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Two-photon photopolymerization (TPP) has recently become a popular method for the fabrication of three-dimensional (3D) micro- and nanostructures. The reproduction fidelity of the designed micro- and nanostructures is influenced by experimental writing conditions, including laser power, exposure time, etc. To determine the appropriate writing parameters, characterization of morphological features and surface roughness during the experiment is needed. Traditional characterization methods for TPP, e.g., scanning electron microscopy and atomic force microscopy, have limited speed and cannot study internal structures without invasive approaches. Optical diffraction tomography (ODT) is an emerging label-free 3D imaging technique based on reconstructing the object's 3D refractive index (RI) distribution with diffraction-limited resolution. Here, we propose a non-invasive solution to fully characterize the TPP-fabricated structures using a high-speed ODT technique, which can eliminate the need for complex sample preparation, such as fluorescence labelling or metal-coating, and achieve a full 3D measurement time of 6 ms. By visualizing and studying different TPP-fabricated structures, including embedded spirals and cubes, via the ODT system, the fabrication quality, including 3D morphological features, exposure levels, and surface roughness, can be examined quantitatively. The results suggest our method can effectively improve the fabrication quality and reproducibility of TPP, generating impacts on the nanofabrication community.

Three-dimensional (3D) image cytometers may significantly improve the cell analysis accuracy to facilitate biological discoveries and clinical diagnosis, but their development is curbed by the low imaging throughput. Here we report SIngle-frame LAbel-free Cell Tomography (SILACT) with diffraction-limited resolution and unprecedented imaging speed of over 10,000 volumes/second. SILACT is built on a unique interferometric microscope with angle-multiplexing illumination and a pre-trained physics-incorporating Deep Neural Network for efficient 3D Refractive Index (RI) reconstruction, from which 3D morphological and biophysical parameters of cells are extracted. With microfluidics and a high-speed camera, SILACT is capable of imaging over 20,000 cells/second and distinguishing different cell species during rapid measurements of large cell quantities, as well as visualizing shear-induced 3D transient deformation of red blood cells on a sub-millisecond scale., Comment: Baoliang Ge, Yanping He, Mo Deng contributed equally to this work