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Validating Whole Slide Digital Morphometric Analysis as a Microscopy Tool

  • Robert B. Diller (a1) and Robert S. Kellar (a1) (a2)


Whole slide imaging (WSI) can be used to quantify multiple responses within tissue sections during histological analysis. Feature Analysis on Consecutive Tissue Sections (FACTS®) allows the investigator to perform digital morphometric analysis (DMA) within specified regions of interest (ROI) across multiple serial sections at faster rates when compared with manual morphometry methods. Using FACTS® in conjunction with WSI is a powerful analysis tool, which allows DMA to target specific ROI across multiple tissue sections stained for different biomarkers. DMA may serve as an appropriate alternative to classic, manual, histologic morphometric measures, which have historically relied on the selection of high-powered fields of views and manual scoring (e.g., a gold standard). In the current study, existing preserved samples were used to determine if DMA would provide similar results to manual counting methods. Rodent hearts (n=14, left ventricles) were stained with Masson’s trichrome, and reacted for cluster of differentiation 68 (CD-68). This study found no statistical significant difference between a classic, manual method and the use of digital algorithms to perform the similar counts (p=0.38). DMA offers researchers the ability to accurately evaluate morphological characteristics in a reproducible fashion without investigator bias and with higher throughput.


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Al-Janabi, S., Huisman, A. & Van Diest, P.J. (2012). Digital pathology: Currents status and future perspectives. Histopathology 61, 19.
Cole, B., Gomoll, A., Yanke, A., Pylawka, T., Lewis, P., MacGillivray, J. & Williams, J. (2007). Biocompatibility of a polymer patch for rotator cuff repair. Knee Surg Sports Traumatol Arthrosc 15(5), 632637.
Crowley, R.S., Gadd, C.S., Naus, G., Becich, M. & Lowe, H.J. (2000). Defining the role of anatomic pathology images in the multimedia electronic medical record—a preliminary report. Proc AMIA Symp, 161165.
Daniel, C., Rojo, M.G., Klossa, J., Mea, V.D., Booker, D., Beckwith, B.A. & Schrader, T. (2011). Standardizing the use of whole slide images in digital pathology. Comput Med Imaging Graph 35(7–8), 496505.
Doussis, I.A., Gatter, K.C. & Mason, D.Y. (1993). CD68 reactivity of non-macrophage derived tumours in cytological specimens. Journal of clinical pathology 46(4), 334336.
Evans, A.J., Henry, P.C., Van der Kwast, T.H., Tkachuk, D.C., Watson, K., Lockwood, G.A. & Srigley, J.R. (2008). Interobserver variability between expert urologic pathologists for extraprostatic extension and surgical margin status in radical prostatectomy specimens. Am J Surg Pathol 32(10), 15031512.
Feldman, M.D. (2008). Beyond morphology: Whole slide imaging, computer-aided detection, and other techniques. Arch Pathol Lab Med 132(5), 758763.
Fujimoto, K.L., Tobita, K., Merryman, W.D., Guan, J., Momoi, N., Stolz, D.B. & Wagner, W.R. (2007). An elastic, biodegradable cardiac patch induces contractile smooth muscle and improves cardiac remodeling and function in subacute myocardial infarction. J Am Coll Cardiol 49(23), 22922300.
Hipp, J., Flotte, T., Monaco, J., Cheng, J., Madabhushi, A., Yagi, Y., Rodriguez-Canales, J., Emmert-Buck, M., Dugan, M., Hewitt, S., Toner, M., Tompkins, R., Lucas, D., Gilbertson, J. & Balis, U. (2011). Computer aided diagnostic tools aim to empower rather than replace pathologists: Lessons learned from computational chess. J Pathol Inform 2, 25.
Hsu, C.Y., Ho, D.M.T., Yang, C.F. & Chiang, H. (2003). Interobserver reproducibility of MIB-1 labeling index in astrocytic tumors using different counting methods. Mod Pathol 16(9), 951957.
Huisman, A., Looijen, A., van den Brink, S.M. & van Diest, P.J. (2010). Creation of a fully digital pathology slide archive by high-volume tissue slide scanning. Hum Pathol 41(5), 751757.
Jara-Lazaro, A.R., Thamboo, T.P., Teh, M. & Tan, P.H. (2010). Digital pathology: Exploring its applications in diagnostic surgical pathology practice. Pathology 42(6), 512518.
Kellar, R.S., Lancaster, J.J., Thai, H.M., Juneman, E., Johnson, N.M., Byrne, H.G., Stansifer, M., Arsanjani, R., Baer, M., Bebbington, C., Flashner, M., Yarranton, G. & Goldman, S. (2011). Antibody to granulocyte macrophage colony-stimulating factor reduces the number of activated tissue macrophages and improves left ventricular function after myocardial infarction in a rat coronary artery ligation model. Journal of cardiovascular pharmacology 57(5), 568574.
Kidd, K.R., Dal Ponte, D.B., Kellar, R.S. & Williams, S.K. (2001). A comparative evaluation of the tissue responses associated with polymeric implants in the rat and mouse. J Biomed Mater Res 59(4), 682689.
Klapczynski, M., Gagne, G.D., Morgan, S.J., Larson, K.J., LeRoy, B.E., Blomme, E.A. & Shek, E.W. (2011). Computer-assisted imaging algorithms facilitate histomorphometric quantification of kidney damage in rodent renal failure models. J Pathol Inform 3, 20.
Lofgren, K.A., Ostrander, J.H., Housa, D., Hubbard, G.K., Locatelli, A., Bliss, R.L. & Lange, C.A. (2011). Mammary gland specific expression of Brk/PTK6 promotes delayed involution and tumor formation associated with activation of p38 MAPK. Breast Cancer Res 13(5), R89.
López, C., Lejeune, M., Salvadó, M.T., Escrivà, P., Bosch, R., Pons, L.E. & Jaén, J. (2008). Automated quantification of nuclear immunohistochemical markers with different complexity. Histochem Cell Biol 129(3), 379387.
Mulrane, L., Rexhepaj, E., Penney, S., Callanan, J.J. & Gallagher, W.M. (2008). Automated image analysis in histopathology: a valuable tool in medical diagnostics. Expert Reviews of Molecular Diagostics 8.6: 707.
Nassar, A., Cohen, C., Albitar, M., Agersborg, S.S., Zhou, W., Lynch, K.A. & Siddiqui, M.T. (2011). Reading immunohistochemical slides on a computer monitor—a multisite performance study using 180 HER2-stained breast carcinomas. Appl Immunohistochem Mol Morphol 19(3), 212217.
Potts, S.J., Johnson, T.D., Voelker, F.A., Lange, H. & Young, G.D. (2011). Multiplexed measurement of proteins in tissue in a clinical environment. Appl Immunohistochem Mol Morphol 19(6), 494498.
Potts, S.J., Young, G.D. & Voelker, F.A. (2010). The role and impact of quantitative discovery pathology. Drug Discov Today 15(21–22), 943950.
Rocha, R., Vassallo, J., Soares, F., Miller, K. & Gobbi, H. (2009). Digital slides: Present status of a tool for consultation, teaching, and quality control in pathology. Pathol Res Pract 205(11), 735741.
Saraswati, S., Alfaro, M.P., Thorne, C.A., Atkinson, J., Lee, E., & Young, P.P. (2010). Pyrvinium, a potent small molecule Wnt inhibitor, promotes wound repair and post-MI cardiac remodeling. PLoS One 5(11), e15521.
Segura, A.M., Frazier, O.H., Demirozu, Z. & Buja, L.M. (2011). Histopathologic correlates of myocardial improvement in patients supported by a left ventricular assist device. Cardiovascular Pathology 20(3), 139145.
Słodkowska, J., Filas, V., Buszkiewicz, E., Trzeciak, P., Wojciechowski, M., Koktysz, R. & Garcia Rojo, M. (2010). Study on breast carcinoma Her2/neu and hormonal receptors status assessed by automated images analysis systems: ACIS III (Dako) and ScanScope (Aperio). Folia Histochem Cytobiol 48(1), 1925.
Wang, Y., Turner, R., Crookes, D., Diamond, J. & Hamilton, P. (2007). Investigation of methodologies for the segmentation of squamous epithelium from cervical histological virtual slides. In Machine Vision and Image Processing Conference, 2007 IMVIP 2007, pp. 83–90. IEEE, NUI Maynooth, Ireland.
Webster, J.D. & Dunstan, R.W. (2014). Whole-slide imaging and automated image analysis considerations and opportunities in the practice of pathology. Vet Pathol Online 51(1), 211223.
Weinstein, R.S., Graham, A.R., Richter, L.C., Barker, G.P., Krupinski, E.A., Lopez, A.M. & Gilbertson, J.R. (2009). Overview of telepathology, virtual microscopy, and whole slide imaging: Prospects for the future. Hum Pathol 40(8), 10571069.


Validating Whole Slide Digital Morphometric Analysis as a Microscopy Tool

  • Robert B. Diller (a1) and Robert S. Kellar (a1) (a2)


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