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2679                          Clinical Validation of High Throughput AQUA and Multispectral Imaging for Breast Cancer     ...
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Clinical validation of high throughput AQUA and multispectral imaging for breast cancer


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This poster describes the evaluation of PerkinElmer's Vectra2 Automated Multispectral Slide Analysis System in a clinical environment with a focus on quality and pathologist involvement. (Poster presented at AACR 2013).

Cristen K. Hays (1), Kerry Fitzgerald (1), Bana Mouwakeh (1), Peter Miller (2), Clifford Hoyt (2), Jason Christiansen (1) and Danielle Murphy (1).

1 - Genoptix Medical Laboratory, Carlsbad, CA
2 - Perkin Elmer, Hopkinton, MA

For further information on PerkinElmer’s Microscopy Imaging Systems and Software, as presented in this poster, please visit

Published in: Health & Medicine
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Clinical validation of high throughput AQUA and multispectral imaging for breast cancer

  1. 1. 2679 Clinical Validation of High Throughput AQUA and Multispectral Imaging for Breast Cancer Cristen K. Hays , Kerry Fitzgerald , Bana Mouwakeh , Peter Miller , Clifford Hoyt , Jason Christiansen , Danielle Murphy 1 1 1 2 2 1 1 1 Genoptix Medical Laboratory, Carlsbad, CA; Perkin Elmer, Hopkinton, MA 2 INTRODUCTION Background of Vectra 2 and Vectra Review Stability and Reproducibility of Instrumentation Concordance of AQUA Scores Generated by New Platform The high rate of discordance currently identified in the scoring of breast cancer biomarkers ER, PR, Her2, and Figure 1. The Vectra 2 platform is a multispectral imaging A. Figure 5. A. We system for acquisition and analysis of brightfield/ ER PR used ROC analysis Ki67, which directly impact patient management, drives a fluorescent microscopy with tissue microarray (TMA) to investigate the and whole tissue sections (WTS). It relies on associated concordance of Vectra growingclinicalneedforthedevelopmentofdiagnostictests software, Inform and Nuance, to generate algorithms 2 generated AQUA which offer enhanced reproducibility while maintaining used to identify regions of interest (ROI) at 4X for further scores for ER, PR, sampling at 20X while reducing background fluorescence and Her2 based off of accuracy in a high throughput environment. We currently for pure imagery. Images are easily reviewed with Vectra validated cutpoints. For offer AQUA® technology in our clinical lab for breast Review software. Vectra Review displays a stitched image Ki67 we used linear of the entire tissue section and allows the reviewer to regression analysis cancer. A significant limitation to using this technology toggle between 4X and 20X images in order to accept or and observed a high within a diagnostic laboratory is low throughput and reject images based on quality prior to scoring. B. correlation of the Vectra platform to our manual annotation of patient invasive tumor. To address current system. this, we evaluated the Vectra2™ automated multispectral slide analysis system in a clinical environment with a focus on quality and pathologist involvement. Materials and Methods Her2 Ki67 All results were generated using de-identified clinical C. samples. The Vectra system was used to acquire Figure 3. AQUA technology combined with the Vectra platform provides a stable and reproducible system for breast biomarker expression in invasive tumor regions AQUA analysis. A. Scoring is dependent upon changes scored by AQUA technology. The multi-step process in light output, use of a FluorRef slide allows for consistency in score reporting independent of changes of image acquisition was assessed in conjunction with in light output. B. Repeated scanning does not result the reproducibility of region selection for scoring as in significant changes in AQUA scoring in (left) TMA, p=0.4972 or (right) WTS ~9%CV, n=7. C. Imaging determined by invasive cancer tissue classifier algorithms of WTS results in highly reproducible AQUA scores developed through the use of Inform and Nuance between four different Vectra 2 machines p=0.7335. applications. We also developed a review application to allow remote access to images prior to reporting of Vectra 2 Platform Improves Efficiency for Automated Quantitative Analysis Development of Robust Software Parameters patient results. Figure 2. Introduction of the Vectra A. 2 platform into the clinical lab Results A. offers a significant improvement to Conclusions workflow for performing AQUA. A. Automated QUantitative Analysis Normalization of system intensity compensation produced (AQUA) is used to quantitatively We show that the Vectra 2 platform is as accurate as our currently clinically validated platform for intensity values approaching unity. Algorithms defining measure biomarker expression AQUA scoring of ER, PR, Her2, and Ki67. The introduction of advanced tissue-finding algorithms by assessing marker presence invasive tissue score were found to be highly correlative (target) within a defined tumor concisely recognize relevant tissue regions unique to each biomarker without manual annotation (ER, PR, Her2 AUC 0.966-0.967; Ki67 R2) to the current mask. B. The current workflow and greatly improved the clinical workflow. This deployment of digitalized pathology resulted involves review of a hematoxylin clinical platform with pathologist annotation. Together, B. and eosin (H&E) section to in a reduction of time to generate scores, which marks a significant advance for our clinical lab. these features resulted in acquisition of conserved regions determine invasive tissue ROI for Overall, this clinical validation demonstrated that the AQUA-paired Vectra 2 system is a highly fluorescent immunohistochemistry of invasive tumor in a highly reproducible fashion, such (FIHC) section. A FIHC section robust, reproducible, and high throughput advance to our currently available diagnostic breast that tissue scores were the same for the same sample on is scanned and annotated before cancer platform that still allows for clinical lab and pathologist review. AQUA scoring and includes multiple days and with different instruments, p=0.7335. several manual steps. Use of • AQUA technology and the stability of the Vectra 2 platform enables high reproducibility of Scores were consistent amongst varying parameters, Vectra eliminates the need for AQUA scores between Vectra machines and during repeated scanning. H&E staining before scoring, while including repeated scanning (~9 %CV, n=13 runs) and utilizing tissue-finding algorithms differing field sampling parameters while decreasing time to identify ROI and reduce the • User-defined tissue algorithms of the Vectra 2 system enable robust HPF collection and to 15 min per slide for processing with parallel analysis. number of high powered images AQUA scoring with clinical relevance. captured. The time to acquire and B. Figure 4. A. User-defined ROI finding Our development of the Vectra Review software enables analyze a specimen is ~15 to 20 algorithms reliably identify the same regions • Vectra scores are highly correlated with those of our current clinical platform. minutes. A pathologist reviews during repeated scanning on the same and visualization of regions selected for biomarker scoring different machines. B. Score generation with that can be reviewed by clinical staff remotely, while not images with the associated H&E varying numbers of high powered field images • Vectra 2 with AQUA technology provides a reproducible and robust assay which improves section prior to final scoring by impacting overall workflow. AQUA through Vectra Review. of whole tissue section samples generates workflow in a clinical setting. comparable scores by AQUA. Genoptix® is a registered trademark of Genoptix, Inc. © 2013 Genoptix, Inc.Hays_AACR_poster.indd 1 4/3/13 4:52 PM