Mp27-15 a comparison of computer-assisted analyses and manual counting in vestibular tissue stained with pgp9.5 and cd117

You have accessJournal of UrologyCME1 Apr 2023MP27-15 A COMPARISON OF COMPUTER-ASSISTED ANALYSES AND MANUAL COUNTING IN VESTIBULAR TISSUE STAINED WITH PGP9.5 AND CD117 Alexandra Drian, Noel N. Kim, Sue W. Goldstein, Andrew S Goldstein, Alyssa Yee, and Irwin Goldstein Alexandra DrianAlexandra Drian More articles by this author , Noel N. KimNoel N. Kim More articles by this author , Sue W. GoldsteinSue W. Goldstein More articles by this author , Andrew S GoldsteinAndrew S Goldstein More articles by this author , Alyssa YeeAlyssa Yee More articles by this author , and Irwin GoldsteinIrwin Goldstein More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003255.15AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Immunohistochemical (IHC) analysis of stained sections of vestibular specimens have been used to define diagnostic criteria for neuroproliferative vestibulodynia (NPV). Bornstein’s criteria (2004) included manually counting 8 or more mast cells in a standardized microscopic field and/or subepithelial nerve fibers that were 10 times higher than controls. However, there is wide variability in manual counting, IHC staining and magnification. We examined a more reproducible methodology by assessing percentage of immunopositive stained area using computer-assisted histometry. METHODS: Samples from 17 patients were stained with CD117, a marker of mast cells, and PGP9.5, a neuronal marker. Photomicrographs were obtained using 200x and 400x magnification and analyzed using Image J with 3 measurements per photomicrograph. This methodology was also applied to previously published images of tissues from patients with and without NPV (Tympanidis 2003; Goetsch 2010). For comparison, CD117-immunopositive cells were also manually counted, as described by Bornstein. Clusters of immunopositive granules positively stained and separated from an adjacent cell membrane were counted as a single immunopositive cell. RESULTS: Using Image J, mean CD117-immunopositive area was 2107 ± 1339 μm2, representing 0.79 ± 0.51% of total area analyzed. Mean PGP9.5-immunopositive area was 1123 ± 695 μm2, representing 0.38 ± 0.27% of total area analyzed. Using manual counting, we observed 30.6 ± 15.3 CD117-immunopositive cells per standardized field (200x), greater than the Bornstein threshold of 8 mast cells per standardized field. Tissues from control patients without vestibulodynia had 6 to 8-fold lower immunostained area than tissues from vestibulodynia patients using computer-assisted histometry. In analyses of photomicrographs from Goetsch and Tympanidis, 2.48% and 2.87% of the control areas were immunopositive. For controls, using Image J, immunostained area for PGP 9.5 was 2.6-fold greater than that reported by Bornstein using manual counting (425 μm2). CONCLUSIONS: All our patients had increased density of CD117-immunopositive cells and increased PGP9.5-immunopositive area, confirming their NPV diagnosis. To facilitate more consistent, widespread and rapid assessment of IHC analysis of stained sections of vestibular specimens, we propose using computer-assisted histometry for future studies. Source of Funding: None © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e368 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Alexandra Drian More articles by this author Noel N. Kim More articles by this author Sue W. Goldstein More articles by this author Andrew S Goldstein More articles by this author Alyssa Yee More articles by this author Irwin Goldstein More articles by this author Expand All Advertisement PDF downloadLoading ...