This study presents a novel label-free approach for characterizing cell death states, eliminating the need for complex molecular labeling that may yield artificial or ambiguous results due to technical limitations in microscope resolution. The proposed holographic tomography technique offers a label-free avenue for capturing precise three-dimensional (3D) refractive index morphologies of cells and directly analyzing cellular parameters like area, height, volume, and nucleus/cytoplasm ratio within the 3D cellular model. We showcase holographic tomography results illustrating various cell death types and elucidate distinctive refractive index correlations with specific cell morphologies complemented by biochemical assays to verify cell death states.
View Article and Find Full Text PDFData acquisition and processing is a critical issue for high-speed applications, especially in three-dimensional live cell imaging and analysis. This paper focuses on sparse-data sample rotation tomographic reconstruction and analysis with several noise-reduction techniques. For the sample rotation experiments, a live Candida rugosa sample is used and controlled by holographic optical tweezers, and the transmitted complex wavefronts of the sample are recorded with digital holographic microscopy.
View Article and Find Full Text PDFHolographic tomography (HT) is an advanced label-free optical microscopic imaging method used for biological studies. HT uses digital holographic microscopy to record the complex amplitudes of a biological sample as digital holograms and then numerically reconstruct the sample's refractive index (RI) distribution in three dimensions. The RI values are a key parameter for label-free bio-examination, which correlate with metabolic activities and spatiotemporal distribution of biophysical parameters of cells and their internal organelles, tissues, and small-scale biological objects.
View Article and Find Full Text PDFIn this study, a novel adaptive wavefront correction (AWC) technique is implemented on a compactly developed structured illumination holographic tomography (SI-HT) system. We propose a mechanical movement-free compact scanning architecture for SI-HT systems with AWC, implemented by designing and displaying a series of computer-generated holograms (CGH) composed of blazed grating with phase Fresnel lens on a phase-only spatial light modulator (SLM). In the proposed SI-HT, the aberrations of the optical system are sensed by digital holography and are used to design the CGH-based AWC to compensate the phase aberrations of the tomographic imaging system.
View Article and Find Full Text PDFThis paper proposes one-shot synthetic aperture digital holographic microscopy using a combination of angular-multiplexing and coherence gating. The proposed angular-multiplexing technique uses multiple noncoplanar incident beams into the synthetic aperture to create tight packed passbands so as to extend spatial frequency spectrum. Coherence gating is performed to prevent the self-interference among the multiple beams.
View Article and Find Full Text PDFDigital 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.
View Article and Find Full Text PDFThis 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).
View Article and Find Full Text PDFThis 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.
View Article and Find Full Text PDFThis study describes the overlapping of spatial frequency bands for synthetic aperture in digital holography using spectrum normalization to effectively enhance the spatial resolutions of image reconstruction. Synthesized spectrum swelling induced by excessive frequency overlaps can be normalized through the inverse apodization of an adjustable window function, which is similar to the effects of suppressing low-frequency expansion and strengthening high-frequency components of the reconstructed images. The results indicated that using the normalized spectrum synthesis that requires only a few frequency bands effectively enhances the spatial resolution and phase sensitivity of reconstructed images in digital holographic microscopy.
View Article and Find Full Text PDFBased on scalar diffraction theory and the geometric structure of liquid crystal on silicon (LCoS), we study the impulse responses and image depth of focus in a holographic three-dimensional (3D) display system. Theoretical expressions of the impulse response and the depth of focus of reconstructed 3D images are obtained, and experimental verifications of the imaging properties are performed. The results indicated that the images formed by holographic display based on the LCoS device were periodic image fields surrounding optical axes.
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