This study presents a novel tomographic imaging technique for living biomedical samples using an optically driven full-angle rotation scheme based on digital holographic microscopy, in which the three-dimensional refractive index distribution inside the sample can be measured and analyzed. To accomplish the full-angle sample rotation, two optical traps are driven by highly focused spots on the top and bottom of the sample. The rim image of the sample outside the focal depth at the different rotation angles and propagation distances can be corrected and compensated, respectively, via numerical focusing; therefore, tomographic imaging of the sample can be conducted. The proposed approach shows that an entire symmetric spectrum can be acquired for tomographic reconstruction without the missing apple core problem as in traditional sample-rotation schemes. The three-dimensional refractive index of living yeast in a fluid medium is measured and verified.

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This paper proposes a coded aperture structured illumination (CASI) technique in digital holographic microscopy (DHM). A CASI wave is generated using two binary phase codes (0° and 120°) for spatial phase shifting. The generated CASI wave then interferes with a reference wave to form a coded Fresnel hologram at a single exposure with compressive sensing (CS) to avoid the temporal phase-shifting process of the structured illumination (SI). The CS algorithm is applied to retrieve the missing data of decoded phase-shifted SI-modulated waves, which are used to separate overlapped spatial frequencies for obtaining a larger spatial frequency coverage to provide superresolution imaging. Two phase-only spatial light modulators are applied to generate a directional SI pattern for obtaining a coded aperture with a suitable size to perform one-shot acquisition in the DHM system.

Particle image velocimetry (PIV) is essential in experimental fluid dynamics. In the current work, we propose a new velocity field estimation paradigm, which is a synergetic combination of cross correlation and fully convolutional network (CC-FCN). Specifically, the fully convolutional network is used to optimize and correct a coarse velocity guess to achieve a super-resolution calculation. And the traditional cross correlation method provides the initial velocity field based on a coarse correlation with a large interrogation window. As a reference, the coarse velocity guess helps with improving the robustness of the proposed algorithm. CC-FCN has two types of input layers, one is for the particle images, and the other is for the initial velocity field calculated using cross correlation with a coarse resolution. First, two pyramidal modules extract features of particle images and initial velocity field, respectively. Then the fusion module appropriately fuses these features. Finally, CC-FCN achieves the super-resolution calculation through a series of deconvolution layers to obtain the single-pixel velocity field. As the supervised learning strategy is considered, synthetic data sets including ground-truth fluid motions are generated to train the network parameters. Synthetic and real experimental PIV data sets are used to test the trained neural network in terms of accuracy, precision, spatial resolution and robustness. The test results show that these attributes of CC-FCN are further improved compared with those of other tested PIV algorithms. The proposed model could therefore provide competitive and robust estimations for PIV experiments.

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Digital holographic microtomography is a promising technique for three-dimensional (3D) measurement of the refractive index (RI) profiles of biological specimens. Measurement of the RI distribution of a free-floating single living cell with an isotropic superresolution had not previously been accomplished. To the best of our knowledge, this is the first study focusing on the development of an integrated dual-tomographic (IDT) imaging system for RI measurement of an unlabelled free-floating single living cell with an isotropic superresolution by combining the spatial frequencies of full-angle specimen rotation with those of beam rotation. A novel ‘UFO’ (unidentified flying object) like shaped coherent transfer function is obtained. The IDT imaging system does not require any complex image-processing algorithm for 3D reconstruction. The working principle was successfully demonstrated and a 3D RI profile of a single living cell, Candida rugosa, was obtained with an isotropic superresolution. This technology is expected to set a benchmark for free-floating single live sample measurements without labeling or any special sample preparations for the experiments.

This work describes the influence of coherent illumination on phase measurement accuracy in digital holographic microscopy (DHM). To improve net phase accuracy in a DHM system, the phase referencing and temporal averaging techniques are applied simultaneously to suppress the phase noises caused by the laser source and image sensor. A comparison between a laser diode operated in single- and multi-modes is given to demonstrate the coherence effect on the fringe visibility and the reconstructed phase accuracy of digital holograms. Axial sub-nanometer accuracy in DHM with the fewest successive hologram recordings is performed using a wavelength-stabilized laser diode in single-mode operation.

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