Guideline for in vivo assessment of adherent and rolling leukocytes in human skin microvasculature via reflectance confocal videomicroscopy.

Publisher: Wiley-Blackwell Country of Publication: United States NLM ID: 9434935 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1549-8719 (Electronic) Linking ISSN: 10739688 NLM ISO Abbreviation: Microcirculation Subsets: MEDLINE

Publication: Malden, MA : Wiley-Blackwell
Original Publication: New York, NY : Chapman & Hall, c1994-

Leukocytes* ; Microvessels*/diagnostic imaging; Cell Adhesion ; Humans ; Microcirculation ; Microscopy, Video ; Reproducibility of Results

Objective: To develop a guideline that reliably identifies cutaneous adherent and rolling leukocytes from mimicking scenarios via in vivo reflectance confocal videomicroscopy.
Methods: We used a clinical reflectance confocal microscope, the VivaScope 1500, to acquire 1522 videos of the upper dermal microcirculation from 12 healthy subjects and 60 patients after allogeneic hematopoietic cell transplantation. Blinded to clinical information, two trained raters independently counted the number of adherent and rolling leukocytes in 88 videos. Based on discrepancies in the initial assessments, we developed a guideline to identify both types of leukocyte-endothelial interactions via a modified Delphi method (without anonymity). To test the guideline's ability to improve the inter-rater reliability, the two raters assessed the remaining 1434 videos by using the guideline.
Results: We demonstrate a guideline that consists of definitions, a step-by-step flowchart, and corresponding visuals of adherent and rolling leukocytes and mimicking scenarios. The guideline improved the inter-rater reliability of the manual assessment of both interactions. The intraclass correlation coefficient (ICC) of adherent leukocyte counts increased from 0.056 (95% confidence interval: 0-0.236, n = 88 videos, N = 10 subjects) to 0.791 (0.770-0.809, n = 1434, N = 67). The ICC of rolling leukocyte counts increased from 0.385 (0.191-0.550, n = 88, N = 10) to 0.626 (0.593-0.657, n = 1434, N = 67). Intra-rater ICC post-guideline was 0.953 (0.886-0.981, n = 20, N = 12) and 0.956 (0.894-0.983, n = 20, N = 12) for adherent and rolling, respectively.
Conclusion: The guideline aids in the manual identification of adherent and rolling leukocytes via in vivo reflectance confocal videomicroscopy.
(© 2021 John Wiley & Sons Ltd. This article has been contributed to by US Government employees and their work is in the public domain in the USA.)

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IK2 CX001785 United States CX CSRD VA; K12 CA090625 United States CA NCI NIH HHS

Keywords: adhesion; confocal; leukocyte; noninvasive; rolling; videomicroscopy

Date Created: 20210819 Date Completed: 20220324 Latest Revision: 20221102

20221213

PMC8602735

10.1111/micc.12725

34409720