Color deconvolution for the analysis of tissue microarrays.
OBJECTIVE
To analyze tissue microarrays (TMAs) using color deconvolution, a method for separating component dyes in digital images, and compare the results to observer scoring.


METHODS
TMAs were constructed from tissues from 100 adult autopsies and immunohistochemically stained for connective tissue growth factor. A region of interest (ROI) was created for each core image using 3 binary masks-tissue area, inclusion area and exclusion area. The diaminobenzidine (DAB) and hematoxylin signals were deconvolved, and the DAB signal was measured for each ROI. The dorsalis pedis core images were also scored manually.


RESULTS
Seventeen TMAs were annotated, requiring 532 minutes. Of the 1,683 cores, 296 (18%) were excluded because they were not suitable for evaluation. A single TMA required a mean of 31.3 minutes to evaluate; to annotate a single core took a mean of 19 seconds. For the dorsalis pedis, observer score and median DAB intensity correlated strongly (Kendall's = 0.71).


CONCLUSIONS
Analysis of TMAs by color deconvolution is efficient and highly correlates to observer scoring.
Cornish
Toby C
TC
Department of Pathology, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA. tcornis3@jhmi.edu
Halushka
Marc K
MK
D015100:3,3'-Diaminobenzidine / Q000378:metabolism; D000328:Adult; D015415:Biomarkers / Q000378:metabolism; D001808:Blood Vessels / Q000378:metabolism / Q000473:pathology; D003116:Color; D055513:Connective Tissue Growth Factor / Q000378:metabolism; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D007668:Kidney / Q000378:metabolism / Q000473:pathology; D008168:Lung / Q000378:metabolism / Q000473:pathology; D015203:Reproducibility of Results; D013194:Staining and Labeling / Q000379:methods; D046888:Tissue Array Analysis / Q000379:methods*
D015415:Biomarkers; C526317:CCN2 protein, human; D055513:Connective Tissue Growth Factor; D015100:3,3'-Diaminobenzidine