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We propose a multimodal optical system combining ODT and 3-D SIM to measure both 3-D RI and fluorescence distribution of a sample with high spatiotemporal resolution as well as molecular specificity.

Purpose: Scattered radiation from multi‐leaf collimators (MLCs) is no longer negligible for calculating in‐air output ratio, Sc for small and irregular fields often used in intensity‐modulated radiation therapy(IMRT). An extra‐focal source model for scattered radiation from MLCs, namely MLC scatter source, has been developed to improve the accuracy of the Sc calculation. Methods: A conventional dual‐source model was made by using Sc data that were measured for collimator‐defined fields of Varian Clinac IX linear accelerator. Then, an MLC scatter source at the center of the MLC position of the linear accelerator was assumed in the model. The MLC scatter source model consisted of two Gaussian functions of which parameters were iteratively optimized against the Sc data measured for different MLC fields with fixed collimator sizes. To evaluate the effectiveness of the developed source model, measurements were made for various MLC‐defined irregular or square fields. The calculated Sc data by using (1) the developed source model and (2) the conventional dual source model were compared with the measured data. Results: The mean discrepancy between the measured Sc and calculated Sc from the developed source model was 0.08+‐0.28%, while one from the conventional source model was 0.44+−0.39%. Conclusions: The developed MLC scatter source model in conjunction with the dual source model could improve the accuracy of the Sc calculation in IMRT fields.

Microfluidic mixing plays a key role in various fields, including biomedicine and chemical engineering. To date, although various approaches for imaging microfluidic mixing have been proposed, they provide only quantitative imaging capability and require for exogenous labeling agents. Quantitative phase imaging techniques, however, circumvent these problems and offer label-free quantitative information about concentration maps of microfluidic mixing. We present the quantitative phase imaging of microfluidic mixing in various types of PDMS microfluidic channels with different geometries; the feasibility of the present method was validated by comparing it with the results obtained by theoretical calculation based on Fick’s law.

In digital 3D holographic displays, the generation of realistic 3D images has been hindered by limited viewing angle and image size. Here we demonstrate a digital 3D holographic display using volume speckle fields produced by scattering layers in which both the viewing angle and the image size are greatly enhanced. Although volume speckle fields exhibit random distributions, the transmitted speckle fields have a linear and deterministic relationship with the input field. By modulating the incident wavefront with a digital micro-mirror device, volume speckle patterns are controlled to generate 3D images of micrometer-size optical foci with 35° viewing angle in a volume of 2 cm × 2 cm × 2 cm.

Imaging brain tissues is an essential part of neuroscience because understanding brain structure provides relevant information about brain functions and alterations associated with diseases. Magnetic resonance imaging and positron emission tomography exemplify conventional brain imaging tools, but these techniques suffer from low spatial resolution around 100 μm. As a complementary method, histopathology has been utilized with the development of optical microscopy. The traditional method provides the structural information about biological tissues to cellular scales, but relies on labor-intensive staining procedures. With the advances of illumination sources, label-free imaging techniques based on nonlinear interactions, such as multiphoton excitations and Raman scattering, have been applied to molecule-specific histopathology. Nevertheless, these techniques provide limited qualitative information and require a pulsed laser, which is difficult to use for pathologists with no laser training. Here, we present a label-free optical imaging of mouse brain tissues for addressing structural alteration in Alzheimer’s disease. To achieve the mesoscopic, unlabeled tissue images with high contrast and sub-micrometer lateral resolution, we employed holographic microscopy and an automated scanning platform. From the acquired hologram of the brain tissues, we could retrieve scattering coefficients and anisotropies according to the modified scattering-phase theorem. This label-free imaging technique enabled direct access to structural information throughout the tissues with a sub-micrometer lateral resolution and presented a unique means to investigate the structural changes in the optical properties of biological tissues.

Holographic optical tweezers (HOTs) have been utilized for trapping microscopic particles in three dimensions with multiple foci generated by wavefront shaping of light, which can manipulate three-dimensional (3-D) positions of colloidal particles as well as exerting an optical force on particles. So far, most experiments using HOTs have been conducted for trapping spherical particles because optical principles can easily predict optical forces and the responding motion of microspheres. For non-spherical particles, however, calculation of optical forces and torques exerting on samples is very complicated, and the orientation control of non-spherical particles is limited since the non-spherical particles tend to align along the optic axis of the trapping beam. Here, we propose and experimentally demonstrate 3-D trapping of non-spherical particles by wavefront shaping of light based on the measurement of 3-D refractive index (RI) distribution of samples. The 3-D RI distribution of non-spherical particles was measured by optical diffraction tomography and the phase hologram which can generate stable optical traps for the samples was calculated by iterative 3-D Gerchberg-Saxton algorithm from the measured 3-D RI distribution. We first validate the proposed method for stable trapping and orientation control of 2-m colloidal PMMA ellipsoids. The proposed method is also exploited for rotating, folding and assembly of red blood cells.

There is a strong correlation between the dynamic membrane fluctuations and the biomechanical properties of living cells. The dynamic membrane fluctuation consists of submicron displacements, and can be altered by changing the cells’ pathophysiological conditions. These results have significant relevance to the understanding of RBC biophysics and pathology, as follows. RBCs must withstand large mechanical deformations during repeated passages through the microvasculature and the fenestrated walls of the splenic sinusoids. This essential ability is diminished with senescence, resulting in physiological destruction of the aging RBCs. Pathological destruction of the red cells, however, occurs in cells affected by a host of diseases such as spherocytosis, malaria, and Sickle cell disease, as RBCs depart from their normal discoid shape and lose their deformability. Therefore, quantifying the RBC deformability insight into a variety of problems regarding the interplay of cell structure, dynamics, and function. Furthermore, the ability to monitor mechanical properties of RBCs is of vital interest in monitoring disease progression or response to treatment as molecular and pharmaceutical approaches for treatment of chronic diseases. Here, we present the measurements of dynamic membrane fluctuations in live cells using quantitative phase imaging techniques. Measuring both the 3-D refractive index maps and the dynamic phase images of live cells are simultaneously measured, from which dynamic membrane fluctuation and deformability of cells are precisely calculated. We also present its applications to various diseases ranging from sickle cell diseases, babesiosis, and to diabetes.

Optical diffraction tomography (ODT) has been an emerging optical technique for label-free imaging of three-dimensional (3-D) refractive index (RI) distribution of biological samples. ODT employs interferometric microscopy for measuring multiple holograms of samples with various incident angles, from which the Fourier diffraction theorem reconstructs the 3-D RI distribution of samples from retrieved complex optical fields. Since the RI value is linearly proportional to the protein concentration of biological samples where the proportional coefficient is called as refractive index increment (RII), reconstructed 3-D RI tomograms provide precise structural and biochemical information of individual biological samples. Because most proteins have similar RII value, however, ODT has limited molecular specificity, especially for imaging eukaryotic cells having various types of proteins and subcellular organelles. Here, we present an ODT system combined with structured illumination microscopy which can measure the 3-D RI distribution of biological samples as well as 3-D super-resolution fluorescent images in the same optical setup. A digital micromirror device (DMD) controls the incident angle of the illumination beam for tomogram reconstruction, and the same DMD modulates the structured illumination pattern of the excitation beam for super-resolution fluorescent imaging. We first validate the proposed method for simultaneous optical diffraction tomographic imaging and super-resolution fluorescent imaging of fluorescent beads. The proposed method is also exploited for various biological samples.

In this paper, we present the optical characterisations of diabetic red blood cells (RBCs) in a non-invasive manner employing three-dimensional (3-D) quantitative phase imaging. By measuring 3-D refractive index tomograms and 2-D time-series phase images, the morphological (volume, surface area and sphericity), biochemical (haemoglobin concentration and content) and mechanical (membrane fluctuation) parameters were quantitatively retrieved at the individual cell level. With simultaneous measurements of individual cell properties, systematic correlative analyses on retrieved RBC parameters were also performed. Our measurements show there exist no statistically significant alterations in morphological and biochemical parameters of diabetic RBCs, compared to those of healthy (non-diabetic) RBCs. In contrast, membrane deformability of diabetic RBCs is significantly lower than that of healthy, non-diabetic RBCs. Interestingly, non-diabetic RBCs exhibit strong correlations between the elevated glycated haemoglobin in RBC cytoplasm and decreased cell deformability, whereas diabetic RBCs do not show correlations. Our observations strongly support the idea that slow and irreversible glycation of haemoglobin and membrane proteins of RBCs by hyperglycaemia significantly compromises RBC deformability in diabetic patients.

Using a common-path interferometric technique, we measure biomechanical and morphological properties of individual red blood cells in SCD patients as a function of cell density, and investigate the correlation of these biophysical properties with drug intake as well as other clinically measured parameters.

We propose and demonstrate a system for wavefront shaping, which generates optical foci through complex disordered media and achieves an enhancement factor of greater than 100,000. To exploit the 1 megapixel capacity of a digital micro-mirror device and its fast frame rate, we developed a fast and efficient method to handle the heavy matrix algebra computation involved in optimizing the focus. We achieved an average enhancement factor of 101,391 within an optimization time of 73 minutes with amplitude control. This unprecedented enhancement factor may open new possibilities for realistic image projection and the efficient delivery of energy through scattering media. (C) 2017 Optical Society of America

Two-dimensional (2D) nanomaterials, such as graphene-based materials and transition metal dichalcogenide (TMD) nanosheets, are promising materials for biomedical applications owing to their remarkable cytocompatibility and physicochemical properties. On the basis of their potent antibacterial properties, 2D materials have potential as antibacterial films, wherein the 2D nanosheets are immobilized on the surface and the bacteria may contact with the basal planes of 2D nanosheets dominantly rather than contact with the sharp edges of nanosheets. To address these points, in this study, we prepared an effective antibacterial surface consisting of representative 2D materials, i.e., graphene oxide (GO) and molybdenum disulfide (MoS2), formed into nanosheets on a transparent substrate for real device applications. The antimicrobial properties of the GO–MoS2 nanocomposite surface toward the Gram-negative bacteria Escherichia coli were investigated, and the GO–MoS2 nanocomposite exhibited enhanced antimicrobial ef...

Identification of lymphocyte cell types is crucial for understanding their pathophysiologic roles in human diseases. Current methods for discriminating lymphocyte cell types primarily relies on labelling techniques with magnetic beads or fluorescence agents, which take time and have costs for sample preparation and may also have a potential risk of altering cellular functions. Here, we present label-free identification of non-activated lymphocyte subtypes using refractive index tomography. From the measurements of three-dimensional refractive index maps of individual lymphocytes, the morphological and biochemical properties of the lymphocytes are quantitatively retrieved. Machine learning methods establish an optimized classification model using the retrieved quantitative characteristics of the lymphocytes to identify lymphocyte subtypes at the individual cell level. We show that our approach enables label-free identification of three lymphocyte cell types (B, CD4+ T, and CD8+ T lymphocytes) with high specificity and sensitivity. The present method will be a versatile tool for investigating the pathophysiological roles of lymphocytes in various diseases including cancers, autoimmune diseases, and virus infections.

Parkinson's disease (PD) is a common neurodegenerative disease. However, therapeutic methods of PD are still limited due to complex pathophysiology in PD. Here, optical measurements of individual neurons from in vitro PD model using optical diffraction tomography (ODT) are presented. By measuring 3D refractive index distribution of neurons, morphological and biophysical alterations in in-vitro PD model are quantitatively investigated. It was found that neurons show apoptotic features in early PD progression. The present approach will open up new opportunities for quantitative investigation of the pathophysiology of various neurodegenerative diseases. © 2017 International Society for Advancement of Cytometry.

Identification of lymphocyte cell types are crucial for understanding their pathophysiological roles in human diseases. Current methods for discriminating lymphocyte cell types primarily rely on labelling techniques with magnetic beads or fluorescence agents, which take time and have costs for sample preparation and may also have a potential risk of altering cellular functions. Here, we present the identification of non-activated lymphocyte cell types at the single-cell level using refractive index (RI) tomography and machine learning. From the measurements of three-dimensional RI maps of individual lymphocytes, the morphological and biochemical properties of the cells are quantitatively retrieved. To construct cell type classification models, various statistical classification algorithms are compared, and the k-NN (k = 4) algorithm was selected. The algorithm combines multiple quantitative characteristics of the lymphocyte to construct the cell type classifiers. After optimizing the feature sets via cross-validation, the trained classifiers enable identification of three lymphocyte cell types (B, CD4+ T, and CD8+ T cells) with high sensitivity and specificity. The present method, which combines RI tomography and machine learning for the first time to our knowledge, could be a versatile tool for investigating the pathophysiological roles of lymphocytes in various diseases including cancers, autoimmune diseases, and virus infections.

We present optical characterizations of live amphibian erythrocytes by utilizing quantitative phase imaging. Without any labeling or preparation procedures, 3-D refractive index tomogram and 2-D phase delay of amphibian erythrocyte were measured using Mach-Zehnder interferometry. Refractive index value in 3-D tomograms represent structural and chemical features of an erythrocyte and the nucleus. In addition, membrane fluctuation of amphibian erythrocytes was measured to be more profound than that of human erythrocytes.

We employed the prior information of the discrete and homogeneous sample features to reconstruct the binary refractive index distribution. This ODT-DART algorithm showed the high-quality reconstruction of a silica bead in the simulation and experiments.

One of the fundamental limitations in photonics is the lack of a bidirectional transducer that can convert optical information into electronic signals or vice versa. In acoustics or at microwave frequencies, wave signals can be simultaneously measured and modulated by a single transducer. In optics, however, optical fields are generally measured via reference-based interferometry or holography using silicone-based image sensors, whereas they are modulated using spatial light modulators. Here, we propose a scheme for an optical bidirectional transducer using a spatial light modulator. By exploiting the principle of time-reversal symmetry of light scattering, two-dimensional reference-free measurement and modulation of optical fields are realized. We experimentally demonstrate the optical bidirectional transducer for optical information consisting of 128 x 128 spatial modes at visible and short-wave infrared wavelengths.

We present a method to measure the vector-field light scattering of individual microscopic objects. The polarization-dependent optical field images are measured with quantitative phase imaging at the sample plane, and then numerically propagated to the far-field plane. This approach allows the two-dimensional polarization-dependent angle-resolved light scattered patterns from individual object to be obtained with high precision and sensitivity. Using this method, we present the measurements of the polarization-dependent light scattering of a liquid crystal droplet and individual silver nanowires over scattering angles of 50{\deg}. In addition, the spectroscopic extension of the polarization-dependent angle-resolved light scattering is demonstrated using wavelength-scanning illumination.

Due to its time-reversal nature, optical phase conjugation generates a monochromatic light wave which retraces its propagation paths. Here, we demonstrate the regeneration of a subwavelength optical focus by phase conjugation. Monochromatic light from a subwavelength source is scattered by random nanoparticles, and the scattered light is phase conjugated at the far-field region by coupling its wavefront into a single-mode optical reflector using a spatial light modulator. Then the conjugated beam retraces its propagation paths and forms a refocus on the source at the subwavelength scale. This is the first direct experimental realisation of subwavelength focusing beyond the diffraction limit with far-field time reversal in the optical domain.

We present a label-free structural characterization of individual pollen grains from trees of genus Pinus using quantitative phase imaging. 3-D refractive index distribution in single pollen grain was measured using optical diffraction tomography. The 3-D structure of pollen grains was addressed without chemical treatment of the sample. Scattering of each pollen grain from actively controlled incident wave was measured to be reconstructed into a 3-D refractive index tomogram. Structures of Pinus pollen grains were evaluated with quantitative features obtained from the refractive index tomograms.

We demonstrate polarization-sensitive measurements of the angle-resolved light scattering from a liquid crystal droplet and silver nano wire utilizing quantitative phase imaging and Fourier transform light scattering.

We present white light quantitative phase imaging unit (WQIU) as a practical realization of quantitative phase imaging. WQPIU which consists of a liquid crystal retarder, birefringent crystals and polarizers, enables quantitative phase imaging at conventional microscopes with incoherent illumination

Optical measurements of the morphological and biochemical imaging of phytoplankton are presented. Employing quantitative phase imaging techniques, 3-D refractive index maps and high-resolution 2-D quantitative phase images of individual live phytoplankton are simultaneously obtained without exogenous labeling agents. In addition, biochemical information of individual phytoplankton including volume, mass, and density of individual phytoplankton are also quantitatively obtained from the measured refractive index distributions. We expect the present method to become a powerful tool for the study of phytoplankton.

Modern optical imaging possesses a huge information capacity whose corresponding space-bandwidth product (SBP) reaches tens of megapixels. However, despite the advances in optical and electronic devices, the SBP of an optical microscope is greatly limited, resulting in a reduced field of view or resolution of an image. In this paper, we exploit the Kramers–Kronig relations in digital holography to achieve high SBP imaging, demonstrating a complex amplitude image that can surpass the SBP of a bright-field image. The capability of the proposed method is demonstrated by imaging static samples and biological tissues. We successfully measure a 4.2-megapixel complex amplitude image whose bright-field counterpart exhibits 16.7 megapixels.

We present a novel spectroscopic quantitative phase imaging technique with a wavelength swept-source, referred to as swept-source diffraction phase microscopy (ssDPM), for quantifying the optical dispersion of microscopic individual samples. Employing the swept-source and the principle of common-path interferometry, ssDPM measures the multispectral full-field quantitative phase imaging and spectroscopic microrefractometry of transparent microscopic samples in the visible spectrum with a wavelength range of 450-750 nm and a spectral resolution of less than 8 nm. With unprecedented precision and sensitivity, we demonstrate the quantitative spectroscopic microrefractometry of individual polystyrene beads, 30% bovine serum albumin solution, and healthy human red blood cells.

We propose textured plastic films attached to the external surface of the substrates of organic photovoltaic devices (OPVs) as spectrally neutral light trapping schemes. Both randomly textured films and those containing an array of V-grooves are explored from experimental and theoretical perspectives. Randomly textured surface, providing a close approximation to the Lambertian scatterer that increases the total optical path length of the incident light, is shown to yield an enhancement in short-circuit current density (Jsc) as large as 9.3% in polymer:fullerene bulk heterojunction cells. Different from the case of random texturing, V-groove texturing scheme manages the light in a controlled manner yet can still enhance the optical path length upon careful optimization. In particular, a V-groove with the vertex angle of 100.8–112.0° is found to be highly effective as it suppresses the internal rays from escaping the structure. Together with use of highly reflecting Ag electrodes, V-grooves with the optimal vertex angle leads to 15% enhancement in Jsc, improving the efficiency of OPV cells under study from 6.5% to 7.4%.

Focusing beyond the diffraction limit is achieved by using elastic light scattering from a highly turbid medium to convert propagating far-field components into near-field wave vectors. This finding may open new avenues for the subwavelength control of light, with applications in nanolithography and the interconnection between nanoelectronics and nanophotonics.

A cellular-level study of the pathophysiology is crucial for understanding the mechanisms behind human diseases. Recent advances in quantitative phase imaging (QPI) techniques show promises for the cellular-level understanding of the pathophysiology of diseases. To provide important insight on how the QPI techniques potentially improve the study of cell pathophysiology, here we present the principles of QPI and highlight some of the recent applications of QPI ranging from cell homeostasis to infectious diseases and cancer.

Delivery of gold nanoparticles (GNPs) into live cells has high potentials, ranging from molecular-specific imaging, photodiagnostics, to photothermal therapy. However, studying the long-term dynamics of cells with GNPs using conventional fluorescence techniques suffers from phototoxicity and photobleaching. Here, we present a method for 3-D imaging of GNPs inside live cells exploiting refractive index (RI) as imaging contrast. Employing optical diffraction tomography, 3-D RI tomograms of live cells with GNPs are precisely measured for an extended period with sub-micrometer resolution. The locations and contents of GNPs in live cells are precisely addressed and quantified due to their distinctly high RI values, which was validated by confocal fluorescence imaging of fluorescent dye conjugated GNPs. In addition, we perform quantitative imaging analysis including the segmentations of GNPs in the cytosol, the volume distributions of aggregated GNPs, and the temporal evolution of GNPs contents in HeLa and 4T1 cells.AbbreviationsGNPsgold nanoparticlesRIrefractive indexODToptical diffraction tomographyDMDdigital micromirror device

Optical trapping can manipulate the three-dimensional (3D) motion of spherical particles based on the simple prediction of optical forces and the responding motion of samples. However, controlling the 3D behaviour of non-spherical particles with arbitrary orientations is extremely challenging, due to experimental difficulties and extensive computations. Here, we achieve the real-time optical control of arbitrarily shaped particles by combining the wavefront shaping of a trapping beam and measurements of the 3D refractive index distribution of samples. Engineering the 3D light field distribution of a trapping beam based on the measured 3D refractive index map of samples generates a light mould, which can manipulate colloidal and biological samples with arbitrary orientations and/or shapes. The present method provides stable control of the orientation and assembly of arbitrarily shaped particles without knowing a priori information about the sample geometry. The proposed method can be directly applied in biophotonics and soft matter physics., Controlling the three-dimensional behaviour of arbitrarily shaped and oriented particles with optical tweezers is a challenging task. Here, Kim and Park use tomographic active trapping to manipulate non-spherical particles and particle ensembles as well as biological cells.

Parkinson’s disease (PD) is a common neurodegenerative disease. However, therapeutic methods of PD are still limited due to complex pathophysiology in PD. Here, we present optical measurements of individual neurons fromin vitroPD model using optical diffraction tomography (ODT). By measuring 3-D refractive index distribution of neurons, morphological and biochemical alterations inin-vitroPD model are quantitatively investigated. We found that neurons show apoptotic features in early PD progression. The present approach will open up new opportunities for quantitative investigation of the pathophysiology of various neurodegenerative diseases.

We demonstrate that simultaneous application of optical clearing agents (OCAs) and complex wavefront shaping in optical coherence tomography (OCT) can provide significant enhancement of penetration depth and imaging quality. OCA reduces optical inhomogeneity of a highly scattering sample, and the wavefront shaping of illumination light controls multiple scattering, resulting in an enhancement of the penetration depth and signal-to-noise ratio. A tissue phantom study shows that concurrent applications of OCA and wavefront shaping successfully operate in OCT imaging. The penetration depth enhancement is further demonstrated for

The functionality and viability of stored human red blood cells (RBCs) is an important clinical issue in transfusions. To systematically investigate changes in stored whole blood, the hematological properties of individual RBCs were quantified in blood samples stored for various periods with and without a preservation solution called citrate phosphate dextrose adenine-1 (CPDA-1). With 3-D quantitative phase imaging techniques, the optical measurements for 3-D refractive index (RI) distributions and membrane fluctuations were done at the individual cell level. From the optical measurements, the morphological (volume, surface area and sphericity), biochemical (hemoglobin content and concentration), and mechanical parameters (dynamic membrane fluctuation) were simultaneously quantified to investigate the functionalities and progressive alterations of stored RBCs. Our results show that stored RBCs without CPDA-1 had a dramatic morphological transformation from discocytes to spherocytes within two weeks which was accompanied by significant decreases in cell deformability and cell surface area, and increases in sphericity. However, the stored RBCs with CPDA-1 maintained their morphology and deformability for up to 6 weeks.

The word ‘holography' means a drawing that contains all of the information for light—both amplitude and wavefront. However, because of the insufficient bandwidth of current electronics, the direct measurement of the wavefront of light has not yet been achieved. Though reference-field-assisted interferometric methods have been utilized in numerous applications, introducing a reference field raises several fundamental and practical issues. Here we demonstrate a reference-free holographic image sensor. To achieve this, we propose a speckle-correlation scattering matrix approach; light-field information passing through a thin disordered layer is recorded and retrieved from a single-shot recording of speckle intensity patterns. Self-interference via diffusive scattering enables access to impinging light-field information, when light transport in the diffusive layer is precisely calibrated. As a proof-of-concept, we demonstrate direct holographic measurements of three-dimensional optical fields using a compact device consisting of a regular image sensor and a diffusor., Holographic techniques store and retrieve complete optical information, but the requirement of a reference beam can make the process complicated and sensitive to noise. Here, the authors develop a reference-free method that harnesses self-interference in a diffusive scattering medium.

Sickle cell disease (SCD) is common across Sub-Saharan Africa. However, the investigation of SCD in this area has been significantly limited mainly due to the lack of research facilities and skilled personnel. Here, we present optical measurements of individual red blood cells from healthy individuals and individuals with SCD and sickle cell trait in Tanzania using the quantitative phase imaging technique. By employing a quantitative phase imaging unit, an existing microscope in a clinic is transformed into a powerful quantitative phase microscope providing measurements on the morphological, biochemical, and biomechanical properties of individual cells. The present approach will open up new opportunities for cost-effective investigation and diagnosis of several diseases in low resource environments.

Hydroxyurea (HU) has been used clinically to reduce the frequency of painful crisis and the need for blood transfusion in sickle cell disease (SCD) patients. However, the mechanisms underlying such beneficial effects of HU treatment are still not fully understood. Studies have indicated a weak correlation between clinical outcome and molecular markers, and the scientific quest to develop companion biophysical markers have mostly targeted studies of blood properties under hypoxia. Using a common-path interferometric technique, we measure biomechanical and morphological properties of individual red blood cells in SCD patients as a function of cell density, and investigate the correlation of these biophysical properties with drug intake as well as other clinically measured parameters. Our results show that patient-specific HU effects on the cellular biophysical properties are detectable at normoxia, and that these properties are strongly correlated with the clinically measured mean cellular volume rather than fetal hemoglobin level.

We present a wide-field quantitative label-free imaging of mouse brain tissue slices with sub-micrometre resolution, employing holographic microscopy and an automated scanning platform. From the measured light field images, scattering coefficients and anisotropies are quantitatively retrieved by using the modified the scattering-phase theorem, which enables access to structural information about brain tissues. As a proof of principle, we demonstrate that these scattering parameters enable us to quantitatively address structural alteration in the brain tissues of mice with Alzheimer’s disease.

Three-dimensional imaging of biological cells is crucial for the investigation of cell biology, providing valuable information to reveal the mechanisms behind pathophysiology of cells and tissues. Recent advances in optical diffraction tomography (ODT) have demonstrated the potential for the study of various cells with its unique advantages of quantitative and label-free imaging capability. To provide insight on this rapidly growing field of research and to discuss its applications in biology and medicine, we present the summary of the ODT principle and highlight recent studies utilizing ODT with the emphasis on the applications to the pathophysiology of cells.

When a light propagates through highly disordered medium, its optical parameters such as amplitude, phase and polarization states are completely scrambled because of multiple scattering events. Since the multiple scattering is a fundamental optical process that contains extremely high degrees of freedom, optical information of a transmitted light is totally mingled. Until recently, the presence of multiple scattering in an inhomogeneous medium is considered as a major obstacle when manipulating a light transmitting through the medium. However, a recent development of wavefront shaping techniques enable us to control the propagation of light through turbid media; a light transmitting through a turbid medium can be effectively controlled by modulating the spatial profile of the incident light using spatial light modulator. In this work, stand-alone scattering optical device is proposed; a holographic photopolymer film, which is much economic compared to the other digital spatial light modulators, is used to record and reconstruct permanent wavefront to generate optical field behind a scattering medium. By employing our method, arbitrary optical field can be generated since the scattering medium completely mixes all the optical parameters which allow us to access all the optical information only by modulating spatial phase profile of the impinging wavefront. The method is experimentally demonstrated in both the far-field and near-field regime where it shows promising fidelity and stability. The proposed stand-alone scattering optical device will opens up new avenues for exploiting the randomness inherent in disordered medium.

White blood cells (WBC) have crucial roles in immune systems which defend the host against from disease conditions and harmful invaders. Various WBC subsets have been characterized and reported to be involved in many pathophysiologic conditions. It is crucial to isolate a specific WBC subset to study its pathophysiological roles in diseases. Identification methods for a specific WBC population are rely on invasive approaches, including Wright-Gimesa staining for observing cellular morphologies and fluorescence staining for specific protein markers. While these methods enable precise classification of WBC populations, they could disturb cellular viability or functions. In order to classify WBC populations in a non-invasive manner, we exploited optical diffraction tomography (ODT). ODT is a three-dimensional (3-D) quantitative phase imaging technique that measures 3-D refractive index (RI) distributions of individual WBCs. To test feasibility of label-free classification of WBC populations using ODT, we measured four subtypes of WBCs, including B cell, CD4 T cell, CD8 T cell, and natural killer (NK) cell. From measured 3-D RI tomograms of WBCs, we obtain quantitative structural and biochemical information and classify each WBC population using a machine learning algorithm.