Development of a 3D optical scanner for evaluating patient-specific dose distributions

PurposeThis paper describes the hardware and software characteristics of a 3D optical scanner (P3DS) developed in-house. The P3DS consists of an LED light source, diffuse screen, step motor, CCD camera, and scanner management software with 3D reconstructed software.Materials and methodWe performed optical simulation, 2D and 3D reconstruction image testing, and pre-clinical testing for the P3DS. We developed the optical scanner with three key characteristics in mind. First, we developed a continuous scanning method to expand possible clinical applications. Second, we manufactured a collimator to improve image quality by reducing scattering from the light source. Third, we developed an optical scanner with changeable camera positioning to enable acquisition of optimal images according to the size of the gel dosimeter.ResultsWe confirmed ray-tracing in P3DS with optic simulation and found that 2D projection and 3D reconstructed images were qualitatively similar to the phantom images. For pre-clinical tests, the dose distribution and profile showed good agreement among RTP, optical CT, and external beam radiotherapy film data for the axial and coronal views. The P3DS has shown that it can scan and reconstruct for evaluation of the gel dosimeter within 1 min. We confirmed that the P3DS system is a useful tool for the measurement of 3D dose distributions for 3D radiation therapy QA. Further experiments are needed to investigate quantitative analysis for 3D dose distribution.     

Geographic and gender shape differences in the carapace of Liocarcinus depurator (Brachyura: Portunidae) using geometric morphometrics and the influence of a digitizing method

The current work describes the use of geometric morphometrics in the study of the portunid crab Liocarcinus depurator carapace shape variation along the Mediterranean coast of Spain. As a first step, the differences in shape induced by digitizing the carapace with either a digital camera or a flat bed scanner were investigated. Carapace shape information inferred from the camera images proved different from that obtained from the scanner images. This difference was greater than any shape variation because of crab gender or sample location. Carapace shape was analysed using multiple analysis of covariance (with centroid size as a covariate) of the partial warps and uniform components indicating differences between genders and between sample location; however, significant three-way interaction implied that such differences were gender and location specific. Landmarks identifying carapace width, carapace length and posterior carapace width showed greatest variation; hence gender and location differences were further analysed through the interrelationships between these linear measures. Regression analysis of the relationship between posterior carapace width and the ratio of carapace width to carapace length indicated clear differences between locations but not between genders. Crabs from the Alborán Sea had proportionally wider posterior carapaces than those from Alicante or North Catalonia. This finding may represent the difference between a southern influx of an Atlantic population distinguishable from a more northerly Mediterranean one, which would be in agreement with the overall oceanographic surface circulation pattern in the western Mediterranean.     

High performance CCD camera system for digitalisation of 2D DIGE gels

An essential step in 2D DIGE‐based analysis of differential proteome profiles is the accurate and sensitive digitalisation of 2D DIGE gels. The performance progress of commercially available charge‐coupled device (CCD) camera‐based systems combined with light emitting diodes (LED) opens up a new possibility for this type of digitalisation. Here, we assessed the performance of a CCD camera system (Intas Advanced 2D Imager) as alternative to a traditionally employed, high‐end laser scanner system (Typhoon 9400) for digitalisation of differential protein profiles from three different environmental bacteria. Overall, the performance of the CCD camera system was comparable to the laser scanner, as evident from very similar protein abundance changes (irrespective of spot position and volume), as well as from linear range and limit of detection.     

Evaluation of a CCTV device for partial sight

Closed circuit TV has proved to be a valuable aid for many patients with partial sight and a new device for magnification of printed material has been developed. This is in the form of a small camera, guided over the page by the subject's hand and incorporating a roller drive for a scanning mechanism. The scanning head fits into the hand easily; it is linked to an electronic TV receiver in the form of a mosaic. Tracking the scanner manually operates a flow of the display to the left, with new material emerging from the right of the screen. The present paper outlines an evaluation of the device under different conditions with patients.     

Guide du bon usage de la TDM en médecine nucléaire. IntroductionCT good practices for hybrid nuclear medicine (SPECT/CT and PET/CT): Introduction

Nuclear medicine hybrid imaging is a technological evolution of gamma camera scintigraphy or positron emission tomography imaging methods that are now often coupled with an anatomical imaging device, essentially a CT scanner. Following a large demand from the nuclear physicians themselves, but also from the French Nuclear Safety Authority, this guide is intended for the entire nuclear medicine community to integrate both the aspects of radiation protection related to coupled CT and those related to the quality of the CT images according to the clinical context.     

A cost-effective maize ear phenotyping platform enables rapid categorization and quantification of kernels

High-throughput phenotyping systems are powerful, dramatically changing our ability to document, measure, and detect biological phenomena. Here, we describe a cost-effective combination of a custom-built imaging platform and deep-learning-based computer vision pipeline. A minimal version of the maize (Zea mays) ear scanner was built with low-cost and readily available parts. The scanner rotates a maize ear while a digital camera captures a video of the surface of the ear, which is then digitally flattened into a two-dimensional projection. Segregating GFP and anthocyanin kernel phenotypes are clearly distinguishable in ear projections and can be manually annotated and analyzed using image analysis software. Increased throughput was attained by designing and implementing an automated kernel counting system using transfer learning and a deep learning object detection model. The computer vision model was able to rapidly assess over 390 000 kernels, identifying male-specific transmission defects across a wide range of GFP-marked mutant alleles. This includes a previously undescribed defect putatively associated with mutation of Zm00001d002824, a gene predicted to encode a vacuolar processing enzyme. Thus, by using this system, the quantification of transmission data and other ear and kernel phenotypes can be accelerated and scaled to generate large datasets for robust analyses.     

Quantifying fungal infection of plant leaves by digital image analysis using Scion Image software

A digital image analysis method previously used to evaluate leaf color changes due to nutritional changes was modified to measure the severity of several foliar fungal diseases. Images captured with a flatbed scanner or digital camera were analyzed with a freely available software package, Scion Image, to measure changes in leaf color caused by fungal sporulation or tissue damage. High correlations were observed between the percent diseased leaf area estimated by Scion Image analysis and the percent diseased leaf area from leaf drawings. These drawings of various foliar diseases came from a disease key previously developed to aid in visual estimation of disease severity. For leaves of Nicotiana benthamiana inoculated with different spore concentrations of the anthracnose fungus Colletotrichum destructivum, a high correlation was found between the percent diseased tissue measured by Scion Image analysis and the number of leaf spots. The method was adapted to quantify percent diseased leaf area ranging from 0 to 90% for anthracnose of lily-of-the-valley, apple scab, powdery mildew of phlox and rust of golden rod. In some cases, the brightness and contrast of the images were adjusted and other modifications were made, but these were standardized for each disease. Detached leaves were used with the flatbed scanner, but a method using attached leaves with a digital camera was also developed to make serial measurements of individual leaves to quantify symptom progression. This was successfully applied to monitor anthracnose on N. benthamiana leaves. Digital image analysis using Scion Image software is a useful tool for quantifying a wide variety of fungal interactions with plant leaves.     

Review of different platforms to perform rapid onsite evaluation via telecytology

There is increased utilisation of cytopathology to provide a rapid onsite evaluation (ROSE) of fine needle aspiration and touch preparations of small biopsies. A well‐executed ROSE procedure can significantly impact the diagnostic quality and appropriate specimen triage of procured biopsy materials. To accommodate the demand for ROSE, telecytology has been increasingly implemented to facilitate ROSE occurring remotely. Telecytology can be categorised based on camera systems including eyepiece system, camera port system and robotic microscope/whole slide image scanner system. Image sharing methods include static images, broadcast only live video streaming, teleconferencing and whole slide image management system. In this review, we will discuss the advantages and disadvantages of each of these systems and deployment considerations.     

A television/computer three-dimensional surface shape measurement system

An optical scanner is described which has been designed primarily for the measurement of human back shape. A projector and television camera were mounted together in a box which could rotate about a horizontal axis. The projector shone a horizontal plane of light, which was viewed at an angle from below by the television camera, linked directly to a minicomputer. The shape of the line of light formed by the plane as it fell on an object, together with a knowledge of the geometry of the system, enabled three-dimensional coordinates of points on the line to be calculated. A record of a surface shape was built up by scanning the object in about 2 s. Calibration of the system was achieved by scanning an object of known dimensions. Sets of algorithms are described which derive geometric parameters from the calibration scan and which sort surface shape coordinates, outline them and detect special markers from the surface shape scan. The accuracy of measurement exceeded the design aim of +/- 3 mm in each axis within a volume of 400 mm x 500 mm x 300 mm.     

Specialized software product for comparative analysis of multicomponent DNA fingerprints

GelAnalyzer software, which is used to identify and correctly compare DNA fingerprints consisting of a large number of discrete bands, has been developed by the project to study the quantitative changes in DNA polymorphism patterns in animals and humans exposed to gamma radiation. The actual capabilities of this program are much broader and include the possibility to analyze the images of any multicomponent gels containing fragments of DNA, RNA, and proteins. This software product runs on Windows. GelAnalyzer allows one to analyze gel images obtained by a scanner, camera, or digital camera and ensures the visual control of the identification and comparative analysis of bands; it also makes it possible to take into account the bands that are poorly identified automatically and exclude the artifacts (incidental marks) on images. The operation of GelAnalyzer software is based on the determination of the values of normalized coordinates of bands with allowance for the relative electrophoretic mobility (Rf) of PCR products and comparison of their spectra (set of bands in gel lanes) to reveal the similarities or differences in their components with subsequent statistical data processing and display the results of the analysis.     

Imaging technologies for the detection of multiple stains in proteomics

Laser-based scanners and charge-coupled device (CCD) camera systems are evolving to have greater functional capabilities for capturing images from a range of staining technologies used in gel electrophoresis and electroblotting. Digitizing Coomassie Brilliant Blue (CBB) stained gels and silver stained gels has now become possible using a laser-based gel scanner, the FLA-5000 fluorescent image analyzer system. Also, a simultaneous dual fluorescent imaging function has been incorporated into the FLA-5000 system, utilizing dichroic mirrors with both the optical system and the emission filter. In the workflow of routine proteomics research, the relationship between SYPRO dye staining and fluorescent detection using the FLA-5000 system have become symbiotic. Additionally in many cases, subsequent staining of the gel with CBB is useful for future research, and thus imaging instruments should be able to handle both staining formats. Digitizing the CBB stained gel can now be easily performed by the FLA-5000 fluorescent image analyzer system using a fluorescent board as an epi-illumination background. A cooled CCD camera system has the potential of imaging not only chemiluminescent membranes but also digitizing molecular weight markers and fluorescent detection of SYPRO dye-stained gels. With Multi Gauge software version 2.0 it is now a simple task to combine two images into one, as commonly required in dual detection experiments. The LAS-3000 system was designed to capture chemiluminescent images and to digitize the images automatically. Thus, new capabilities added to gel imaging systems make them capable of detecting and displaying multiple signals more conveniently.     

Profiling Expression Patterns and Isolating Differentially Expressed Genes by cDNA Microarray System with Colorimetry Detection

A high-density cDNA microarray with colorimetry detection system to simultaneously monitor the expression of many genes on nylon membrane is described and characterized. To quantify the expression of genes and to isolate differentially expressed genes, the southern hybridization process on filter membranes was employed. The levels of gene expression were represented by color intensities generated by colorimetric reactions in place of hazardous radioisotopes or costly laser-induced fluorescence detection. The gene expression patterns on nylon membranes were digitized by devices such as an economical flatbed scanner or a digital camera. The quantitative information of gene expression was retrieved by image analysis software. Quantitative comparison of the northern dot-blotting method with the microarray system is described. Applications employing single-color detection as well as dual-color detection to isolate differentially expressed genes among thousands of genes are demonstrated.     

Gallium-67 Scans in the Diagnosis of Pyelonephritis

Gallium-67 (⁶⁷Ga) citrate was administered intravenously (50 microcuries per kg of body weight) to patients in whom acute and chronic urinary tract infections were suspected. Scanning was done, using both the Anger-type scintillation camera and the rectilinear scanner, 24 to 78 hours after injection of the isotope.The preliminary results imply that ⁶⁷Ga renal uptake is present in patients with pyelonephritis whether overt or silent, as well as in patients with uretero-sigmoidostomies. However, ⁶⁷Ga renal uptake is not present in patients with radiographic evidence of chronic pyelonephritis without active infection and in patients without renal disease.     

Automated image acquisition and processing using a new generation of 4K x 4K CCD cameras for cryo electron microscopic studies of macromolecular assemblies

We have previously reported the development of AutoEM, a software package for semi-automated acquisition of data from a transmission electron microscope. In continuing efforts to improve the speed of structure determination of macromolecular assemblies by electron microscopy, we report here on the performance of a new generation of 4 K CCD cameras for use in cryo electron microscopic applications. We demonstrate that at 120 kV, and at a nominal magnification of 67000 x, power spectra and signal-to-noise ratios for the new 4 K CCD camera are comparable to values obtained for film images scanned using a Zeiss scanner to resolutions as high as approximately 1/6.5A(-1). The specimen area imaged for each exposure on the 4 K CCD is about one-third of the area that can be recorded with a similar exposure on film. The CCD camera also serves the purpose of recording images at low magnification from the center of the hole to measure the thickness of vitrified ice in the hole. The performance of the camera is satisfactory under the low-dose conditions used in cryo electron microscopy, as demonstrated here by the determination of a three-dimensional map at 15 A for the catalytic core of the 1.8 MDa Bacillus stearothermophilus icosahedral pyruvate dehydrogenase complex, and its comparison with the previously reported atomic model for this complex obtained by X-ray crystallography.     

A novel validation and calibration method for motion capture systems based on micro-triangulation

Motion capture systems are widely used to measure human kinematics. Nevertheless, users must consider system errors when evaluating their results. Most validation techniques for these systems are based on relative distance and displacement measurements. In contrast, our study aimed to analyse the absolute volume accuracy of optical motion capture systems by means of engineering surveying reference measurement of the marker coordinates (uncertainty: 0.75 mm). The method is exemplified on an 18 camera OptiTrack Flex13 motion capture system. The absolute accuracy was defined by the root mean square error (RMSE) between the coordinates measured by the camera system and by engineering surveying (micro-triangulation). The original RMSE of 1.82 mm due to scaling error was managed to be reduced to 0.77 mm while the correlation of errors to their distance from the origin reduced from 0.855 to 0.209. A simply feasible but less accurate absolute accuracy compensation method using tape measure on large distances was also tested, which resulted in similar scaling compensation compared to the surveying method or direct wand size compensation by a high precision 3D scanner. The presented validation methods can be less precise in some respects as compared to previous techniques, but they address an error type, which has not been and cannot be studied with the previous validation methods.     

Software development of the EMI head scanner.

A method has been developed to run the general purpose operating system RDOS on the same disc of the head scanner computer as is used for scanner software and data. This made it possible to develop additional software in high level programming language for image processing, based on original image data on the disc. All new images produced by the program are stored on the disc in the same format as the original images. This makes it possible to handle processed images exactly as the original ones and to do multiple operations. The following processing has been included in the program so far: subtraction, smoothing, density profiles, vertical reconstructions, magnification and labelling. A set of operator commands has been developed which are very similar to the ordinary commands for the scanner, which makes the program to appear being a direct extension of the standard scanner software.     

Quantification of nucleic acids on nitrocellulose membranes with time-resolved fluorometry.

We use a streptavidin-based macromolecular complex (SBMC) labelled with the europium chelate of 4,7-bis (chlorosulfophenyl)-1,10-phenanthroline-2,9-dicarboxylic acid (BCPDA) as a staining reagent for biotinylated DNA present on nitrocellulose filters. The fluorescent spots or bands obtained can either be observed under UV illumination, photographed by instant camera photography or quantified by using a specially designed instrument working as a high resolution time-resolved fluorometric scanner. The detection limit is approximately 10 pg of target DNA. Various experiments with use of biotinylated DNA probes hybridized to Southern transferred targets have shown that the new procedure is a useful versatile non-isotopic methodology for staining DNA on solid supports.     

Photogrammetry for 3D digitizing bones of mounted skeletons: Potential and limits

Since the late 20th century, new technologies have provided powerful ways to digitize biological structures in three dimensions (3D). Among those, photogrammetry is a low cost and non-destructive method, which has become increasingly used since the development of the digital camera. Recent studies have demonstrated that reconstructions of isolated elements can be of as high quality as those obtained with laser scanners. Here, we wanted to test the performance of photogrammetry for the quantitative analysis of mounted specimens in museum exhibitions. Indeed, access to material can be an issue in comparative anatomy and, especially, in paleontology. This is notably the case for large, impressive specimens. We performed reconstructions based on acquisitions done under various conditions and also tested the reconstruction performance of two software programs. The resulting 3D models were then compared to a reference object corresponding to the bone of interest digitized with a cutting-edge surface scanner. Our results show that photogrammetry enables quality reconstruction of the almost entire surface of the mounted bone of interest. Photogrammetry thus appears a reliable method perfectly suited to study large specimens exposed in museum gallery.     

Testing different 3D techniques using geometric morphometrics: Implications for cranial fluctuating asymmetry in humans

This study aimed to test the performance of 3D digitizer, CT scanner, and surface scanner in detecting cranial fluctuating asymmetry. Sets of 32 landmarks (6 in the midline and 13 bilateral) were acquired from 14 archeological crania using a 3D digitizer, and from 3D models generated from a CT scanner and surface scanner using Viewbox 4. Levels of shape variation were analyzed in MorphoJ using Procrustes analysis of variance and Principal component analysis. Intra-observer error accounted for 1.7%, 1.8%, and 4.5% of total shape variation for 3D digitizer, CT scanner, and surface scanner respectively. Fluctuating asymmetry accounted for 15%–16% of total shape variation. Variation between techniques accounted for 18% of total shape variation. We found a higher level of missing landmarks in our surface scan data than for both 3D digitizer and CT scanner data, and both 3D model-based techniques sometimes obscured taphonomic damage. All three 3D techniques are appropriate for measuring cranial fluctuating asymmetry. We advise against combining data collected with different techniques.     

Making blind robots see: the synergy between fluorescent dyes and imaging devices in automated proteomics

Proteomics investigations endeavor to provide a global understanding of gene product synthesis rate, degradation rate, functional competence, posttranslational modification, subcellular distribution and physical interactions with other cell components. Protein expression encompasses an enormous dynamic range. Since rare proteins cannot be amplified by any type of PCR method, sensitive detection is critical to proteome projects. Fluorescence methods deliver streamlined detection protocols, superior detection sensitivity, broad linear dynamic range and excellent compatibility with modern microchemical identification methods such as mass spectrometry. Two general approaches to fluorescence detection of proteins are currently practiced: the covalent derivatization of proteins with fluorophores or noncovalent interaction of fluorophores either via the SDS micelle or through direct electrostatic interaction with proteins. One approach for quantifying fluorescence is to use a photomultiplier tube detector combined with a laser light scanner. In addition, fluorescence imaging is performed using a charge-coupled device camera combined with a UV light or xenon arc source. Fluorescent dyes with bimodal excitation spectra may be broadly implemented on a wide range of analytical imaging devices, permitting their widespread application to proteomics studies and incorporation into semiautomated analysis environments.