Scattering-Based Light-Sheet Microscopy Imaging of Human Papillomavirus-Associated Squamous Lesions of the Anal Canal: A Proof-of-Principle Study.

Demand for anal cancer screening is expected to rise following the recent publication of the Anal Cancer-HSIL Outcomes Research trial, which showed that treatment of high-grade squamous intraepithelial lesions significantly reduces the rate of progression to anal cancer. While screening for human papillomavirus-associated squamous lesions in the cervix is well established and effective, this is less true for other sites in the lower anogenital tract. Current anal cancer screening and prevention rely on high-resolution anoscopy with biopsies. This procedure has a steep learning curve for providers and may cause patient discomfort. Scattering-based light-sheet microscopy (sLSM) is a novel imaging modality with the potential to mitigate these challenges through real-time, microscopic visualization of disease-susceptible tissue. Here, we report a proof-of-principle study that establishes feasibility of dysplasia detection using an sLSM device. We imaged 110 anal biopsy specimens collected prospectively at our institution's dysplasia clinic (including 30 nondysplastic, 40 low-grade squamous intraepithelial lesion, and 40 high-grade squamous intraepithelial lesion specimens) and found that these optical images are highly interpretable and accurately recapitulate histopathologic features traditionally used for the diagnosis of human papillomavirus-associated squamous dysplasia. A reader study to assess diagnostic accuracy suggests that sLSM images are noninferior to hematoxylin and eosin images for the detection of anal dysplasia (sLSM accuracy = 0.87; hematoxylin and eosin accuracy = 0.80; P = .066). Given these results, we believe that sLSM technology holds great potential to enhance the efficacy of anal cancer screening by allowing accurate sampling of diagnostic tissue at the time of anoscopy. While the current imaging study was performed on ex vivo biopsy specimens, we are currently developing a handheld device for in vivo imaging that will provide immediate microscopic guidance to high-resolution anoscopy providers.

Laser Dynamic Control of the Thermal Emissivity of a Planar Cavity Structure Based on a Phase-Change Material.

Tailoring the thermal emission of a material in the long-wave infrared (IR) range of 8-13 µm is crucial for many IR-adaptive applications, including personal thermal management, IR camouflage, and radiative cooling. Although various materials and surface structures have been proposed for these purposes, space-selective and dynamic control of their emissivity is challenging. In this study, we present a planar surface cavity structure consisting of a Ge2Sb2Te5 (GST) film on top of a thin metal reflector to modulate its emissivity by using an ultraviolet laser beam. A laser-induced phase change in GST allowed for the local control of emissivity. The average emissivity in the long-wave IR range was tunable from 0.15 to 0.77 simply by changing the laser energy deposited on the GST film. This enabled the laser printing of high-contrast emissivity patterns, which were erasable by subsequent thermal annealing. Emissivity-modulated GST cavities could be fabricated on not only rigid substrates but also flexible plastic substrates such as polyimide. The GST surface cavity was highly flexible and remained stable upon repeated bending to a curvature radius of 0.5 cm. This study provides a promising route for realizing scalable and flexible thermal emitters with tunable surface emissivity.

Handheld cross-polarised microscope for imaging individual pigmented cells in human skin in vivo.

We present the development of a simple, handheld cross-polarised microscope (CPM) and demonstration of imaging individual pigmented cells in human skin in vivo. In the CPM device, the cross-polarised detection approach is used to reduce the specular reflection from the skin surface and preferentially detect multiply-scattered light. The multiply-scattered light works as back illumination from within the tissue towards the skin surface, and superficial pigment such as intraepidermal melanin absorbs some spectral bands of the multiply-scattered light and cast coloured shadows. Since the light that interacted with the superficial pigment only needs to travel a short distance before it exits the skin surface, microscopic details of the pigment can be preserved. The CPM device uses a water-immersion objective lens with a high numerical aperture to image the microscopic details with minimal spherical aberrations and a small depth of focus. Preliminary results from a pilot study of imaging skin lesions in vivo showed that the CPM device could reveal three-dimensional distribution of pigmented cells and intracellular distribution of pigment. Co-registered CPM and reflectance confocal microscopy images showed good correspondence between dark, brown cells in CPM images and bright, melanin-containing cells in reflectance confocal microscopy images.

Titanium oxide nanomaterials as an electron-selective contact in silicon solar cells for photovoltaic devices.

To obtain high conversion efficiency, various carrier-selective contact structures are being applied to the silicon solar cell, and many related studies are being conducted. We conducted research on TiO2 to create an electron-selective contact structure that does not require a high-temperature process. Titanium metal was deposited using a thermal evaporator, and an additional oxidation process was conducted to form titanium oxide. The chemical compositions and phases of the titanium dioxide layers were analyzed by X-ray diffraction. The passivation effects of each titanium oxide layer were measured using the quasi-steady-state photoconductance. In this study, the layer properties were analyzed when TiO2 had a passivation effect on the silicon surface. The charge and interface defect densities of the layer were analyzed through CV measurements, and the passivation characteristics according to the TiO2 phase change were investigated. As a result, by applying optimized TiO2 layer thickness and annealing temperature conditions through the experiment for passivation to the cell-like structure, which is the structure before metal and electrode formation, an implied open-circuit voltage (iVoc) of 630 mV and an emitter saturation current density (J0) value of 60.4 fA/cm2 were confirmed.

Multispectral Imaging with a Planar Cavity-Type Metasurface for Optical Security.

Multispectral imaging refers to capturing images in different wavelength ranges across the electromagnetic spectrum. Despite the potential impact of multispectral imaging, its widespread use has been limited by the poor spectral selectivity of naturally occurring materials beyond the visible range. In this study, we present a multilayered planar cavity structure to simultaneously record mutually independent visible and infrared (IR) images on solid surfaces. The structure consists of a color control unit (CCU) and an emission control unit (ECU). The visible color of the cavity is controlled by varying the thickness of the CCU, whereas its IR emission is spatially tuned by the laser-induced phase change of a Ge2Sb2Te5 layer embedded in the ECU. Because the CCU comprises only IR lossless layers, its thickness variation has negligible influence on the emission profile. This enables different color and thermal images to be printed in a single structure. The cavity structure can be fabricated on flexible substrates (plastic and paper) as well as rigid bodies. Furthermore, the printed images remain stable against bending. This study shows that the proposed multispectral metasurface is highly promising for use in the field of optical security, such as identification, authentication, and anti-counterfeiting.

High-speed reflectance confocal microscopy of human skin at 1251-1342 nm.

OBJECTIVES: Reflectance confocal microscopy (RCM) is an imaging method that can noninvasively visualize microscopic features of the human skin. The utility of RCM can be further improved by increasing imaging speed. In this paper, we report high-speed RCM imaging of human skin with a frame rate that is over 10 times faster and an area imaging rate that is 6-9 times faster than those of commercially available RCM devices. METHODS: The higher imaging speed was achieved using a high-speed RCM technique, termed spectrally encoded confocal microscopy (SECM). SECM uses a diffraction grating and a high-speed, wavelength-swept source to conduct confocal imaging at a very high rate. We developed a handheld SECM probe using a scanned-grating approach. The SECM probe was used in conjunction with a wavelength-swept source with a spectral band of 1251-1342 nm. RESULTS: The SECM probe achieved high lateral resolution of 1.3-1.6 µm and an axial resolution of 3.5 µm. SECM images of the human skin (image size = 439 × 439 µm2 ) obtained at 100 frames/s clearly show previously reported RCM features of the human skin in vivo with adequate image quality. The fast imaging speed allowed for the rapid acquisiton of volumetric SECM image data (200 frames covering a depth range of 200 µm) within 2 s. The use of 1251-1342 nm provided sufficient signal level and contrast required to visualize key cellular morphologic features. CONCLUSIONS: These preliminary results demonstrate that high-speed SECM imaging of the human skin at 1251-1342 nm is feasible.

Investigation of different wavelengths for scattering-based light sheet microscopy.

Scattering-based light sheet microscopy (sLSM) is a microscopy technique that can visualize cellular morphologic details based on the scattering signal. While sLSM was previously shown to image animal tissues ex vivo at a cellular resolution, the wavelength used was chosen based on other in vivo microscopy technologies rather than through a comparison of the sLSM imaging performance between different wavelengths. In this paper, we report the development of a multi-wavelength sLSM setup that facilitates the investigation of different wavelengths for sLSM imaging. Preliminary results of imaging human anal tissues ex vivo showed that the sLSM setup allowed for comparisons of the cellular imaging performance at the same tissue location between different wavelengths. Both the quantitative analysis of the image contrast and the visual assessment by a pathologist showed that the imaging depth increased with wavelength, and the imaging depth increase was most notable around 600 nm. The preliminary results showed that the multi-wavelength sLSM setup could be useful in identifying the optimal wavelength for the specific tissue type.

Low-cost, chromatic confocal endomicroscope for cellular imaging in vivo.

We have developed a low-cost, chromatic confocal endomicroscope (CCE) that can image a cross-section of the tissue at cellular resolution. In CCE, a custom miniature objective lens was used to focus different wavelengths into different tissue depths. Therefore, each tissue depth was encoded with the wavelength. A custom miniature spectrometer was used to spectrally-disperse light reflected from the tissue and generate cross-sectional confocal images. The CCE prototype had a diameter of 9.5 mm and a length of 68 mm. Measured resolution was high, 2 µm and 4 µm for lateral and axial directions, respectively. Effective field size was 468 µm. Preliminary results showed that CCE can visualize cellular details from cross-sections of the tissue in vivo down to the tissue depth of 100 µm.

Imaging the dynamics and microstructure of fibrin clot polymerization in cardiac surgical patients using spectrally encoded confocal microscopy.

During cardiac surgery with cardiopulmonary bypass (CPB), altered hemostatic balance may disrupt fibrin assembly, predisposing patients to perioperative hemorrhage. We investigated the utility of a novel device termed spectrally-encoded confocal microscopy (SECM) for assessing fibrin clot polymerization following heparin and protamine administration in CPB patients. SECM is a novel, high-speed optical approach to visualize and quantify fibrin clot formation in three dimensions with high spatial resolution (1.0 µm) over a volumetric field-of-view (165 × 4000 × 36 µm). The measurement sensitivity of SECM was first determined using plasma samples from normal subjects spiked with heparin and protamine. Next, SECM was performed in plasma samples from patients on CPB to quantify the extent to which fibrin clot dynamics and microstructure were altered by CPB exposure. In spiked samples, prolonged fibrin time (4.4 ± 1.8 to 49.3 ± 16.8 min, p < 0.001) and diminished fibrin network density (0.079 ± 0.010 to 0.001 ± 0.002 A.U, p < 0.001) with increasing heparin concentration were reported by SECM. Furthermore, fibrin network density was not restored to baseline levels in protamine-treated samples. In CPB patients, SECM reported lower fibrin network density in protaminized samples (0.055 ± 0.01 A.U. [Arbitrary units]) vs baseline values (0.066 ± 0.009 A.U.) (p = 0.03) despite comparable fibrin time (baseline = 6.0 ± 1.3, protamine = 6.4 ± 1.6 min, p = 0.5). In these patients, additional metrics including fibrin heterogeneity, length and straightness were quantified. Note, SECM revealed that following protamine administration with CPB exposure, fibrin clots were more heterogeneous (baseline = 0.11 ± 0.02 A.U, protamine = 0.08 ± 0.01 A.U, p = 0.008) with straighter fibers (baseline = 0.918 ± 0.003A.U, protamine = 0.928 ± 0.0006A.U. p < 0.001). By providing the capability to rapidly visualize and quantify fibrin clot microstructure, SECM could furnish a new approach for assessing clot stability and hemostasis in cardiac surgical patients.

Deep Learning-Based Denoising in High-Speed Portable Reflectance Confocal Microscopy.

BACKGROUND AND OBJECTIVE: Portable confocal microscopy (PCM) is a low-cost reflectance confocal microscopy technique that can visualize cellular details of human skin in vivo. When PCM images are acquired with a short exposure time to reduce motion blur and enable real-time 3D imaging, the signal-to-noise ratio (SNR) is decreased significantly, which poses challenges in reliably analyzing cellular features. In this paper, we evaluated deep learning (DL)-based approach for reducing noise in PCM images acquired with a short exposure time. STUDY DESIGN/MATERIALS AND METHODS: Content-aware image restoration (CARE) network was trained with pairs of low-SNR input and high-SNR ground truth PCM images obtained from 309 distinctive regions of interest (ROIs). Low-SNR input images were acquired from human skin in vivo at the imaging speed of 180 frames/second. The high-SNR ground truth images were generated by registering 30 low-SNR input images obtained from the same ROI and summing them. The CARE network was trained using the Google Colaboratory Pro platform. The denoising performance of the trained CARE network was quantitatively and qualitatively evaluated by using image pairs from 45 unseen ROIs. RESULTS: CARE denoising improved the image quality significantly, increasing similarity with the ground truth image by 1.9 times, reducing noise by 2.35 times, and increasing SNR by 7.4 dB. Banding noise, prominent in input images, was significantly reduced in CARE denoised images. CARE denoising provided quantitatively and qualitatively better noise reduction than non-DL filtering methods. Qualitative image assessment by three confocal readers showed that CARE denoised images exhibited negligible noise more often than input images and non-DL filtered images. CONCLUSIONS: Results showed the potential of using a DL-based method for denoising PCM images obtained at a high imaging speed. The DL-based denoising method needs to be further trained and tested for PCM images obtained from disease-suspicious skin lesions.

Scattering-Based Light-Sheet Microscopy for Rapid Cellular Imaging of Fresh Tissue.

BACKGROUND AND OBJECTIVES: Light-sheet microscopy (LSM) is a novel imaging technology that has been used for imaging fluorescence contrast in basic life science research. In this paper, we have developed a scattering-based LSM (sLSM) for rapidly imaging the cellular morphology of fresh tissues without any exogenous fluorescent dyes. STUDY DESIGN/MATERIALS AND METHODS: In the sLSM device, a thin light sheet with the central wavelength of 834 nm was incident on the tissue obliquely, 45° relative to the tissue surface. The detection optics was configured to map the light sheet-illuminated area onto a two-dimensional imaging sensor. The illumination numerical aperture (NA) was set as 0.0625, and the detection NA 0.3. RESULTS: The sLSM device achieved a light sheet thickness of less than 6.7 µm over 284 µm along the illumination optical axis. The detection optics of the sLSM device had a resolution of 1.8 µm. The sLSM images of the swine kidney ex vivo visualized tubules with similar sizes and shapes to those observed in histopathologic images. The swine duodenum sLSM images revealed cell nuclei and villi architecture in superficial lesions and glands in deeper regions. CONCLUSIONS: The preliminary results suggest that sLSM may have the potential for rapidly examining the freshly-excised tissue ex vivo or intact tissue in vivo at microscopic resolution. Further optimization and performance evaluation of the sLSM technology will be needed in the future. Lasers Surg. Med. © 2020 Wiley Periodicals LLC.

Novel Diagnostics for Kaposi Sarcoma and Other Skin Diseases in Resource-Limited Settings.

In resource-limited settings, point-of-care diagnostic devices have the potential to reduce diagnostic delays and improve epidemiologic surveillance of dermatologic conditions. We outline novel-point-of care diagnostics that have recently been developed for dermatologic conditions that primarily affect patients living in resource-limited settings, namely, Kaposi sarcoma, cutaneous leishmaniasis, leprosy, Buruli ulcer, yaws, onchocerciasis, and lymphatic filariasis. All of the technologies described in this article are prototypes, and some have undergone field testing. These devices still require validation in real-world settings and effective pricing to have a major impact on dermatologic care in resource-limited settings.

Speckle-free, near-infrared portable confocal microscope.

We have developed a portable confocal microscope (PCM) that uses an inexpensive near-infrared LED as the light source. Use of the spatially incoherent light source significantly reduced the speckle contrast. The PCM device was manufactured at the material cost of approximately $5000 and weighed only 1 kg. Lateral and axial resolutions were measured as 1.6 and 6.0 µm, respectively. Preliminary in vivo skin imaging experiment results showed that the PCM device could visualize characteristic cellular features of human skin extending from the stratum corneum to the superficial dermis. Dynamic imaging of blood flow in vivo was also demonstrated. The capability to visualize cellular features up to the superficial dermis is expected to facilitate evaluation and clinical adoption of this low-cost diagnostic imaging tool.

Smartphone epifluorescence microscopy for cellular imaging of fresh tissue in low-resource settings.

Disease diagnosis in low-resource settings can be challenging due to the lack of equipment and trained personnel required for histologic analysis. In this paper, we have developed a smartphone-based epifluorescence microscope (SeFM) for imaging fresh tissues at sub-cellular resolution. SeFM provides similar resolution and field of view (FOV) as those used during histologic analysis. The SeFM device achieved the lateral resolution of 0.57 µm and provided microscopy images over a sample area larger than 500 µm. The material cost was low, approximately $3,000. Preliminary images of human pancreatic tumor specimens clearly visualized cellular details. Quantitative analysis showed that using an excess dose of a chemotherapy drug significantly reduced the tumor-specific fluorescence signal, confirming the specificity of the drug and the detection potential of SeFM.

Perovskites fabricated on textured silicon surfaces for tandem solar cells.

The silicon surface texture significantly affects the current density and efficiency of perovskite/silicon tandem solar cells. However, only a few studies have explored fabricating perovskite on textured silicon and the effect of texture on perovskite films because of the limitations of solution processes. Here we produce conformal perovskite on textured silicon with a dry two-step conversion process that incorporates lead oxide sputtering and direct contact with methyl ammonium iodide. To separately analyze the influence of each texture structure on perovskite films, patterned texture, high-resolution photoluminescence (µ-PL), and light beam-induced current (µ-LBIC), 3D mapping is used. This work elucidates conformal perovskite on textured surfaces and shows the effects of textured silicon on the perovskite layers with high-resolution 3D mapping. This approach can potentially be applied to any type of layer on any type of substrate.