[2144] Multispectral Cytomorphometric Analysis with Receiver Operating Characteristic Analysis: A Mathematical Approach to Anatomic Pathology Applied to the Study of Barrett Esophagus

Sang Hoon Lee, Emma E Furth. Hospital of the University of Pennsylvania, Philadelphia

Background: In this study, we present our development of a rigorous, quantitative methodology coupled with receiver operating characteristic (ROC) analysis, a mathematical method developed from signal detection theory. We apply this method to the study of epithelium in Barrett esophagus. While nuclear morphology is the feature distinguishing reactive versus dysplastic cells, we hypothesized that quantifiable peri-nuclear characteristics, perhaps reflecting nuclear envelope biology, may also distinguish between these two cell classes.
Design: Cases were taken from surgical pathology esophageal biopsy specimens previously diagnosed by expert pathologists. Fourteen high-grade dysplasia slides and twelve negative-for-dysplasia slides were selected for initial analyses. From each hematoxylin-eosin-stained slide, multiple user-selected image frames at 40x magnification were selected to gather multispectral data in ten nanometer steps from 420 nm to 720 nm. These data cubes were then processed and analyzed against a pure hematoxylin spectrum to ascertain the optical density of hematoxylin staining in the perinuclear cytoplasm for each epithelial cell of interest. The distribution of optical density values for high-grade dysplastic cells was then compared against the distribution for negative cells. Each data set was normalized to 100. ROC analysis was used to determine the area under the curve (AUC) as a metric of this opitcal feature as a test to distinguish these two classes of cell types (AUC=1 being a perfect test, AUC=0.5 being non-descrimitory).
Results: 215 unique high-grade data cubes and 211 negative cubes, each with cell counts ranging from approximately 80-300 cells, were analyzed. The AUC was 0.5 showing no discriminatory value of this feature.


Conclusions: Multispectral cytomorphometry and the application of ROC derived from signal detection theory are powerful tools for anatomic pathology. We conclude that the perinuclear, cytoplasmic, hematoxylin optical density measurements are equivalent in benign and dysplastic epithelial cells in Barret esophagus showing that the biology of the peri-nuclear cytoplasmic environment is independent of the immediately adjacent nuclear envelope.
Category: Techniques

Monday, March 19, 2012 9:30 AM

Poster Session I Stowell-Orbison/Surgical Pathology/Autopsy Awards Poster Session # 314, Monday Morning

 

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