Carl Zeiss

MicroImaging AIS

Digital Pathology

The Future

Andrew Lesniak

Director, Product Management

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

Defined Industry Needs

Work Load

Management

•Assigned cases

•Signed out cases

•Shared cases

Slide Viewing & Management

•All slides associated with case

Live

Slide Imaging

Optical

Volume

Scanning Gross

Samples

Imaging

High Speed

Image

Acquisition

Fluorescence

Spectral

Imaging

Quantitative

Histopathology

Scoring &

Analysis

Referencing

Collaboration &

Presentation

Report Generation

& Clinical LIS

Integration

Infrastructure

Product Constellation

� Multi channel Fluorescence

Image acquisition in several

individual fluorescence channels

DAPI

Overlay

TexasRed

FITC

Making it Possible: Multi-channel Fluorescence

Scanning Advancements: Multiplexing

Fluorescence Acquisition

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

PDIM Distributed System

LAN / WAN Backbone

Web-based Image Viewing (Client)

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

(or MSDE)

- Apache Tomcat v5.5

Image Transfer

Digital Pathology Workflow

Enabling Scale

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.

Digital Pathology Workflow

Digital Pathology Workflow

Enabling Scale

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™

Workflow, Educational and Reference Materials

�Virtual Lab creation- organization ofDigital Images by context

�Linking a slide to relevant accession,histological and pathological contextinformation

�Linking areas of interest on a slideto relevant teaching or lesion specificinformation

�Preservation of compound relatedeffects for future reference

Digital Pathology Workflow:

Enabling Scale

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.

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.

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

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.

2. Once these compartments have been identified and their areas measured, the percent of

white space is used as an approximation of % steatosis:

White Compartment

(no overlay color)

Eosinophillic

Compartment

(red overlay)

Nuclear Compartment

(blue overlay)

100 x (white area)

(white area) + (blue area) + (pink area)% steatosis % white space = ~~

Quantitative Analysis Examples

Liver Steatosis

0.0008“

0.0812“

0.0110“

0.0423“

0.1073Treated

0.0147“

0.0685“

0.0017“

0.1628“

0.1073Control/Normal

KS p-valueSample

Basic Object Differentiation

Comparison with Standard Control

Control Treated

Histograms of white space eccentricity (roundness).

Advanced Object Modeling

Comparison with Standard Control

1037 +/- 350197 +/- 48Mean +/- S.D

855179“

1239257“

966175“

610139“

1515235Treated

168 +/- 9235 +/- 19Mean +/- S.D.

9820“

18231“

17034“

31268“

7923Control

Total White ObjectsCircular White ObjectsSample

Advanced Object Modeling

Comparison with Standard Control

Quantitative Analysis Examples

Kidney Nephropathy

Specimen

Kidney Analysis

Research goal

Identification of Glomular Tuft Area,

Mesangium Area and Nucleation1. Scanned images are

viewed in their entirety.

2. Areas for analysis

are screened based

on properties.

3. Review for quality, and

exclude as needed

4. Numerical results

are provided.

Method

Automated Assessment of

Glomeruli structures – further

refinement of cellular staining in

an automated workflow protocol.

Quantitative Analysis Examples

Kidney Nephropathy

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

Quantitative Analysis Examples

Cell Classification in Cytospin®

Neutrophil Eosinophil

Basophil

Erythrocyte

Monocyte

Lymphocyte

RBCRBCRBCRBC

Discriminations such as these are readily performed through flow cytometry:

Quantitative Analysis Examples

Cell Classification in Cytospin®

Red blood cells

ICC-negative lymphoid cells

ICC-positive lymphoid cells

ICC-negative myeloid cells

ICC-positive myeloid cells

Size Hematoxylin Chromagenic

small none/low medium/high

small high none/low

small high medium/high

large high none/low

large high medium/high

To successfully classify the individual identified objects, we must make use of

their extracted features:

Quantitative Analysis Examples

Cell Classification in Cytospin®

Molecular and Digital Pathology

A strong couple

Digital Pathology from Carl Zeiss

MiraxScan & MiraxDesk & Mirax Micro

Molecular Pathology from Carl Zeiss

PALM MicroBeam

• Automated digital pathology

• Up to 300 slides in one run (MiraxScan)

• Virtual slides are available world wide at

any time

• Contact-free and against gravity =

contamination free

• Molecular downstream analysis (DNA,

RNA, Proteins)

• Automation can be included (automated

object recognition and collection)

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)

PALM MicroBeam Capture

Laser Microdissection from

glass mounted tissue

Object Slide

Mounted Tissue

Glas Mounted Tissue

• Can be used on archival material

• Homogenizes the tissue

• Unique PALM feature

Mounted Tissue

PALM MembraneSlide

Laser Microdissection from

membrane mounted tissue

Object Slide

Membrane

• Preserves tissue morphology

• Enables dissection of any shape & size

• Facilitates ablation & tissue separation

• Allows fixation & staining

PALM MicroBeam Capture

Thank You!!

Questions?