Standardization in HER2 Testing –

A UK Reference Centre Approach

Michael Gandy, UCL-AD, Copenhagen 2011

Background

Background

Rockefeller Building

Complex Testing

IHC In Situ

Hybridisation

Molecular

Pathology

Digital

Pathology

IHC Assays CISH/FISH/SISH Assays Genetic AnalysisDigital Pathology & Image Analysis

Aims

•Look at the varying nature of sample

types we test for HER2.

•Look at the testing methods available

and what standardization can mean.

•Describe some of the factors which

effect the end result

•Breast Cancer

•Most Common Cancer in the UK

•2008 – 47,700 women (1 in 8)

340 men

•1.38 million worldwide

www.cancerresearchuk.org

Patient Demographics

Patient Demographics

www.cancerresearchuk.org

Patient Demographics

www.cancerresearchuk.org

Sample Types –

Breast & Gastric Cancer

Sampling – Cytology FNA

Sampling – Core Biopsy

Sampling – Lumpectomy/WLE

Sampling – Mastectomy

Sampling – Sentinel Lymph Node

Gastric Cancer

Standardization

Standardization

Quality

Consistency

Factors Affecting Standardization

•Wolff et al 2007 ASCO/CAP HER2 Testing Guidelines

Good, Bad – Why?

Pre-Analytical Factors – Combination

Factors Affecting Standardization

Appropriate Controls

Appropriate Controls

Factors Affecting Standardization

Technical Considerations

Inking Margins

Technical Considerations

Z-axis

Depth of Field

Technical Considerations

Overlapping nuclei

Over Digestion

Technical ConsiderationsDAPI Counterstain

Technical Considerations

Inadequate Stringency Wash – Fine Deposit

Technical Considerations

Inadequate Stringency Wash – Fine Deposit

Technical Considerations

Inadequate Stringency Wash - Trapping

Technical Considerations

Inadequate Stringency Wash - Trapping

Technical Considerations

Inadequate Stringency Wash - Glass

Technical ConsiderationsAutofluorescence

Technical ConsiderationsFilter Sets - Excitation & Emission Wavelengths

HER2 ISH Testing – Breast &

Gastric Cancer

Lymphocytes

Stromal cells

Epithelium

Interesting Cases

Case 1 – H&E

Case 1 – IHC

Case 1 – ISH

Case 2 – H&E

Case 2 – IHC

Case 2 – IHC

Digital Imaging

Digital Imaging

Digital Imaging

Take Home Message

Standardization aim to consider

the whole process. Combination of

small factors can have a similar

effect to one large factor.

Take Home Message

Many Thanks...

...Any Questions