Poster Session

s of the 1st Congress of the International Academy of Digital Pathology: Poster Session 199 Medical School Pathology education supplemented with web-based virtual microscopy Rajyasree Emmadi, Amy Y. Lin, and Andy V. Pham Department of Pathology, University of Illinois at Chicago (UIC), IL, USA Background: The year 2 Medical student Pathology practicum at UIC, designated Small Group Discussion (SGD), comprises 184 students in groups of 14–16 each. Traditionally the practicum has involved students examining and learning morphologic pathology by viewing glass slides through the microscope, with accompanying instruction, in 2-hour sessions. We decided to introduce web-based virtual microscopy to the SGDs to augment teaching and facilitate self-paced student learning. Methods: Selected glass slides were scanned using the Aperio ScanScope (Aperio, Vista, CA), to obtain the virtual slide images and interactive cases were created using Digital Slide Box (DSB) software (SlidePath, Dublin, Ireland). Each virtual slide was annotated with key microscopic features of the case. Each case, in turn, was accompanied by a narrative, which included a clinical history, physical examination fi ndings, and gross and microscopic descriptions. A total of 39 virtual slide sets were utilized for the current academic year. Hyperlinks within the narrative are available to integrate clinical photographs, gross photographs, imaging studies, multimedia, additional virtual slides or slide annotations to the case. At the end of each module, self-assessment quizzes helped students test their understanding and identify individual weaknesses. Results: These interactive, web-based virtual microscopy cases were provided to the M2 students the week prior to the actual SGD. This allowed the students to study the material at leisure and without a time constraint. At the actual SDG, the students were able to concurrently view different glass slides of the specifi c disease process as well as re-review the online material. While initially the in-class bandwidth slowed the online review noticeably, this was resolved by effecting a change in the DSB server confi guration. An informal feedback showed that the students were very receptive to this new technology and found these selfstudy cases useful in increasing their recognition and understanding of pathologic processes in diseases. They particularly liked the ‘anytime, anywhere’ access and the ability to dispense with the microscope. Conclusions: An interactive, web-based virtual microscopy case study set makes Pathology more accessible and inviting to Medical students. In the next academic year we plan to expand the virtual microscopy content. Toward an annotated digital Multiphoton Microscopy (MPM) histology atlas of fresh human bladder biopsies for intra-cystoscopy guidance in bladder cancer diagnosis Sushmita Mukherjee, Manu Jain, Brian D. Robinson, Joshua Sterling, Douglas S. Scherr, Bekheit Salmoon, Frederick R. Maxfi eld, Warren R. Zipfel and Watt W. Webb Department of Biochemistry, Weill Cornell Medical College, New York, NY, USA 2 Department of Urology, Weill Cornell Medical College, New York, NY, USA 3 Department of Pathology and Laboratory Medicine, Weill Cornell Medical College, New York, NY, USA 4 Department of Biomedical Engineering, Cornell University, Ithaca, NY, USA 5 School of Applied and Engineering Physics, Cornell University, Ithaca, NY, USA Background: Hematoxylin and eosin (H&E)-stained sections obtained from formalin-fi xed specimens is the current gold standard for histopathological diagnoses. While these methods are highly reliable, they have lengthy time requirements (processing, sectioning, staining, and reading by pathologists). Although efforts at digitization of glass slides are in progress, most pathologists still read analog slides; therefore, automated morphometry and real time online consultations are rare. Furthermore, the 2-dimensional nature of histology slides precludes assessment of 3-dimensional tissue architecture without time consuming serial sectioning. Methods: Multiphoton microscopy (MPM), a nonlinear imaging technology, generates 3-dimensional histology of the tissue at sub-cellular resolution and at depths up to 0.5 mm below the tissue surface. This allows nearly instant imaging of fresh (unfi xed, unsectioned, and unstained) tissue based on spectrally resolved intrinsic tissue emission (ITE) signals: (1) autofl uorescence from cell cytoplasm components and 200 Abstracts of the 1st Congress of the International Academy of Digital Pathology: Poster Session elastin fi bers; and (2) Second Harmonic Generation (SHG), a nonlinear scattering signal from collagen bundles and oriented microtubules. Using a single excitation wavelength and collecting emission signals using wavelength bandpass fi lters, SHG and various autofl uorescence components can be separately acquired, analyzed and color-coded for easy visualization. Results: We analyzed ex vivo tissues from human bladder biopsies by MPM and compared our diagnostic impressions with gold standard hematoxylin and eosin stained sections of the same specimens. MPM images alone provided suffi cient details to classify most lesions as either benign or neoplastic, using the same basic diagnostic criteria as histopathology, namely architecture (fl at or papillary) and cytologic grade (benign/low grade or high grade). We have begun the generation of a pathologist-validated and annotated digital atlas containing both the MPM image sets and the corresponding H&E histopathology. Conclusion: A validated digital MPM atlas may provide intra-cystoscopic guidance to urologists in situ. Spectral sensing method for practical use Yasuhiro Fukunaga, Saori Shimizu, Kensuke Ishii, and Kosei Tamiya Olympus, Inc., Japan A pathology imagery interpretability rating scale for virtual microscopy Peter Kragel and Bill Oliver Department of Pathology and Laboratory Medicine, East Carolina University, Greenville, NC, USA Automated 3D-reconstruction of histological sections Maristela L. Onozato, Mark Merren, and Yukako Yagi Department of Pathology, Massachusetts General Hospital, Boston, MA, USA Introduction: Three-dimensional (3D)-reconstruction from serial sections is a valuable tool to provide structural and morphometric data of complex structures however, unlike virtual sections such as from radiology scanned images, 3D-reconstruction from physical histological sections are more challenging due to the distortion that occurs from processing and positioning each section on each individual slide. Objective: In this work we tested whether a completely automated tissue processing system with automated sectioning machine and slide scanning system could generate precise 3D-reconstructions of tissues that could match the images obtained with an optical tomographic device. Methods and results: Human lung and heart and rat kidneys were scanned with large-fi eld-optical coherence tomography (LF-OCT, LLtech Inc., Princeton NJ) in order to obtain micrometer resolution images of unprocessed biopsy tissues. The tissues were then embedded in paraffi n and sectioned with Kurabo-Automated tissue sectioning machine. Serial sections were then automatically stained and scanned with a Whole Slide Imaging device. Images were 3D-reconstructed with inbuilt software and compared with the unprocessed images obtained with LF-OCT. 3D-reconstructed images of human lung and heart showed a very close structural details to the images obtained with LF-OCT. 3D-reconstructed rat kidney revealed details of spatial distribution and structural interaction of the nephron different segments including the vascular pole of the glomerulus however some minimal distortion could not be prevented by this all-automated system. Conclusion: Technology advances are allowing simpler ways of obtaining 3D images from 2D slides therefore a better interpretation and analysis of complex structures such as the nephron with details that can not be provided by light optical imaging systems however still some congruency details and imaging corrections have to be implemented. Automated sectioning machine for paraffi n blocks Maristela L. Onozato, Stephen Hammond, Mark Merren and Yukako Yagi 1 Pathology Information and Communication Technology (PICT) Laboratory, Department of Pathology, Massachusetts General Hospital, Boston, MA, USA 2 Department of Pathology, Boston University, Boston, MA, USA Abstracts of the 1st Congress of the International Academy of Digital Pathology: Poster Session 201s of the 1st Congress of the International Academy of Digital Pathology: Poster Session 201 Background and signifi cance: Microtomy has been a limiting factor for the development of a fully automated system for tissue histology. In this work we present a novel robotic paraffi n-sectioning machine and we compare the automated sectioned slides with traditional manual sections. Material and methods: A total of 46 blocks were manually or automated sectioned at 4 μm and then hematoxylin-eosin (H&E)-stained. Sections were scored by two blinded-professionals based on the presence of technical imperfections or irregularities that could interfere with pathology evaluation. The score ranged from1 to 4, with scores closer to 0 refl ecting perfect sections. Immunohistochemistry was also performed in breast tissue to confi rm that antigenicity was not affected by the procedure. Results: Automated sectioned slides showed remarkable quality compared with manual sections with fewer imperfections (automated score 0.87 ± 0.07 vs. manual 1.49 ± 0.07, p < 0.001) however manual sectioning for the 46 blocks was almost two-fold faster than the robotic system. The robotic-system showed the best performance for serial sections with slides showing stable thickness and same orientation allowing easy stacking and 3D reconstruction. Breast tissue showed preserved antigenicity demonstrated by immunostaining for human epidermal growth factor re