Moosung Lee

Label-free optical quantification of structural alterations in Alzheimer's disease.

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.

Label-free non-invasive quantitative measurement of lipid contents in individual microalgal cells using refractive index tomography

Microalgae are promising candidates for biofuel production due to their high lipid content. To facilitate utilization of the microalgae for biofuel, rapid quantification of the lipid contents in microalgae is necessary. However, conventional methods based on the chemical extraction of lipids require a time-consuming destructive extraction process. Here, we demonstrate label-free, non-invasive, rapid quantification of the lipid contents in individual micro-algal cells measuring the three-dimensional refractive index tomograms. We measure three-dimensional refractive index distributions within Nannochloropsis oculata cells and find that lipid droplets are identifiable in tomograms by their high refractive index. In addition, we alter N. oculata under nitrogen deficiency by measuring the volume, lipid weight, and dry cell weight of individual cells. Characterization of individual cells allows correlative analysis between the lipid content and size of individual cells.

Refractive index as an intrinsic imaging contrast for 3-D label-free live cell imaging

Live cell imaging provides critical information in the investigation of cell biology and related pathophysiology. Refractive index (RI) can serve as intrinsic optical imaging contrast for 3-D label-free and quantitative live cell imaging, and provide invaluable information to understand various dynamics of cells and tissues for the study of numerous fields. Recently significant advances have been made in imaging methods and analysis approaches utilizing RI, which are now being transferred to biological and medical research fields, providing novel approaches to investigate the pathophysiology of cells. To provide insight how RI can be used as an imaging contrast for bioimaging, here we provide the basic principle of RI-based imaging techniques and summarize recent progress on applications, ranging from microbiology, hematology, infectious diseases, hematology, and histopathology. Impact Statement The use of refractive index has been utilized for label-free and quantitative imaging contrast of cells and tissues. This mini review presents the principles of imaging techniques which exploit refractive index as an intrinsic optical imaging contrast, discusses the advantages and challenges, and summarize recent progress in various fields of biology and medicine.

Reconstructions of refractive index tomograms via a discrete algebraic reconstruction technique

Optical diffraction tomography (ODT) provides three-dimensional refractive index (RI) tomograms of a transparent microscopic object. However, because of the finite numerical aperture of objective lenses, ODT has a limited access to diffracted light and suffers from poor spatial resolution, particularly along the axial direction. To overcome the limitation of the quality of RI tomography, we present an algorithm that accurately reconstructs RI tomography of a specimen with discrete and uniform RI, using prior information about the RI levels. Through simulations and experiments on various samples, including microspheres, red blood cells, and water droplets, we show that the proposed method can precisely reconstruct RI tomograms of samples which have discrete and homogenous RI distributions in the presence of the missing information and noise.

Measurements of three-dimensional refractive index tomography and membrane deformability of live erythrocytes from Pelophylax nigromaculatus

Unlike mammalian erythrocytes, amphibian erythrocytes have distinct morphological features including large cell sizes and the presence of nuclei. The sizes of the cytoplasm and nuclei of erythrocytes vary significantly over different species, their environments, or pathophysiology, which makes hematological studies important for investigating amphibian species. Here, we present a label-free three-dimensional optical quantification of individual amphibian erythrocytes from frogs Pelophylax nigromaculatus (Rana nigromaculata). Using optical diffraction tomography, we measured three-dimensional refractive index (RI) tomograms of the cells, which clearly distinguished the cytoplasm and nuclei of the erythrocytes. From the measured RI tomograms, we extracted the relevant biochemical parameters of the cells, including hemoglobin contents and hemoglobin concentrations. Furthermore, we measured dynamic membrane fluctuations and investigated the mechanical properties of the cell membrane. From the statistical and correlative analysis of these retrieved parameters, we investigated interspecific differences between frogs and previously studied mammals.

Quantifying structural alterations in Alzheimer's disease brains using quantitative phase imaging (Conference Presentation)

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.

Reconstructing binary refractive index tomograms with discrete algebraic reconstruction technique

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.

Identification of amyloid plaques in mouse brain tissue slides using quantitative phase imaging

We demonstrate that quantitative phase imaging (QPI) can detect amyloid plaques in the brain of Alzheimers disease (AD). Comparing QPIs and fluorescence images from wild-type and AD mice brains, we suggest that digital microscopic holography can be utilized for diagnosing AD.