Session: Innovative Technologies

Abstract Title: Tissue Microarray Technology:

New opportunity for Tissue Banks to contribute

to Cancer Research and Diagnostics.

Pasquale De Blasio • Founding President of ESBB

• Member of Marble Arch Int. Biobanking Working Group

• President of ISENET-USA LLC

Combines tens to hundreds of paraffin-

embedded tissue specimens into a

single paraffin block.

Tissue Microarray Technology

Pathology archives PPFT Tissue Blocks & Slides Tissue Microarrayer

TMA Block

• It is possible to slice up to 200 consecutive

sections of 4-5μm from each TMA block,

mounted on a microscope slide

• The TMA slide can be processed like

ordinary tissue sections with a wide range

of techniques: Histo

staining, Immuno

histo

Immuno

fluorescent.

FISH

Tissue Microarray Technology

“Advantages to use

Pathology Archives”

• widest collection of clinical tissues

available with the entire clinical

heterogeneity range

• Tissue samples are related to clinical

records and have follow-up and

outcome information

• Are under the responsibility of

pathologists that have access to all the

information and are bound by

professional secrecy

• It is now possible to perform many

types of molecular analysis on this

type of tissues.

Tissue Microarrays

Validation In Real Patient Samples

In Vitro

Screening

Methods

Clinical

Trials

Epidemiology

Based Studies

Multi-tumor

Systems

Micro-Arrays Other

Methods

TMAs in Biomarker Development

TMAs advantages: • the speed of molecular analyses is increased by more than 100-fold.

• precious tissues are not destroyed

• large number of molecular targets can be analysed from consecutive

TMA sections (Cost reduction). Compliments of S. Hewitt, NCI TARP Laboratory

First scientific pubblications on TMA

From Tissue To Arrays

• Develop Hypothesis

• Identify Tissue

• Prepare Tissue

• Array Tissue

• Section Arrays

• Perform Experiment

• Analyze Data

• Publish

• Repeat

Compliments of S. Hewitt, NCI TARP Laboratory

Quality of Tissue is everything

1

2

3

4

5

6

7

Tissue Quality

Histology

Proteins

Nucleic Acids

Diagnostic Reporting

Clinical Annotation

Storage Of Biomaterial

Compliments of S. Hewitt, NCI TARP Laboratory

Challenges in the

Construction of a TMA

• Identifying Adequate/Appropriate

Material

• Negotiating MTA/IRB Approval

• Specimens Present Special

Challenges

– Biopsy & Small Specimens Are Hard

To Array

– Prior Use of Tissue for diagnostic

• Diversity of Specimen Sources

– Regional Differences In Diagnosis &

Treatment

– Differences In Specimen Handling

• Informatics

• Managing Expectations

– Pathologist Vs Scientist

TRACEABILITY

Integrated Tissue MicroArray

Pathology Platform “workflow”

Clinical Database

Quantitative Digital

Pathology

Digital Scanners

Fish Fluorescence Histo Immuno

Tissue Microarrayer

Platform

Summary

Harvest and re-suspend cells in agarose

Embed cells in paraffin donor blocks

Pick cell or tissue cores from donor blocks

FFPETs donor blocks

Spot cell/tissue cores in recipient paraffin blocks

Immunohisitochemical/ immunofluorescemnce

analysis

DNA and RNA extraction

Place cell/tissue cores in specific

vessels

• Tissue microarrays (TMAs) have become a

mainstay in preclinical and translational research,

especially for the development of biomarker assays

for characterization of disease.

• They allow for analysis of extremely small amounts

of tissue, thus preserving valuable tissue blocks.

• At the same time, they dramatically increase the

efficiency and cost–effectiveness of performing

tissue-based studies by enabling the examination of

10s to 100s of different patient samples on the same

slide.

Summary

Summary

• A block of Samples from

hundreds of Tissue Blocks

– Multiple Samples (3-4) from

Paraffin Embedded or Frozen

Tissue blocks

– Arranged in an organized way

Platform For High-Throughput Pathology

Immuno Histo Fluorescence Fish

Compliments of S. Hewitt, NCI TARP Laboratory

Summary

• A Protein Array with Retained

Histomorphology

– Cell Type Localization

– Cell Type Quantification

– Subcellular Localization

Platform For High-Throughput Proteomics

Immuno Histo Fluorescence Fish

Compliments of S. Hewitt, NCI TARP Laboratory

H&E IHC IHC (75 cases, dupl)

LABOR INDIRECT

COSTS

MATERIALS

SINGLE SLIDES TMA

€ 2

5

€ 2

5

€ 1

,88

8

€ 3

8

€ 3

8

€ 2

,81

3

€ 2

71

€ 4

€ 2

71

€ 0.00

€ 500.00

€ 1,000.00

€ 1,500.00

€ 2,000.00

€ 2,500.00

€ 3,000.00

Total cost per unit Cost per patient Cost per 75 patients

H&E

IHC

IHC on TMA

Conclusion

Compliments from G. Cattoretti, UNIMIB- Italy

Summary

Brest, PgR (DAB),

Pos Rat (N, A)

Brest, Gult1, IHC,

Membrane (A, I)

Connectivity

Melanoma

KI-67 + MART1

(DAB, AEC) Proliferation

Courtesy of: Giorgio Cattoretti

Associate Professor, Universitá degli Studi Milano-Bicocca

Director, Division of Surgical Pathology, Cytology, Medical Genetics and Nephropathology

Azienda Ospedaliera San Gerardo, Monza

This material may be copyrighted. Do not diffuse without authorization.

http://www.tma.cattoretti.it/

Conclusion

Compliments from G. Cattoretti, UNIMIB- Italy

Tuesday 21st October

• Training Course on Tissue Microarrays and

Quantitative Digital Pathology.

• Topics covered include:

– planning a biomarker research project using TMAs,

– diagnostic use of TMAs

– quantitative digital pathology analysis.

– Includes a hands-on session to make your own TMA

and analyse your slides (bring your own blocks and

slides).

Acknowledgments

Maurizio Andrea Aikaterini Tatiana Davide

Monica Simona Alberto Alessandria Valeria