In vivo whole-brain imaging of zebrafish larvae using three-dimensional fluorescence microscopy.

As a vertebrate model animal, larval zebrafish are widely used in neuroscience and provide a unique opportunity to monitor whole-brain activity at the cellular resolution. Here, we provide an optimized protocol for performing whole-brain imaging of larval zebrafish using three-dimensional fluorescence microscopy, including sample preparation and immobilization, sample embedding, image acquisition, and visualization after imaging. The current protocol enables in vivo imaging of the structure and neuronal activity of a larval zebrafish brain at a cellular resolution for over 1 h using confocal microscopy and custom-designed fluorescence microscopy. The critical steps in the protocol are also discussed, including sample mounting and positioning, preventing bubble formation and dust in the agarose gel, and avoiding motion in images caused by incomplete solidification of the agarose gel and paralyzation of the fish. The protocol has been validated and confirmed in multiple settings. This protocol can be easily adapted for imaging other organs of a larval zebrafish.

Progressive compressive sensing of large images with multiscale deep learning reconstruction.

Compressive sensing (CS) is a sub-Nyquist sampling framework that has been employed to improve the performance of numerous imaging applications during the last 15 years. Yet, its application for large and high-resolution imaging remains challenging in terms of the computation and acquisition effort involved. Often, low-resolution imaging is sufficient for most of the considered tasks and only a fraction of cases demand high resolution, but the problem is that the user does not know in advance when high-resolution acquisition is required. To address this, we propose a multiscale progressive CS method for the high-resolution imaging. The progressive sampling refines the resolution of the image, while incorporating the already sampled low-resolution information, making the process highly efficient. Moreover, the multiscale property of the progressively sensed samples is capitalized for a fast, deep learning (DL) reconstruction, otherwise infeasible due to practical limitations of training on high-resolution images. The progressive CS and the multiscale reconstruction method are analyzed numerically and demonstrated experimentally with a single pixel camera imaging system. We demonstrate 4-megapixel size progressive compressive imaging with about half the overall number of samples, more than an order of magnitude faster reconstruction, and improved reconstruction quality compared to alternative conventional CS approaches.

Development of a teledentistry system for the elderly in the context of the covid-19 pandemic: 3d diagnostic models integration in a web platform / Desarrollo de un sistema de teleodontología para adultos mayores en el contexto de la pandemia de covid-19: integración de modelos de diagnóstico 3d en una plataforma web

Objective: To recognize the usefulness of incorporating Three-Dimensional models of standardized humans in electronic health records, in the context of the development of a teledentistry web platform designed for the attention of the elderly population in COVID-19 pandemic context. Material and Methods: A teledentistry web platform designed with different modules for clinical records. Through a new user-computer interface with a standardized virtual 3D phantom, an extraoral physical examination, an intraoral examination section was modeled. A label-associated marker is allowed to record descriptive aspects of the findings. A 3D odontogram represents multiple patient's conditions for each of the 32 dental positions. Results: From a total of 135 patients registered on the platform, 51 markers and 33 photographs associated with the surface of the virtual 3D phantoms were recorded. For the Location parameter: Hard palate 27.6%, inserted gingiva 15.7%, tongue 15.6%. For the Type of lesion parameter (according to the information entered in the pathology selector): unidentified 35.3%, sub-prosthetic stomatitis 23.5%, irritative fibroma 9.8%. Through the registration of the exact location of the finding in the virtual phantom by a 3D marker, the 3D modeling of the oral pathologies contributed to a better diagnosis, improving the remote communication between the attending dentist and specialists. Conclusion: The combination of the 3D modeling and anatomical-referencing in a teledentistry platform can become a powerful tool for the dental practice, due to their utility and specificity.

Objetivo: Reconocer la utilidad de incorporar modelos tridimensionales de humanos estandarizados en registros electrónicos de salud, en el contexto del desarrollo de una plataforma web de teleodontología diseñada para la atención de la población adulta mayor en contexto de pandemia por COVID-19. Material y Métodos: Una plataforma web de teleodontología diseñada con diferentes módulos para historias clínicas. A través de una nueva interfaz usuario-computadora con un fantoma 3D virtual estandarizado, se modeló un examen físico extraoral, una sección de examen intraoral. Se permite un marcador asociado a la etiqueta para registrar aspectos descriptivos de los hallazgos. Un odontograma 3D representa múltiples condiciones del paciente para cada una de las 32 posiciones dentales.Resultados: De un total de 135 pacientes registrados en la plataforma, se registraron 51 marcadores y 33 fotografías asociadas a la superficie de los fantomas virtuales 3D. Para el parámetro Ubicación: Paladar duro 27,6%, encía insertada 15,7%, lengua 15,6%. Para el parámetro Tipo de lesión (según la información ingresada en el selector de patología): no identificado 35,3%, estomatitis subprotésica 23,5%, fibroma irritativo 9,8%. A través del registro de la ubicación exacta del hallazgo en el fantoma virtual mediante un marcador 3D, el modelado 3D de las patologías orales contribuyó a un mejor diagnóstico, mejorando la comunicación remota entre el odontólogo tratante y los especialistas. Conclusión: La combinación del modelado 3D y la referenciación anatómica en una plataforma de teleodontología puede convertirse en una poderosa herramienta para la práctica odontológica, debido a su utilidad y especificidad.

Handheld and Cost-Effective Fourier Lightfield Microscope.

In this work, the design, building, and testing of the most portable, easy-to-build, robust, handheld, and cost-effective Fourier Lightfield Microscope (FLMic) to date is reported. The FLMic is built by means of a surveillance camera lens and additional off-the-shelf optical elements, resulting in a cost-effective FLMic exhibiting all the regular sought features in lightfield microscopy, such as refocusing and gathering 3D information of samples by means of a single-shot approach. The proposed FLMic features reduced dimensions and light weight, which, combined with its low cost, turn the presented FLMic into a strong candidate for in-field application where 3D imaging capabilities are pursued. The use of cost-effective optical elements has a relatively low impact on the optical performance, regarding the figures dictated by the theory, while its price can be at least 100 times lower than that of a regular FLMic. The system operability is tested in both bright-field and fluorescent modes by imaging a resolution target, a honeybee wing, and a knot of dyed cotton fibers.

A high-throughput technique to map cell images to cell positions using a 3D imaging flow cytometer.

We develop a high-throughput technique to relate positions of individual cells to their three-dimensional (3D) imaging features with single-cell resolution. The technique is particularly suitable for nonadherent cells where existing spatial biology methodologies relating cell properties to their positions in a solid tissue do not apply. Our design consists of two parts, as follows: recording 3D cell images at high throughput (500 to 1,000 cells/s) using a custom 3D imaging flow cytometer (3D-IFC) and dispensing cells in a first-in-first-out (FIFO) manner using a robotic cell placement platform (CPP). To prevent errors due to violations of the FIFO principle, we invented a method that uses marker beads and DNA sequencing software to detect errors. Experiments with human cancer cell lines demonstrate the feasibility of mapping 3D side scattering and fluorescent images, as well as two-dimensional (2D) transmission images of cells to their locations on the membrane filter for around 100,000 cells in less than 10 min. While the current work uses our specially designed 3D imaging flow cytometer to produce 3D cell images, our methodology can support other imaging modalities. The technology and method form a bridge between single-cell image analysis and single-cell molecular analysis.

Three-dimensional heads-up surgery in ab-interno trabeculotomy: Image processing-assisted trabeculotomy.

PURPOSE: We compared the visibility and surgeon posture between image-processing-assisted trabeculotomy (IP-LOT) using the NGENUITY® 3D visual system and conventional microsurgery (microscope-assisted trabeculotomy; MS-LOT). METHODS: IP-LOT was performed for five pig eyes. The visibility of the trabecular mesh work was evaluated on images of the trabecular mesh work and the posterior surface of the cornea (Cor) obtained under three different conditions. Images were then analyzed using ImageJ® to measure differences in luminance between the trabecular mesh work and Cor. IP-LOT was also performed for eleven human eyes, and the data were analyzed using the same approach as that used for the pig eyes. The length from the surgeon's abdomen to the operative eye (working distance) during MS-LOT and IP-LOT was measured for 12 different surgeons and compared to evaluate surgeon posture. RESULTS: Image processing significantly increased the difference in luminance between the trabecular mesh work and Cor in both pig and human eyes (p < 0.05). Moreover, the working distance in IP-LOT was significantly shorter than that in MS-LOT (p < 0.05). CONCLUSION: Our findings suggest that the NGENUITY® 3D visual system provides better trabecular mesh work visibility than a normal microscope in conventional surgical methods, and it allows surgeons to operate without moving far from the operative eye.

Development of Planar Illumination Strategies for Solving Mysteries in the Sub-Cellular Realm.

Optical microscopy has vastly expanded the frontiers of structural and functional biology, due to the non-invasive probing of dynamic volumes in vivo. However, traditional widefield microscopy illuminating the entire field of view (FOV) is adversely affected by out-of-focus light scatter. Consequently, standard upright or inverted microscopes are inept in sampling diffraction-limited volumes smaller than the optical system's point spread function (PSF). Over the last few decades, several planar and structured (sinusoidal) illumination modalities have offered unprecedented access to sub-cellular organelles and 4D (3D + time) image acquisition. Furthermore, these optical sectioning systems remain unaffected by the size of biological samples, providing high signal-to-noise (SNR) ratios for objective lenses (OLs) with long working distances (WDs). This review aims to guide biologists regarding planar illumination strategies, capable of harnessing sub-micron spatial resolution with a millimeter depth of penetration.

Avaliação da acurácia dos modelos dentários digitais obtidos para cirurgia ortognática / Assessment of the accuracy of digital dental models obtained for orthognathic surgery

O objetivo deste estudo laboratorial foi avaliar a acurácia dos modelos digitais obtidos por duas técnicas de escaneamento (escâner intraoral - Itero 5d Element® - e escâner de bancada -Straumann ®) de um modelo experimental (Nacional Ossos ®) da arcada superior confeccionada em poliuretano e do modelo de gesso obtido desta arcada. Os pontos A ­ 3mm acima do elemento 17; B ­ 3mm acima do elemento 14; C ­ 3mm acima do elemento 24; D ­ 3mm acima do elemento 27; E ­ cúspide mesiovestibular do elemento 16; F - ponto de contato mais incisal entre os elementos 11 e 21; G - Cúspide mesiovestibular do elemento 26 foram utilizados como referência para as medidas realizadas. As medidas foram realizadas em um software (Geomagic®) de simulação cirúrgica e analisadas quanto a validade e precisão das técnicas de escaneamento utilizadas em todos os grupos (1 a 5). O grupo 1 corresponde ao grupo padrão-ouro; o 2 ao grupo do escaneamento com escâner intraoral do modelo de poliuretano; o 3 ao escaneamento de bancada do modelo; o 4 ao escaneamento com escâner intraoral do modelo de gesso; e 5 ao escaneamento de bancada do modelo de gesso. Na análise da validação, todas as técnicas apresentaram-se válidas quando comparadas ao grupo controle com exceção da medida FG que apresentou diferenças estatisticamente significativas (p<0,05) entre os grupos 1 e 2. A precisão foi avaliada através do índice de correlação intraclasse (CCI) e todas as técnicas apresentaram-se altamente precisas com (CCI) próximo de 1. Desta forma, conclui-se que o escâner intraoral e o escâner de bancada utilizados neste estudo foram confiáveis quando comparados ao grupo controle e que os dois modelos de escâner utilizados se apresentaram com alta precisão (AU).

The objective of this experimental study was to evaluate the accuracy of digital models generated by two scanning techniques (intraoral scanner - Itero 5d Element® - and desktop scanner -Straumann ®) of an experimental model of the upper arch (Nacional Ossos ®) made of polyurethane and the plaster model obtained from this arch. Points A ­ 3mm above element 17; B ­ 3mm above element 14; C ­ 3mm above element 24; D ­ 3mm above element 27; E ­ mesiobuccal cusp of element 16; F - most incisal point of contact between elements 11 and 21; G - Mesiobuccal cusp of element 26 were used as a reference for the measurements performed. The measurements were performed in a surgical simulation software (Geomagic ®) and analyzed for the validity and precision of the scanning techniques used in all groups (1 to 5). Group 1 corresponds to the gold standard group; 2 to the scanning group with intraoral scanner of the polyurethane model; 3 to the desktop scan of the model; 4 to intraoral scanner scanning of the plaster model; and 5 to the desktop scan of the plaster model. In the validation analysis, all techniques showed to be valid compared to the control group, except the FG measure, which showed statistically significant differences (p<0.05) between groups 1 and 2. Precision was assessed using the intraclass correlation(ICC) index, and all techniques were highly accurate with an ICC close to 1. Thus, it is concluded that the intraoral scanner and the bench scanner used in this study were reliable compared to the control group and that the two scanner models used presented themselves with high precision (AU).

The utility of the exoscope compared to the operating microscope in simulated temporal bone surgery.

OBJECTIVE: To objectively assess the utility of an exoscope during simulated otological surgery. DESIGN: Cohort study. SETTING: Tertiary referral otolaryngology centre. PARTICIPANTS: Seven experienced otologists undertook simulated temporal bone surgery on plastic temporal bones using the Zeiss Kinevo microscope with both a microscope and exoscope facility. OUTCOME MEASURES: The utility of microscope and exoscope was compared using a Likert scale from 1 to 10 with and without PPE. Attributes assessed included image quality, depth perception, adequacy of view, exoscope positioning, surgeon comfort, surgeon safety and adequacy of image and protection for assistants and observers. RESULTS: The exoscope in 3D mode performed as well as or better than the microscope for image quality, field of view and manoeuvrability. It outperformed the microscope for compatibility with PPE, surgeon comfort and assistant/observer experience. It scored almost as highly as the microscope for depth perception. CONCLUSION: There is likely to be a learning curve but this initial assessment of the exoscope shows significant potential as an alternative to the operating microscope in otological surgery but with the advantage of allowing the use of appropriate PPE and better ergonomics for both surgeon and assistant/observer.

Research Techniques Made Simple: Volume Scanning Electron Microscopy.

Volume scanning electron microscopy (VSEM) involves the serial sectioning and imaging of a sample using scanning electron microscopy (SEM), followed by segmentation and three-dimensional (3D) reconstruction using computer software packages to allow visualization of 3D structures. VSEM can reveal qualitative and quantitative properties of organelles and cells within tissues at nanoscale. The ability to visualize spatial relationships of structures of interest within and across cells in 3D space in particular sets VSEM apart from conventional SEM and transmission electron microscopy. Here, we provide an overview of VSEM platforms and image processing, highlighting characteristics that will aid selection of a method to address specific research questions in dermatological research.

Label-free imaging and evaluation of characteristic properties of asthma-derived eosinophils using optical diffraction tomography.

Optical diffraction tomography (ODT), an emerging imaging technique that does not require fluorescent staining, can measure the three-dimensional distribution of the refractive index (RI) of organelles. In this study, we used ODT to characterize the pathological characteristics of human eosinophils derived from asthma patients presenting with eosinophilia. In addition to morphological information about organelles appearing in eosinophils, including the cytoplasm, nucleus, and vacuole, we succeeded in imaging specific granules and quantifying the RI values of the granules. Interestingly, ODT analysis showed that the RI (i.e., molecular density) of granules was significantly different between eosinophils from asthma patients and healthy individuals without eosinophilia, and that vacuoles were frequently found in the cells of asthma patients. Our results suggest that the physicochemical properties of eosinophils derived from patients with asthma can be quantitatively distinguished from those of healthy individuals. The method will provide insight into efficient evaluation of the characteristics of eosinophils at the organelle level for various diseases with eosinophilia.

Assessment of clinical measures of total and regional body composition from a commercial 3-dimensional optical body scanner.

BACKGROUND: The accurate assessment of total body and regional body circumferences, volumes, and compositions are critical to monitor physical activity and dietary interventions, as well as accurate disease classifications including obesity, metabolic syndrome, sarcopenia, and lymphedema. We assessed body composition and anthropometry estimates provided by a commercial 3-dimensional optical (3DO) imaging system compared to criterion measures. METHODS: Participants of the Shape Up! Adults study were recruited for similar sized stratifications by sex, age (18-40, 40-60, >60 years), BMI (under, normal, overweight, obese), and across five ethnicities (non-Hispanic [NH] Black, NH White, Hispanic, Asian, Native Hawaiian/Pacific Islander). All participants received manual anthropometry assessments, duplicate whole-body 3DO (Styku S100), and dual-energy X-ray absorptiometry (DXA) scans. 3DO estimates provided by the manufacturer for anthropometry and body composition were compared to the criterion measures using concordance correlation coefficient (CCC) and Bland-Altman analysis. Test-retest precision was assessed by root mean square error (RMSE) and coefficient of variation. RESULTS: A total of 188 (102 female) participants were included. The overall fat free mass (FFM) as measured by DXA (54.1 ± 15.2 kg) and 3DO (55.3 ± 15.0 kg) showed a small mean difference of 1.2 ± 3.4 kg (95% limits of agreement -7.0 to +5.6) and the CCC was 0.97 (95% CI: 0.96-0.98). The CCC for FM was 0.95 (95% CI: 0.94-0.97) and the mean difference of 1.3 ± 3.4 kg (95% CI: -5.5 to +8.1) reflected the difference in FFM measures. 3DO anthropometry and body composition measurements showed high test-retest precision for whole body volume (1.1 L), fat mass (0.41 kg), percent fat (0.60%), arm and leg volumes, (0.11 and 0.21 L, respectively), and waist and hip circumferences (all <0.60 cm). No group differences were observed when stratified by body mass index, sex, or race/ethnicity. CONCLUSIONS: The anthropometric and body composition estimates provided by the 3DO scanner are precise and accurate to criterion methods if offsets are considered. This method offers a rapid, broadly available, and automated method of body composition assessment regardless of body size. Further studies are recommended to examine the relationship between measurements obtained by 3DO scans and metabolic health in healthy and clinical populations.

EndoNaut two-dimensional fusion imaging with a mobile C-arm for endovascular treatment of occlusive peripheral arterial disease.

BACKGROUND: Endovascular treatment has become the first-line strategy for peripheral arterial disease (PAD). Given the number of procedures required, any technology associated with a reduction in radiation exposure and contrast volume is highly relevant. In the present study, we evaluated whether two-dimensional (2D) fusion imaging could reduce the radiation exposure and contrast volume during endovascular treatment of occlusive PAD. METHODS: Our consecutive, retrospective, single-center, nonrandomized comparative trial included patients with PAD at the femoral, popliteal, and/or tibial level, at any clinical stage, if they were candidates for endovascular revascularization. Patients were treated with or without the EndoNaut 2D fusion imaging system (Therenva, Rennes, France) in a nonhybrid room with the same Cios Alpha mobile C-arm (Siemens, Munich, Germany). The indirect dose-area product and contrast medium volume were recorded. RESULTS: Between March 2018 and April 2020, 255 patients underwent endovascular femoropopliteal revascularization with (n = 124) or without (n = 131) 2D fusion imaging. The volume of injected contrast medium (34.7 ± 13.8 mL vs 51.3 ± 26.7 mL; P < .001) and dose-area product (8.9 ± 9.9 Gy/cm2 vs 13.5 ± 14.0 Gy/cm2; P = .003) were significantly lower for the 2D fusion imaging group than for the control group. A subgroup analysis of complex (TransAtlantic Inter-Society Consensus for the Management of Peripheral Arterial Disease C/D) lesions showed similar results. Stratification of the fusion imaging group into three subgroups, according to the procedure dates, showed no effect of a potential learning curve on the operative parameters. CONCLUSIONS: The results from the present study showed a significant reduction in the contrast volume and radiation dose for endovascular treatment of PAD when applying 2D fusion imaging technology. Overall, a reduction of >30% was observed for both operative parameters, without excessive training requirements, highlighting the potential benefits of using 2D fusion imaging when performing endovascular revascularization for PAD.

Three-dimensional printing of orbital computed tomography scan images for use in ophthalmology teaching / Impressão tridimensional de imagens médicas de tomografia computadorizada para uso no ensino de oftalmologia

ABSTRACT Introduction: The use of tridimensional (3D) printing in healthcare has contributed to the development of instruments and implants. The 3D printing has also been used for teaching future professionals. In order to have a good 3D printed piece, it is necessary to have high quality images, such as the ones from Computerized Tomography (CT scan) exam, which shows the anatomy from different cuts and allows for a good image reconstruction. Purpose: To propose a protocol for creating digital files from computerized tomography images to be printed in 3D and used as didactic material in the ophthalmology field, using open-source software, InVesalius®, Blender® and Repetier-Host©. Methods: Two orbit CT scan exam images in the DICOM format were used to create the virtual file to be printed in 3D. To edit the images, the software InVesalius® (Version 3.1.1) was used to delimit and clean the structure of interest, and also to convert to STL format. The software Blender® (Version 2.80) was used to refine the image. The STL image was then sent to the Repetier-Host© (Version 2.1.3) software, which splits the image in layers and generates the instructions to print the piece in the 3D printer using the polymer polylactic acid (PLA). Results: The printed anatomical pieces printed reproduced most structures, both bone and soft structures, satisfactorily. However, there were some problems during printing, such as the loss of small bone structures, that are naturally surrounded by muscles due to the lack of support. Conclusion: Despite the difficulties faced during the production of the pieces, it was also possible to reproduce the anatomical structures adequately, which indicates that this protocol of 3D printing from medical images is viable.

RESUMO Introdução: O uso de impressão em 3-D na área da saúde tem contribuído para o desenvolvimento de instrumentos e próteses. A impressão 3-D tem sido usada para o ensino de futuros profissionais. Para se alcançar uma boa peça em 3-D, é necessário ter imagens de alta qualidade, como aquelas geradas pelo exame de Tomografia Computadorizada (TC), que mostra a anatomia sob diferentes cortes e permite uma boa reconstrução de imagem. Objetivo: Propor um protocolo para a criação de arquivos digitais a partir de imagens de tomografia computadorizada a serem impressas em 3-D e usadas como modelo de material didático oftalmológico usando software de código aberto, InVesalius®, Bender® e Repetier-Host©. Métodos: Foram utilizadas imagens em formato DICOM provenientes de dois exames de tomografia computadorizada de órbitas para a impressão tridimensional. Para manuseio das imagens, foram utilizados o InVesalius®, versão 3.1.1, para delimitar e limpar a estrutura de interesse e também para converter em formato STL. O Blender®, versão 2.80 foi usado para refinamento. A imagem em STL foi então enviada para o programa Repetier-Host, versão 2.1.3, que divide a imagem em camadas e gera as instruções para impressão da peça em ácido polilático na impressora tridimensional. Resultados: As peças anatômicas impressas reproduziram de forma satisfatória a maioria das estruturas ósseas e musculares. No entanto, houve dificuldade durante a impressão das estruturas ósseas menores, como perda de estrutura óssea pequena, que não possuíam sustentação, por serem envoltas pelo músculo. Conclusão: Apesar das dificuldades encontradas na produção dessas peças de estudo, foi possível reproduzir estruturas com fidelidade, indicando que o protocolo proposto viabiliza a impressão de imagens oriundas da tomografia computadorizada para impressão tridimensional.

Clinical Applications of Additive Manufacturing Models in Neurosurgery: a Systematic Review

Introduction Three-dimensional (3D) printing technologies provide a practical and anatomical way to reproduce precise tailored-made models of the patients and of the diseases. Those models can allow surgical planning, besides training and surgical simulation in the treatment of neurosurgical diseases. Objective The aim of the present article is to review the scenario of the development of different types of available 3D printing technologies, the processes involved in the creation of biomodels, and the application of those advances in the neurosurgical field. Methods We searched for papers that addressed the clinical application of 3D printing in neurosurgery on the PubMed, Ebsco, Web of Science, Scopus, and Science Direct databases. All papers related to the use of any additivemanufacturing technique were included in the present study. Results Studies involving 3D printing in neurosurgery are concentrated on threemain areas: (1) creation of anatomical tailored-made models for planning and training; (2) development of devices and materials for the treatment of neurosurgical diseases, and (3) biological implants for tissues engineering. Biomodels are extremely useful in several branches of neurosurgery, and their use in spinal, cerebrovascular, endovascular, neuro-oncological, neuropediatric, and functional surgeries can be highlighted. Conclusions Three-dimensional printing technologies are an exclusive way for direct replication of specific pathologies of the patient. It can identify the anatomical variation and provide a way for rapid construction of training models, allowing the medical resident and the experienced neurosurgeon to practice the surgical steps before the operation.

Dual color DMD-SIM by temperature-controlled laser wavelength matching.

Structured illumination microscopy (SIM) is a fast and gentle super-resolution fluorescence imaging technique, featuring live-cell compatible excitation light levels and high imaging speeds. To achieve SIM, spatial modulation of the fluorescence excitation light is employed. This is typically achieved by interfering coherent laser beams in the sample plane, which are often created by spatial light modulators (SLMs). Digital micromirror devices (DMDs) are a form of SLMs with certain advantages, such as high speed, low cost and wide availability, which present certain hurdles in their implementation, mainly the blazed grating effect caused by the jagged surface structure of the tilted mirrors. Recent works have studied this effect through modelling, simulations and experiments, and laid out possible implementations of multi-color SIM imaging based on DMDs. Here, we present an implementation of a dual-color DMD based SIM microscope using temperature-controlled wavelength matching. By carefully controlling the output wavelength of a diode laser by temperature, we can tune two laser wavelengths in such a way that no opto-mechanical realignment of the SIM setup is necessary when switching between both wavelengths. This reduces system complexity and increases imaging speed. With measurements on nano-bead reference samples, as well as the actin skeleton and membrane of fixed U2OS cells, we demonstrate the capabilities of the setup.

Simultaneous orientation and 3D localization microscopy with a Vortex point spread function.

Estimating the orientation and 3D position of rotationally constrained emitters with localization microscopy typically requires polarization splitting or a large engineered Point Spread Function (PSF). Here we utilize a compact modified PSF for single molecule emitter imaging to estimate simultaneously the 3D position, dipole orientation, and degree of rotational constraint from a single 2D image. We use an affordable and commonly available phase plate, normally used for STED microscopy in the excitation light path, to alter the PSF in the emission light path. This resulting Vortex PSF does not require polarization splitting and has a compact PSF size, making it easy to implement and combine with localization microscopy techniques. In addition to a vectorial PSF fitting routine we calibrate for field-dependent aberrations which enables orientation and position estimation within 30% of the Cramér-Rao bound limit over a 66 µm field of view. We demonstrate this technique on reorienting single molecules adhered to the cover slip, λ-DNA with DNA intercalators using binding-activated localization microscopy, and we reveal periodicity on intertwined structures on supercoiled DNA.

Comparison of Accuracy of Current Ten Intraoral Scanners.

There have been various developments in intraoral 3D scanning technology. This study is aimed at investigating the accuracy of 10 scanners developed from 2015 to 2020. A maxillary dental model with reference points was printed from Form 2 (FormLabs, Somerville, MA, USA). The model was scanned 5 times with each intraoral scanner (IOS); Trios 3 (normal and high-resolution mode); Trios 4 (normal and high-resolution mode) (3Shape Trios A/S, Copenhagen, Denmark); iTero Element, iTero 2, and iTero 5D Element (Align Technologies, San Jose, California, USA); Dental Wings (Dental Wings, Montreal QC, Canada); Panda 2 (Pengtum Technologies, Shanghai, China); Medit i500 (Medit Corp. Seoul, South Korea); Planmeca Emerald™ (Planmeca, Helsinki, Finland); and Aoralscan (Shining 3D Tech. Co., Ltd., Hangzhou, China). After the scan, the 3D scanned stereolithography files were created. The various distances were measured five times in X, Y, Z, and XY axes of various scans and with a vernier caliper (control) and from the Rhinoceros software. The data were analyzed using SPSS 18. Test for the normality of the various measurement data were done using Kolmogorov-Smirnov test. The trueness and precision of the measurements were compared among the various scans using the Kruskal-Wallis test. The significance was considered at P < 0.05. The trueness of the intraoral scans was analyzed by comparing the measurements from the control. Precision was tested through the measurements of repeated scans. It showed that more the distance is less the accuracy for all scanners. In all studied scanners, the trueness varied but precision was favorably similar. Diagonal scanning showed less accuracy for all the scanners. Hence, when scanning the full arch, the dentist needs to take more caution and good scan pattern. Trios series showed the best scan results compared to other scanners.

A potential method for sex estimation of human skeletons using deep learning and three-dimensional surface scanning.

Deep learning based on radiological methods has attracted considerable attention in forensic anthropology because of its superior classification capacities over human experts. However, radiological instruments are limited in their nature of high cost and immobility. Here, we integrated a deep learning algorithm and three-dimensional (3D) surface scanning technique into a portable system for pelvic sex estimation. Briefly, the images of the ventral pubis (VP), dorsal pubis (DP), and greater sciatic notch (GSN) were cropped from virtual pelvic samples reconstructed from CT scans of 1000 individuals; 80% of them were used to train and internally evaluate convolutional neural networks (CNNs) that were then evaluated externally with the remaining samples. An additional 105 real pelvises were documented virtually with a handheld 3D surface scanner, and the corresponding snapshots of the VP, DP, and GSN were predicted by the trained CNN models. The CNN models achieved excellent performance in the external testing using CT-based images, with accuracies of 98.0%, 98.5%, and 94.0% for VP, DP, and GSN, respectively. When the CT-based models were applied to 3D scanning images, they obtained satisfactory accuracies above 95% on the VP and DP images compared to the GSN with 73.3%. In a single-blind trial, a multiple design that combined the three CNN models yielded a superior accuracy of 97.1% with 3D surface scanning images over two anthropologists. Our study demonstrates the great potential of deep learning and 3D surface scanning for rapid and accurate sex estimation of skeletal remains.