Carl Zeiss MicroImaging AIS. Digital Pathology The Future. Andrew Lesniak Director, Product Management
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1 Carl Zeiss MicroImaging AIS Digital Pathology The Future Andrew Lesniak Director, Product Management
2 Defined Industry Needs Computer Aided Imaging and Analysis will play a key role in the increase of Diagnostics by 2010 A 60% growth factor Leverage Core Zeiss Strengths of Innovation, Advanced Imaging and Quality
3 Product Constellation Work Load Management Assigned cases Signed out cases Shared cases Slide Viewing & Management All slides associated with case Report Generation & Clinical LIS Integration Referencing Collaboration & Presentation Quantitative Histopathology Scoring & Analysis Infrastructure Live Slide Imaging Fluorescence Spectral Imaging Optical Volume Scanning Gross Samples Imaging High Speed Image Acquisition
4 Making it Possible: Multi-channel Fluorescence TexasRed Multi channel Fluorescence Image acquisition in several individual fluorescence channels FITC Overlay DAPI
5 Scanning Advancements: Multiplexing Fluorescence Acquisition
6 Digital Pathology Workflow Enabling Scale Faster to change slides digitally than physically Enable organization of imagery by animal treatment group Side by Side Comparison Determination of No Effect versus Minimal Effect Rapid access to alternate staining modalities / Serial Sections Can be linked to internal LIMS / Histology systems Successful integration with multiple vendor imaging solutions SSL Secure Communications, Auditing and User Access Control Lists
7 Digital Pathology Workflow Enabling Scale Minimum Requirements (Web Server): - Dual Xeon 3.0GHz+ (or equivalent) processor, 2GB - 10k (or greater) RPM Disk Storage - Windows Server 2003 Standard Edition w/iis Installed - SQL Server 2000 Enterprise Edition PDIM Distributed System (or MSDE) - Apache Tomcat v5.5 Image Transfer LAN / WAN Backbone Web-based Image Viewing (Client)
8 Digital Pathology Workflow Scalable Platform Hierarchical Folder Management Automated grouping of Slides by Attributes, such as Study ID As Images are created, Easy to manage and deploy Security of Images by User individual user logins can be assigned to a given folder for multi-client use. Replication Utilized by Pharma / Biotech to engage Collaborators Synchronization of internal and external systems via copy / publish methods.
9 Digital Pathology Workflow Enabling Scale
10 Leveraging a Leading Product Portfolio for a Complete Pathology Value Add Solution Image Repository Framework Toxicology Desktop Digital Pathology Classroom Histotech Workbench Cytotech Workbench Anatomic Pathology Desktop Cytopathology Desktop
11 Digital Pathology Workflow: Enabling Scale Workflow, Educational and Reference Materials Virtual Lab creation- organization of Digital Images by context Linking a slide to relevant accession, histological and pathological context information Linking areas of interest on a slide to relevant teaching or lesion specific information Preservation of compound related effects for future reference
12 Digital Pathology Workflow Enabling Scale Study organization by Tissue Type, Animal or Dosage Group. Slide review progression by sequential animal, organ or dosage for rapid and structured review of tissues. Per tissue findings recorded in database. Color-coding for heads-up slide status. Grouping and rapid access to slides for comparison of tissues marked as Review or Abnormal for rapid revisit of lesions and/or peer review / PWG discussion.
13 Digital Pathology Workflow Enabling Scale Side by side tissue comparison for accurate assessment of lesions. List based access to tissues grouped for Review and/or Abnormal enables the ability to sort tissues by pathology grade and then rapidly compare with control to determine severity of lesion. Next / Previous slide access navigates each slide column columns can represent the series for an animal, tissue type or tissues marked to be reviewed.
14 Digital Pathology Analysis: Innovations with Imaging Tissue Scoring and Quantitative Analysis Membrane Stains Nuclear stains Integrated Optical Density Model Based Analysis Liver, Kidney, Muscle Rare Event Detection Sentinel Lymph nodes Bone marrow Object Detection and Counting Nuclear Cell Counting, Micro-Vessel Density, Ploidy Tissue Micro Array Scoring
15 Quantitative Analysis Examples Liver Steatosis Method Key observation = the accumulated fat that characterizes steatosis is seen as empty space in the processed tissue section. 1. Color modeling allows for Hematoxylin, Eosin and the lack thereof to be robustly calculated. White Compartment (no overlay color) Nuclear Compartment (blue overlay) Eosinophillic Compartment (red overlay) 2. Once these compartments have been identified and their areas measured, the percent of white space is used as an approximation of % steatosis: % steatosis % white space = ~ 100 x (white area) (white area) + (blue area) + (pink area)
16 Basic Object Differentiation Comparison with Standard Control Sample KS p-value Control/Normal Treated
17 Advanced Object Modeling Comparison with Standard Control Control Treated Histograms of white space eccentricity (roundness).
18 Advanced Object Modeling Comparison with Standard Control Sample Control Mean +/- S.D. Treated Mean +/- S.D Circular White Objects / /- 48 Total White Objects / /- 350
19 Quantitative Analysis Examples Kidney Nephropathy
20 Quantitative Analysis Examples Kidney Nephropathy Specimen Kidney Analysis Research goal Identification of Glomular Tuft Area, Mesangium Area and Nucleation 1. Scanned images are viewed in their entirety. 2. Areas for analysis are screened based on properties. Method Automated Assessment of Glomeruli structures further refinement of cellular staining in an automated workflow protocol. 3. Review for quality, and exclude as needed 4. Numerical results are provided.
21 Quantitative Analysis Examples Cell Classification in Cytospin Specimen Immunocytochemically stained leukocytes in blood or bone marrow cytospin. Research goal Discriminate large vs small cells and immunocytochemically positive vs negative cells. Challenge: Must make several discriminations among the included populations. Exclude erythrocytes Separate small (lymphoid) from large (myeloid) cells Distinguish immunocytochemically positive and negative cells
22 Quantitative Analysis Examples Cell Classification in Cytospin Discriminations such as these are readily performed through flow cytometry: Monocyte Neutrophil Eosinophil Lymphocyte Basophil Erythrocyte RBC
23 Quantitative Analysis Examples Cell Classification in Cytospin To successfully classify the individual identified objects, we must make use of their extracted features: Size Hematoxylin Chromagenic Red blood cells small none/low medium/high ICC-negative lymphoid cells small high none/low ICC-positive lymphoid cells small high medium/high ICC-negative myeloid cells large high none/low ICC-positive myeloid cells large high medium/high
24 Molecular and Digital Pathology A strong couple Digital Pathology from Carl Zeiss Molecular Pathology from Carl Zeiss MiraxScan & MiraxDesk & Mirax Micro Automated digital pathology Up to 300 slides in one run (MiraxScan) Virtual slides are available world wide at any time PALM MicroBeam Contact-free and against gravity = contamination free Molecular downstream analysis (DNA, RNA, Proteins) Automation can be included (automated object recognition and collection)
25 PALM MicroBeam Capture Microdissection at your finger tips: New user interface Simple Convenient Well structured Laser management on the left Microscope management on the right Drawing tools at the bottom Navigator, Information Center and Element List included Optional Modules: Extended Focus (EF) Multi channel fluorescence Time lapse Database AxioVision Commander (Scripting)
26 PALM MicroBeam Capture Laser Microdissection from membrane mounted tissue Membrane Mounted Tissue Laser Microdissection from glass mounted tissue Mounted Tissue Object Slide PALM MembraneSlide Preserves tissue morphology Enables dissection of any shape & size Facilitates ablation & tissue separation Allows fixation & staining Object Slide Glas Mounted Tissue Can be used on archival material Homogenizes the tissue Unique PALM feature
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