic imaging across a cubic millimeter-scale volume from highly sparse data. Compared with existing lens-free 3D microscopy systems, our method requires an order of magnitude fewer multi-angle illuminations for tomographic reconstruction, leading to a compact, cost-effective and scanning-free setup with a reduced data acquisition time to enable high-throughput 3D imaging of dynamic biological processes. We apply a fast proximal gradient algorithm with composite regularization to address the ill-posed tomographic inverse problem. Using simulated data, we show that the proposed method can achieve a reconstruction speed ∼10× faster than the state-of-the-art inverse problem approach in 3D lens-free microscopy. We experimentally validate the effectiveness of our method by imaging a resolution test chart and polystyrene beads, demonstrating its capability to resolve micron-size features in both lateral and axial directions. Furthermore, tomographic reconstruction results of neuronspheres and intestinal organoids reveal the potential of this 3D imaging technique for high-resolution and high-throughput biological applications.
Highlights
Imaging and analysis of 3-dimensional (3D) cell cultures has led to new opportunities in stem cell research, personalized medicine for cancer or hereditary disorders and drug discovery [1,2,3,4]
The region of interest is discretized with a 100 × 100 × 100 grid whose pixel size is 80 nm
Plane waves with a wavelength of 600 nm in vacuum are utilized to illuminate the phantom from 4 directions as described in the experimental setup section
Summary
Imaging and analysis of 3-dimensional (3D) cell cultures has led to new opportunities in stem cell research, personalized medicine for cancer or hereditary disorders and drug discovery [1,2,3,4]. Confocal fluorescence microscopy is currently one of the main tools for 3D cell imaging [5,6]. It requires pointwise scanning of the sample with a focused laser beam, leading to photobleaching and phototoxicity which prevents this technique from being used in long term live cell imaging [7]. Through exploiting the RI contrast of the intrinsic cellular or tissue structures, ODT has shown its potential in cell pathophysiology [17,18,19,20,21].
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