Digital imaging applications in anatomic pathology

Digital imaging has progressed at a rapid rate and is likely to eventually replace chemical photography in most areas of professional and amateur digital image acquisition. In pathology, digital microscopy has implications beyond that of taking a photograph. The arguments for adopting this new medium are compelling, and given similar developments in other areas of pathology and radiologic imaging, acceptance of the digital medium should be viewed as a component of the technological evolution of the laboratory. A digital image may be stored, replicated, catalogued, employed for educational purposes, transmitted for further interpretation (telepathology), analyzed for salient features (medical vision/image analysis), or form part of a wider digital healthcare strategy. Despite advances in digital camera technology, good image acquisition still requires good microscope optics and the correct calibration of all system components, something which many neglect. The future of digital imaging in pathology is very promising and new applications in the fields of automated quantification and interpretation are likely to have profound long-term influence on the practice of anatomic pathology. This paper discusses the state of the art of digital imaging in anatomic pathology.     

How to improve microscopic images obtained with consumer-type digital cameras

AIM: Digital imaging is useful in conventional photography because it immediately provides images, and the image quality can be improved afterwards by the use of computer programs. The major disadvantages of consumer-type digital cameras mounted on microscopes are (i) unequal illumination through the image, and (ii) a coloured background. A computer program was specifically adapted and refined to improve images obtained with consumer-type digital cameras mounted on microscopes. METHODS AND RESULTS: An approach using a division operation between the specimen image and a background image leads to homogeneous illumination throughout the image, with automatically corrected brightness and white background. The correct colour spectrum is preserved by correction of the histogram. This approach was obtained from the freeware computer program 'Image Arithmetic'. In a test, three different consumer-type digital cameras (Sony, Nikon, Olympus) on different microscopes were used to obtain images of different types of histological specimens (cervical smear, bone marrow biopsy, and colonic biopsy). The computer program dramatically improved the quality of images obtained with all tested cameras. CONCLUSION: Using this approach, even low-cost digital cameras mounted on microscopes produce brilliant images with homogeneous illumination and a white background, the image quality being comparable with expensive cameras especially designed for microscopes.     

Digital pathology: current status and future perspectives

During the last decade pathology has benefited from the rapid progress of image digitizing technology. The improvement in this technology had led to the creation of slide scanners which are able to produce whole slide images (WSI) which can be explored by image viewers in a way comparable to the conventional microscope. The file size of the WSI ranges from a few megabytes to several gigabytes, leading to challenges in the area of image storage and management when they will be used routinely in daily clinical practice. Digital slides are used in pathology for education, diagnostic purposes (clinicopathological meetings, consultations, revisions, slide panels and, increasingly, for upfront clinical diagnostics) and archiving. As an alternative to conventional slides, WSI are generally well accepted, especially in education, where they are available to a large number of students with the full possibilities of annotations without the problem of variation between serial sections. Image processing techniques can also be applied to WSI, providing pathologists with tools assisting in the diagnosis-making process. This paper will highlight the current status of digital pathology applications and its impact on the field of pathology.     

Telemedicine screening of diabetic retinopathy using a hand-held fundus camera.

The objective was to evaluate digital images of the retina from a handheld fundus camera (Nidek NM-100) for suitability in telemedicine screening of diabetic retinopathy. A handheld fundus camera (Nidek) and a standard fundus camera (Zeiss) were used to photograph 49 eyes from 25 consecutive patients attending our diabetic clinic. One patient had cataracts, making it impossible to get a quality image of one of the eyes (retina). The Nidek images were digitized, compressed, and stored in a Fujix DF-10M digitizer supplied with the camera. The digital images and the photographs were presented separately in a random order to three ophthalmologists. The quality of the images was ranked as good, acceptable or unacceptable for diabetic retinopathy diagnosis. The images were also evaluated for the presence of microaneurysms, blot hemorrhages, exudates, fibrous tissue, previous photocoagulation, and new vessel formation. kappa Values were computed for agreement between the photographs and digital images. Overall agreement between the photographs and digital images was poor (kappa < 0.30). On average, only 24% of the digital images were graded as being good quality and 56% as having an acceptable quality. However, 93% of the photographs were graded as good-quality images for diagnosis. The results indicate that the digital images from the handheld fundus camera may not be suitable for diagnosis of diabetic retinopathy. The images shown on the liquid crystal display (LCD) screen of the camera were of good quality. However, the images produced by the digitizer (Fujix DF-10M) attached to the camera were not as good as the images shown on the LCD screen. A better digitizing system may produce better quality images from the Nidek camera.     

Technical Challenges of Enterprise Imaging: HIMSS-SIIM Collaborative White Paper

This white paper explores the technical challenges and solutions for acquiring (capturing) and managing enterprise images, particularly those involving visible light applications. The types of acquisition devices used for various general-purpose photography and specialized applications including dermatology, endoscopy, and anatomic pathology are reviewed. The formats and standards used, and the associated metadata requirements and communication protocols for transfer and workflow are considered. Particular emphasis is placed on the importance of metadata capture in both order- and encounter-based workflow. The benefits of using DICOM to provide a standard means of recording and accessing both metadata and image and video data are considered, as is the role of IHE and FHIR.     

Use of virtual microscopy for didactic live-audience presentation in anatomic pathology

Didactic presentations on the topic of anatomic pathology in front of a live audience have been largely dependent on the use of standard 2 x 2 inch projection slides (Kodachromes) of selected still images from the topic at hand. Because of the highly visual nature of the specialty of anatomic pathology, this method has had some serious limitations. With the advent of digital imaging techniques and the availability of new electronic software for the projection of images, new possibilities have become available for didactic presentations in anatomic pathology in front of a large, live audience. We describe a method whereby large digital images or "virtual slides" were produced from digitally scanned whole-mount sections of histologic glass slides and projected using a combination of PowerPoint (Microsoft Corp, Redmond, WA) and virtual microscopy in front of a live audience. To provide a seamless transition between the two presentation formats, the personal computer-based PowerPoint slides were hyperlinked to a browser-based virtual microscope viewer. The presenter, with the use of a mouse, was able to "move" the image of the scanned slide on the screen, to transition seamlessly among various magnifications, and to rapidly select from the whole-mount scanned slide among any areas of interest pertinent to the topic. Thus, the visual experience obtained by the audience simulated that of viewing a glass slide at a multi-headed microscope during a glass slide tutorial. Because this most closely approximates the experience of reviewing glass slides under the microscope for practicing pathologists, the educational experience of the presentation is greatly enhanced by the use of this technique. Also, this method permits making this type of presentation available to a much larger group of individuals in a live audience.     

The use of digital images in pathology

Digital images are routinely used by the publishing industry, but most diagnostic pathologists are unfamiliar with the technology and its possibilities. This review aims to explain the basic principles of digital image acquisition, storage, manipulation and use, and the possibilities provided not only in research, but also in teaching and in routine diagnostic pathology. Images of natural objects are usually expressed digitally as ‘bitmaps’—rectilinear arrays of small dots. The size of each dot can vary, but so can its information content in terms, for example, of colour, greyscale or opacity. Various file formats and compression algorithms are available. Video cameras connected to microscopes are familiar to most pathologists; video images can be converted directly to a digital form by a suitably equipped computer. Digital cameras and scanners are alternative acquisition tools of relevance to pathologists. Once acquired, a digital image can easily be subjected to the digital equivalent of any conventional darkroom manipulation and modern software allows much more flexibility, to such an extent that a new tool for scientific fraud has been created. For research, image enhancement and analysis is an increasingly powerful and affordable tool. Morphometric measurements are, after many predictions, at last beginning to be part of the toolkit of the diagnostic pathologist. In teaching, the potential to create dramatic yet informative presentations is demonstrated daily by the publishing industry; such methods are readily applicable to the classroom. The combination of digital images and the Internet raises many possibilities; for example, instead of seeking one expert diagnostic opinion, one could simultaneously seek the opinion of many, all around the globe. It is inevitable that in the coming years the use of digital images will spread from the laboratory to the medical curriculum and to the whole of diagnostic pathology. © 1997 John Wiley & Sons, Ltd.     

Going digital: Image preparation for biomedical publishing

Authors are more often being held responsible for readying their own data figures for digital publication by scanning them at the proper resolution and preparing them for presentation in both print and on‐line journals. In this manner, the visuals can be printed at the highest quality the publisher can provide and be ready for rapid electronic distribution on the Internet. Therefore, authors must become knowledgeable in the visual preparation process in order to generate electronic images that will be as true a representation of the original image as possible. Perfecting this procedure can be a learning experience and often requires some experimentation. When accomplished, the author will have more control of exactly how the images will look before they are published. In addition to the scan resolution, the type of digital scanner and software applications used are very important, and instruction manuals should be followed closely so as to understand the full potential of the digitizing equipment. Anat Rec (New Anat): 257:128–136, 1999. © 1999 Wiley‐Liss, Inc.     

Going digital: image preparation for biomedical publishing.

Authors are more often being held responsible for readying their own data figures for digital publication by scanning them at the proper resolution and preparing them for presentation in both print and on-line journals. In this manner, the visuals can be printed at the highest quality the publisher can provide and be ready for rapid electronic distribution on the Internet. Therefore, authors must become knowledgeable in the visual preparation process in order to generate electronic images that will be as true a representation of the original image as possible. Perfecting this procedure can be a learning experience and often requires some experimentation. When accomplished, the author will have more control of exactly how the images will look before they are published. In addition to the scan resolution, the type of digital scanner and software applications used are very important, and instruction manuals should be followed closely so as to understand the full potential of the digitizing equipment. Anat Rec (New Anat): 257:128-136, 1999.     

A time-delay integration charge-coupled device camera for slot-scanned digital radiography

We have developed a low-noise digital camera based on a 512 x 96 element CCD operating in the time-delay integration mode. This camera has been combined with an x-ray image intensifier to record radiographic images produced by a scanning slot beam of radiation. This results in the rejection of a large fraction of scattered radiation, without a significant increase in x-ray tube heat loading or image acquisition time. Here we describe the design of our CCD camera and the results of our investigations of camera resolution, linearity, noise, and quantum efficiency. We have found that both the resolution limit (50 mm-1) and the dynamic range (2100) of this novel camera are greater than reported values for conventional video cameras. Applications of this system in digital angiography and mammography are discussed.     

Use of a digital film scanner to enhance low-power bright field photomicrography

 Low-power bright field photomicrographs often suffer from insufficient sharpness, uneven illumination, and colour hues. Using a film scanner, commercially available and designed for digitizing 35-mm transparencies, we directly scanned microscopic slides that carried dye-labelled and stained sections. The digital images covered a field of up to 24×36 mm and revealed excellent sharpness, absolutely even illumination and superior colour reproduction as compared to conventional photomicrographs taken with binoculars, macro lenses, or microscopes. As the method requires neither specialized instrumentation nor expert knowledge of photomicrographic techniques, it reduces costs and saves time. The high-quality digital survey micrographs can easily be used for image processing, image analysis and morphometry. Thus, this new method is valuable not only for pathology, embryology, histochemistry, and the neurosciences, but also for the exchange of low-power micrographs via the internet and for computer media that are increasingly used in medical education. Accepted: 17 April 1998     

Virtual microscopy: high resolution digital photomicrography as a tool for light microscopy simulation

Recent advances in microcomputers and high resolution digital video cameras provide pathologists the opportunity to combine precision optics with digital imaging technology and develop new educational and research tools. We review recent advances in virtual microscopy and describe techniques for viewing digital images using a microcomputer-based workstation to simulate light microscopic examination, including scanning at low power to select features of interest and zooming to increase magnification. Hardware and software components necessary to acquire digital images of histological and cytological slides, and closely simulate their examination under a light microscope are discussed. The workstation is composed of a MicroLumina digital scanning camera (Leaf Systems, Southborough, MA), light microscope (Olympus Optical Co., Lake Success, NY), Pentium (Intel Corp., Santa Clara, CA) 166 MHz microcomputer configured with 64 megabytes of random access memory (RAM), a MGA Millenium Powerdesk graphics card (Matrox Graphics, Inc., Montreal, Canada) and Photoshop software (Adobe Systems Inc., San Jose, CA) running in a Windows 95 (Microsoft Corp., Redmond, WA) environment. Images with spatial resolutions of up to 2700 x 3400 pixels in 36-bit color, can be displayed simultaneously as distinct images in a montage, or merged into a single composite image file to highlight significant features of a histological or cytological slide. These image files are saved in Joint Photographers Experts Group (JPEG) format using compression ratios of up to 80:1 without detectable visual degradation. The advantages and technical limitations of various workstation components are addressed and applications of this technology for pathology education, proficiency testing, telepathology, and database development are discussed.     

System for digital acquisition of gastrointestinal images

Previous theoretical work and clinical experience with digital acquisition of fluoroscopic images have identified several problems which needed to be solved. These are: image resolution; blurring due to patient motion, combined with long exposure times; and excessive x-ray quantum mottle levels. We will show that application of pulsed progressive readout (PPR) methods to the TV camera solves these problems. By permitting a high-intensity x-ray pulse to be delivered, all motion is stopped and quantum mottle is reduced to acceptable levels. It will be shown that 1024 x 1024 digital matrices provide adequate resolution and 8-bit digitization is sufficient to permit the same quality as is used in conventional 100-mm photofluorography. User acceptance can be made easier by incorporation of existing photofluorographic controls (with which the radiologist is already familiar) to acquire the digital images. It is possible to interface PPR video systems using existing 100-mm exposure circuits without much modification and the resulting system can be regarded as a digital 100-mm camera.     

Semi-automatic medical image processing

During the last few years a number of systems for graphics generation and image processing have been developed by the authors. Depending on the application, one can consider many kinds of systems; for example for simple analysis of cellular images, a system that realizes the video mixing between a camera and graphics data from a microcomputer would be adequate. For a more elaborate analysis, such as angiographic and echographic images requiring a high level of interactivity, a semi-automatic acquisition system linked to a host computer would be necessary. Limits in precision and execution time of these semi-automatic systems leads to independent work-stations that realize digital acquisition, processing, and display of images.     

The Right Mediastinal Border and Central Venous Anatomy on Frontal Chest Radiograph—Direct CT Correlation

We describe a direct and accurate method for defining chest radiographic anatomy and use this method to delineate the anatomic composition of the right mediastinal border in an adult population. Intravenous contrast-enhanced computed tomographic scans of the chest and accompanying scout tomograms from 99 adults without previously known or detected cardiopulmonary disease that could potentially distort mediastinal, cardiac, or pulmonary anatomy were retrospectively evaluated. Transverse CT images through the mediastinum were directly referenced to the respective acquisition location on the scout tomogram via the acquisition reference line. The anatomic composition of the right mediastinal border on the scout tomogram was determined by drawing a vertical line tangential to the most lateral right mediastinal structure in each transverse CT image. The lengths and relationships of these structures were tabulated. These results will help to create a consensus among radiologists and other clinicians regarding radiographic anatomy, allowing improved localization of mediastinal pathology and enabling more optimal positioning of vascular and cardiac support devices.     

Image quality in digital chromosome analysis systems

This paper reports on an investigation into the differences in image quality of different components used in a digital image processing system for chromosome analysis. As chromosome aberrations are important tools in the cloning of genes, it is important to know if the introduction of computerized analysis systems increases the risk of missing small aberrations. In this investigation the number of visible bands on a number of chromosomes has been used as a measure of quality. The images compared are microscope ocular images, photographs from a microscope builtin camera, digital images from a high and from a standard resolution camera, presented both on screen and print-out on paper. The main conclusions are that: (1) the view in the microscope ocular gives the best resolution, (2) there are risks of losing vital information using the digital image processing system for chromosome analysis, and (3) this risk is significantly reduced when using a high resolution camera.     

构建高分辨率中国数字人男性数据集

目的构建高分辨率中国男性数字人0.2mm断层间距图像数据集。方法取捐献健康男性遗体,经颈动脉红色灌注、低温冷冻定型、专用包埋模具垂直包埋、一次装夹连续等间距铣削、PhaseOneH25数码相机采集连续断层数据以构建数据集。结果共获得9320个RAW格式断层图像数据,分辨率为4080×5440象素,RAW格式单层数据量为27.5M,RAW格式总数据量为260G。结论高分辨率中国数字人男性数据集由于采用了2200万象素的高分辩率数码相机,断层上如血管、脂肪、软骨和骨骼等组织的边界清晰。并可用随机软件对图像的色彩、曝光量以及断层感兴趣区域进行重新处理。     

Displaying radiologic images on personal computers: Practical applications and uses

This is the fifth and final article in our series for radiologists and imaging scientists on displaying, manipulating, and analyzing radiologic images on personal computers (PCs). There are many methods of transferring radiologic images into a PC, including transfer over a network, transfer from an imaging modality storage archive, using a frame grabber in the image display console, and digitizing a radiograph or 35-mm slide. Depending on the transfer method, the image file may be an extended gray-scale contrast, 16-bit raster file or an 8-bit PC graphics file. On the PC, the image can be viewed, analyzed, enhanced, and annotated. Some specific uses and applications include making 35-mm slides, printing images for publication, making posters and handouts, facsimile (fax) transmission to referring clinicians, converting radiologic images into medical illustrations, creating a digital teaching file, and using a network to disseminate teaching material. We are distributing a 16-bit image display and analysis program for Macintosh computers, Dr Razz, taht illustrates many of the principles discussed in this review series. The program is available for no charge by anonymous file transfer protocol (ftp).     

Acquiring images with very high resolution using a composing method

Composing microscopic images of very high resolution from several parts posed some problems. One of them was the necessity to adjust the focusing level when moving from one part to another. Re-focusing lead to problems with joining the image parts, which did not correspond exactly, and the area of image fusion was noticeable. A computer program was developed to overcome these problems. Our program worked with all the image parts together to find their optimal order for image fusion. Individual image parts were joined using a steep gradient running along a randomly generated curve. This method gave good results even in images with background or holes in the tissue. The method of composing large images from individual parts was used for digitizing the skin lymphoma collection of the Institute of Dermatology, University Hospital, Zürich. This collection of digital images is a part of the 6th version of Hypertext atlas of Dermatopathology at www.muni.cz/atlases.     

The x-ray light valve: a low-cost digital radiographic imaging system

In recent years new digital x-ray radiographic and fluoroscopic systems based on large-area flat-panel technology have revolutionized our capability of producing x-ray images. However, such imagers are extraordinarily expensive and their rapid image acquisition capability is not required for many applications such as radiography. Here we report a novel approach to achieve a high-quality digital radiographic system at a cost which is only a small fraction of competitive digital technologies. The results demonstrate that our proposed x-ray light valve system has excellent spatial resolution and adequate sensitivity compared to existing technologies.