Glass slide HPF at × 400 magnification with conventional light microscopy was not equivalent to '×40' digital HPF areas. No differences were identified by scanner or WSIs scanned at different resolutions (e.g., 0.5, 0.25, 0.24, and 0.12 μm/pixel). Digital and microscopic HPF areas were calculated and compared.Ī significant variation of HPF area occurred between differing monitor size and display resolutions with minor differences between WSI viewers. WSIs were reviewed at '×40' equivalent HPF on different sized monitors with varying display resolutions (1900 × 1080-4500 × 3000) and aspect ratios (e.g., Food and Drug Administration -cleared 27' Philips PS27QHDCR, FDA-cleared 24' Dell MR2416, 24' Hewlett Packard Z24n G2, and 28' Microsoft Surface Studio). Glass slides scanned on various devices (Leica's Aperio GT450, AT2, and ScanScope XT Philips UltraFast Scanner Hamamatsu's Nanozoomer 2.0HT and 3DHistech's P1000) were compared to acquired digital slides reviewed on each vendor's respective WSI viewer software (e.g., Aperio ImageScope, ImageScope DX, Philips IMS, 3DHistech CaseViewer, and Hamamatsu NDP.view) and an in-house developed vendor-agnostic viewer. The aim of this study was to determine the parameters that influence HPF in whole slide images (WSIs). In the era of digital pathology, new variables are introduced that may affect HPF evaluation.

With traditional light microscopy, HPFs are typically evaluated by quantifying histologic events in 10 fields of view at × 400 magnification. The microscope high-power field (HPF) is the cornerstone for histopathology diagnostic evaluation such as the quantification of mitotic figures, lymphocytes, and tumor grading.

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