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Microscopic Image Photography Techniques of the Past, Present, and Future.

The field of pathology is driven by images, both in gross pathology and in microscopic findings. Trainees learn from exposure to a variety of pathologic entities through their clinical duties, in textbooks, and from online tutorials, lectures, and interdepartmental conferences. Practicing pathologists use publications and national or international conferences to stay abreast of new developments in the field, where methods of sharing pathology information often rely heavily on microscopic images to illustrate key points. Throughout the history of pathology, photographic representation of microscopic findings has been a central focus in education, communication, and practice.


In the early 17th century, a device called the Magic Lantern, or Sciopticon, used an external light source to project semitransparent images painted on glass panes onto walls or screens. In the 1840s, the Philadelphian daguerreotypist brothers William and Frederick Langenheim adapted a method invented by Frenchman Niepce de St. Victor of making a transparent negative on glass, which was then used to print a positive image onto a second pane of glass, thus creating a transparent black and white image suitable for projection with the Magic Lantern projector. The brothers patented the method they dubbed hyalotyping (hyalo: Greek for glass) in 1850. The use of hyalotypes with the Magic Lantern projector resulted in the informal, yet better-known name lantern slides. Early lantern slides were made using albumen to coat the glass plates; a wet-collodion coating was developed shortly thereafter, followed by a dry colloid that enabled lantern slide kits to be mass produced for amateur use. Although the Langenheims envisioned using hyalotypes for entertainment, they were quickly adopted for educational and scientific purposes. (1)

The Edinger Apparatus (Figure 1, A) is the most notable device used to adapt a lantern slide camera to the microscope. Designed by Professor Ludwig Edinger of the Neurologic Institute in Frankfurt, Germany, it was advertized for use in "projecting microscopic objects, projecting lantern slides, taking photo-micrographs, and drawing on large surfaces by tracing under low and high magnification." (2(p77)) Lantern slides continued to be widely used to photograph microscopic images through the first half of the 20th century. The slides themselves consisted of two 3.5 X 4 inch glass plates, sealed with tape to preserve the colloid on which the images were developed, and the slides were then inserted into slide holders on either side of a lantern slide projector (Figure 1, B). As such, wooden boxes used to store and transport the slides were quite heavy. To garner a few minutes of time with a prestigious lecturer, a resident or junior pathologist might offer to carry the heavy crates of glass slides to the lecture hall (S. Weiss, oral communication, November 2013).

In 1935, Kodak (Rochester, New York) produced the first 3-color, multilayer photographic film with uniquely thin emulsion layers, which made for sharp images with significantly decreased light scatter upon projection. (3) During the next several decades, universities began transitioning from using lantern slides to this color film, called Kodachrome, for photographing microscopic images for use in lectures, teaching, and patient care.

Ektachrome film, also owned by Kodak, was another commonly used film for microscopic image photography. Developed in the 1940s, Ektachrome revolutionized the photography world with its simplified processing procedure, which enabled amateurs and professionals alike to develop their own film. (4) At that time, the primary film in use, Kodachrome, required a complicated processing procedure, which was available at only a few large laboratories across America, whereas by the 1950s, many smaller laboratories were processing Ektachrome film, thus reducing turnaround times simply by reducing shipping times. Although Ektachrome film lacked the color and archival quality of Kodachrome film, it was commonly used for tumor-board image preparation from the 1950s to 1980s. (5)

Concurrent with the transition to 35-mm film slides, the method of image projection shifted from Magic Lanternstyle slide projectors, to 35-mm slide projectors. Popularized in the 1950s as a method of sharing family photo albums, these new slide projectors included novel features, such as an internal light source with requisite fan-based cooling mechanism, a condensing lens to direct light to the slide, a focusing lens for the projected image, and slide holders that could house multiple slides. (6,7) The capacity to view multiple images in sequence, along with the smaller size of the slides, further popularized microscopic image photography in academic and patient care settings.

Overhead projectors, popularized in business and educational settings during the 1950s to 1960s, worked on the same principle as 35-mm slide projectors. However, unlike slide projectors, overhead projectors projected images from transparent sheets of cellulose acetate. These images were larger than 35-mm slides, typically the size of a sheet of paper, and were placed faceup on the horizontal, lighted surface of the overhead projector and were projected onto a vertical surface. (8) Although overhead projectors had a major role in the education setting, their role in microscopic image projection was limited.

In the 1970s, Kodachrome film and processing was optimized in the form of the Kodachrome 25, a 35-mm, daylight film that could be processed overnight. (9) Before that, histology slides were not widely used in tumor board discussions, but the overnight processing enabled pathologists to bring histology images to regular multidisciplinary meetings.

In the American southeast, one of the large Kodak processing laboratories was located in the heart of Atlanta, Georgia, where optimization of color processing and reagent controls resulted in routine Kodachromes produced from the mid-1970s through the 1990s. Kodachromes from this period are still considered by some pathologists to be the highest-quality microscopic-image photographs ever produced (C. Whitaker Sewell, oral communication, December 2013) (Figure 2, A and B). Many pathologists with access to Kodachrome processing laboratories continued to use the Kodachrome film for microscopic image capturing. Conversely, the E4 film continued to be favored in areas without Kodachrome processing facilities throughout the 1970s and into the 1980s.

Many of the 35-mm films, especially Kodachrome 25, required the full light spectrum for optimal color and image quality, requiring microscope bulbs to be at maximum (or near maximum) intensity. As such, microscopes were designed with "photograph" settings that optimized image capture by 35-mm film cameras. Concurrently, mounting devices that could adapt a camera to the microscope were developed. These camera adaptors were often brand specific and were produced by the microscope manufacturers as an accessory part, available at additional cost.

Digital cameras began to gain popularity through the late 1990s and early 2000s, and microscopic image photography followed the shift from 35-mm film to digital images. Although early digital cameras lacked the resolution of Kodachromes, their increased popularity in nonmedical fields eroded the market for Kodachromes, eventually pushing Kodak to cease offering film processing in 2009. An independent film-processing facility located in Kansas continued to process Kodachrome film through 2010. (10) Similar to 35-mm cameras, digital cameras were mounted onto the top of the microscope with a scope-specific adaptor accessory.

The ease of microscopic image photography brought about by digital cameras, combined with advances in presentation technology, resulted in the transition away from slide projectors, which used 35-mm positive-image slides projected onto a screen. There were significant time-consuming and technical barriers that hindered putting words and images onto 35-mm slides for projection, and slides were rarely edited once created. The early stages of the shift to digital images coincided with the development of liquid crystal display (LCD) overhead projection devices of the 1980s. The LCD devices received video input from a computer and were able to display images from the computer monitor. When the transparent LCD device was placed on an overhead projector, an image from the computer screen could be projected vertically. (11,12) Again, this technology was not widely used for projection of microscopic image photography.

Throughout the 1980s, the use of overhead projectors became more popular in both business and academic settings, and the use of computers to create presentations increased. Although there were several precursor software programs for computerized slide composition, the Microsoft (Redmond, Washington) software program PowerPoint emerged as the forerunner in slide-editing software, ushering in new presentation styles in which slides contained many words and/or pictures that could be continually edited, updated, or reorganized. As PowerPoint gradually became the standard worldwide platform for presentations, there was increasing demand for projection devices that could easily connect to a computer and project the presentations onto a screen.

One of the first companies to address that need was Epson (Suwa, Nagano, Japan), which released its first digital projector in 1989 and a second one in 1994. These revolutionizing projectors incorporated both overhead projection capabilities and digital-panel projection into a single, relatively compact device, which helped to establish Epson as one of the leaders in digital projection technology. (13) The following decades brought development of presentation software and projection technologies, which further improved image quality and decreased both the size and the cost of digital projection devices. (12) As we stand today, PowerPoint images projected with digital projectors are the standard method for presenting material, including microscopic images.

Digital images are easy to collect and organize into large, searchable, visual libraries of pathology microscopic images, and online digital image storage and sharing sites like Flickr (; Yahoo! Sunnyvale, California) make it easy to share these images with colleagues around the world. Many microscopic images have made their way to Wikipedia (Wikipedia Foundation, San Francisco, California; and the associated Wikipedia image storage site, Wikimedia Commons (Wikipedia Foundation; where they are viewed by thousands of visitors including patients, researchers, and nonpathologist physicians. Social media sites, like Twitter (Twitter, Inc, San Francisco, California; and Facebook (Menlo Park, California;, by the nature of their infrastructure support, encourage sharing digital images. Pathology focused discussion groups, which enable members to share and discuss digital pathology images for educational purposes, have recently appeared on Facebook and are quickly growing in popularity. (14)

Another application for digital microscopic images is the Digital Pathology Diagnosis (DPDx), offered through the US Centers for Disease Control and Prevention (CDC; Atlanta, Georgia) at (accessed December 15, 2013), where pathologists, laboratory personnel, and health professionals can upload images from microscopic slides or microbiology identification tests for review by a CDC pathologist from the appropriate infectious diseases branch, and a diagnosis can be rendered. Through this service, the CDC can request material for additional or confirmatory testing of uncommon entities.

Although digital image capture helped to increase the number of applications for microscopic images, capturing the images was not necessarily any easier because digital cameras still required adaptors mounted to microscopes in addition to specialized software and hardware (eg, a computer) to capture, save, and store microscopic images.

In the late 1990s, new technology emerged for use primarily in research settings, which enabled automated scanning of whole glass slides coupled with software that could stitch together multiple images taken at various magnifications to create a composite digital image. (15) Whole slide imaging (WSI), or virtual microscopy, has advanced during the past 15 years to be a potential alternative to light microscopy in specific settings, particularly educational applications. Digital slide libraries have been used in place of light microscopes to teach basic histology in undergraduate, graduate, and medical school courses, and the technology can be used for remote consultation on unusual or rare cases. Some experts have scanned selections from their personal slide libraries and have made them available for free online. (16) Digital slides for educational purposes can be stored and shared for free through Web sites, such as PathXChange (Ventana Medical Systems, Tucson, Arizona;, Pathobin (Melbourne Accelerator Program, University of Melbourne, Parkville, Victoria, Australia;, and Whole Slide Imaging Repository (Digital Pathology Association, Indianapolis, Indiana; whole-slide-imaging-repository). Several other online resources promote sharing of whole digital slides with additional features. The Leica Biosystems (Buffalo Grove, Illinois) Aperio ePath Access provides access to a network of consultants willing to review whole slide images for an additional fee. (17) The Johns Hopkins (Baltimore, Maryland) Oncology Tissue Services will scan slides for a nominal fee and provide free storage of the digital files on their Web servers for a variable length of time, while the Aperio Technologies (Vista, California) Second Slide offers free digital storage for WSI files. (18,19)

Static Joint Photographic Experts Group (JPEG; .jpg, International Standardization Organization, Geneva, Switzerland) or Tagged Image File Format (TIFF; .tif, Aldus/ Adobe Systems, San Jose, California) digital images can be obtained from WSI by viewing a whole digital slide with one of a variety of viewing software programs. Using image-capture tools built into some viewing software (such as the Aperio ImageScope [Aperio]) or a screen-capture program (such as the Microsoft Windows Snipping Tool), a portion of the scanned slide can be saved as a static image. One benefit of obtaining static images from WSI is the ability to capture ultralow-power images of the entire piece of tissue. Large pieces of tissue are difficult to photograph in their entirety as a single image with a traditional digital camera mounted on a microscope. Incorporation of WSI into daily clinical practice continues to present several major obstacles, including up-front cost of slide scanners, immense data storage requirements, workflow modification and personnel training, integration with current laboratory information systems, necessity of a reliable network connection with high-bandwidth capabilities, and uncertainty of regulations. (20) The up-front costs alone are prohibitive to the use of WSI in smaller practices and in global health applications.

During the past decade, phones have been sold that include small cameras. Apple Inc (Cupertino, California) released the iPhone (now referred to as iPhone 2G) in 2007, which included a rear-facing, 2-megapixel (MP) camera. (21) Before the iPhone, the image capture from cell-phone cameras was limited in quality and size, without an easy way to move pictures from the phone to a computer. At that time, point-and-shoot digital cameras, which were comparable in size to the iPhone, had 2 to 3 times the pixels available to capture better microscopic images. Although the iPhone revolutionized image capture and sharing capabilities of phones, it wasn't until the iPhone 4 was released in 2010, with a 5-MP, rear-facing camera with 720p video capabilities, that the application of a smart phone for microscopic image capturing could be fully realized. (22) To our knowledge, the first report of a mobile phone camera being used to capture microscopic images through the microscope eyepiece is from Bellina and Missoni in 200936; however these authors did not include a detailed description of the technique(s) used to capture these images.

In early 2012, 2 startup companies successfully raised enough backing through the crowd-sourcing fundraiser Kickstarter (Brooklyn, New York) to begin manufacturing accessories that adapt smart phones to a microscope ocular, thus facilitating microscopic image capture via the smart phone. The SkyLight (SkyLight, Oakland, California) accessory can be used with nearly any smart phone but cannot be left on the phone as a case; it must be detached from the phone after use. (23) Comparatively, the Magnifi (Arcturus Labs, Palo Alto, California) is designed to fit iPhone models 4 or 5 as a phone case, while the ocular adaptor piece is removable. (24,25) A third company (HI Resolution Enterprises, Honolulu, Hawaii) similarly earned funds through Kickstarter in 2013 to manufacture the Snapzoom, a device designed to adapt smart phones to fit onto the ocular piece of binoculars, spotting scopes, telescopes, and microscopes. (26-28) Similar to the SkyLight, the Snapzoom is designed for use only during image acquisition and can fit a variety of smart phones with or without cases. Roy et al (29) published a report in 2014 that compared the ease of use of SkyLight, Magnifi, and Snapzoom adaptors for microscopic image capture in a clinical setting. Their experience found that Snapzoom and Magnifi provided the most streamlined use for image capture using personal smart phones in resource-limited settings or for rapid consultations. (29)

A method that uses a smart phone adaptor in conjunction with the panorama function of a smart phone camera allows for capturing "whole slide" images using a phone and microscope ocular adaptor. (30) Figure 3 demonstrates a raw image captured by the phone (Figure 3, A) and the same image after cropping the vignetting with the basic editing functions on the smart phone (Figure 3, B). Alternatively, multiple digital images can be stitched together with editing software to create a whole slide composite image. (31)

In 2013, Morrison and Gardner (32) published a detailed description of the Morrison Technique, a free-hand technique for using smart phones to capture microscopic images without the aid of additional adaptor accessories. This relatively straightforward technique is available to anyone with a smart phone and a microscope, although it requires practice, patience, and a steady hand. (33) Elston and colleagues validated the hands-free Morrison technique in 2015, comparing 2 iPhone models and a Samsung Galaxy and various microscopes (multiple Olympus models, a Leica microscope, multiple heads of a multiheaded scope, with use of polarization filters, with fluorescence microscopy, and even a stereoscopic microscope), thus demonstrating the ease of use for this technique across multiple smartphone/microscope combinations. (37) For those who lack a traditional mounted microscope camera and/or a smart phone adapter, this method may be a useful solution.

In 2013, the University of Pittsburgh Medical Center (UPMC; pittsburgh, Pennsylvania) opened an online consult service similar to the DPDx at the CDC. (34) Their service uses microscopic images taken with a smart phone using an ocular adaptor accessory. The images are uploaded directly from the smart phone to their server via an application, and a second-opinion consultation is given within 24 hours. If additional testing or material is warranted, the consult is transferred to the appropriate department for a more-traditional consultation through UPMC pathology subspecialists as appropriate. (34) By focusing the consult service on smart phone-captured images uploaded directly to their Web site, the UPMC is the first academic center, to our knowledge, to use smart phone cameras in microscopic image capture as a method for expert consultation.

A huge jump in smart phone camera image-capture capabilities was released in July 2013 by Nokia (Espoo, Uusimaa, Finland) in the form of the Lumina 1020, which spotlighted a 41-MP camera. (35) This resolution was vastly superior, compared with the 6 to 10 MP available from other smart phones at that time. Aimed at photographers, the Lumia 1020 set a new standard in the smart phone digital-photography world. Application to microscopic image capture was quickly adopted. An early 2014 publication from the Department of Pathology at the University of California Irvine detailed the superior resolution of images captured with the Lumina 1020 using the Morrison Technique, along with a unique preservation of resolution with the use of digital zoom of up to X5 original magnification (so called "lossless zoom"), a feature unique to the Lumina 1020 smartphone camera. (38) We anticipate that similar technical advances in smart phone camera technology will continue to occur in the future, potentially resulting in the eventual partial or near-total replacement of traditional digital microscope cameras with smart phone cameras for digital photomicrograph acquisition.

To compare and contrast the quality of the various digital techniques we have described, we acquired raw (not Photoshop-enhanced) photomicrographs of the same field on a hematoxylin-eosin (H&E) section of a melanoma at X200 magnification. To represent whole slide digital images, the H&E section was scanned at X20 magnification using an Aperio ScanScope (Leica, Solms, Germany) scanner in .tif file format. The field of interest was then selected using Spectrum whole slide viewer software and an image was captured via ImageScope software (Figure 4, A). To represent traditional mounted digital camera images, the H&E slide was visualized using a Nikon E400 microscope (Chiyoda, Tokyo, Japan) with a mounted Spot Flex camera (SPOT Imaging Solution, Sterling Heights, Michigan). The field of interest was captured using Spot Advanced software in .jpg file format (Figure 4, B). To represent a smart phone with adaptor microscopic photographic images, the H&E image was visualized through the ocular opening of an Olympus BX41 microscope (Shinjuku, Tokyo, Japan), and a Magnifi iPhone adapter was used with an iPhone 4S to capture the field of interest in .jpg file format (Figure 4, C). To represent free-handed (no adaptor used) smart phone microscopic photographic images, the H&E section was visualized through the ocular of an Olympus BX41 microscope, and the field of interest was captured with the camera of an iPhone 4S in .jpg file format using the Morrison Technique (18) (Figure 4, D). We invite readers to compare and contrast for themselves the quality and resolution of images obtained via these digital image-capture techniques.


In roughly 150 years, photographic technology has progressed from primitive and bulky glass-lantern projector slides to static and/or whole slide digital image formats that can now be transferred around the world in a matter of moments via the Internet. Film-based cameras, particularly those using Kodachrome film, dominated a relatively large portion of this period. The death of film and the rise of digital cameras had far-reaching effects, beyond the confines of our specialty, and this paradigm shift is perhaps one of the very best examples of how technologic innovation can completely overturn the previous status quo. Digital cameras attached to microscopes to capture .jpg and .tif images are now normal methods used in daily practice by many pathologists in the developed world. Digital microscopic images are the new standard format and have essentially replaced Kodachromes and helped to relegate film slides and slide projectors to dusty shelves in forgotten storage rooms of pathology departments everywhere. It is likely that many current residents have never seen a Kodachrome or a slide projector. Given the relative low cost, widespread availability, and ease of use, we suspect that smart phone cameras are poised to potentially turn the pathology image-acquisition status quo on its head yet again for the second time in 2 decades; however, only time will tell.

Please Note: Illustration(s) are not available due to copyright restrictions.


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Annie O. Morrison, MD; Jerad M. Gardner, MD

Accepted for publication March 17, 2015.

Published as an Early Online Release May 19, 2015.

From the Department of Pathology & Laboratory Medicine, Emory University School of Medicine, Atlanta, Georgia (Dr Morrison); and the Department of Pathology, University of Arkansas for Medical Sciences, Little Rock (Dr Gardner). Dr Morrison is now located at Cockerell Dermatopathology, University of Texas Southwestern, Dallas.

The authors have no relevant financial interest in the products or companies described in this article.

Reprints: Annie O. Morrison, MD, Cockerell Dermatopathology, University of Texas Southwestern, 2110 Research Row, Suite 100, Dallas, TX 75235 (e-mail:

Caption: Figure 1. Historic microscopic image-capturing devices. A, Leitz Edinger Apparatus, circa 1910, used to align a Lantern Slide camera to a microscope, B, Bausch and Lomb Carbon Arc Balopticon C Lantern Slide Projector, circa 1908, used to project images from Lantern Slides. (Photographs courtesy of Bruce Allen).

Caption: Figure 2. Examples of Kodachrome (Kodak, Rochester, New York). A, Slide. B, Image (scanned from slide), circa 1978. (Slide and slide image courtesy of C. Whitaker Sewell, MD).

Caption: Figure 3. Examples of "whole slide scanning" using the smart phone panoramic function A, Raw image captured. B. Image cropped using basic smart phone editing functions.

Caption: Figure 4. Melanoma captured with whole slide digital scanning (A), camera mounted on microscope (B), smart phone camera using eyepiece adaptor (C), and smart phone camera using free-hand technique (D) (hematoxylin-eosin, original magnification X200 [A through D]).
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Author:Morrison, Annie O.; Gardner, Jerad M.
Publication:Archives of Pathology & Laboratory Medicine
Date:Dec 1, 2015
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