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