title: Image analysis for accurately counting CD4+ and CD8+ T cells in human tissue.
abstract: In situ detection of specific cells offers a unique perspective on the spatial interactions between host immune cells and specific viral pathogens or cancers. Most immunohistochemistry techniques require manual cell counting on biopsied and fixed tissue sections. The availability of sophisticated software packages for analyzing fluorescently labeled tissue has made it possible to quickly and accurately quantitate the number of positive cells on such slides. Manual cell counting was compared to automatic cell counting using the program CellProfiler. The two techniques were used to count CD4+ and CD8+ T cells in human genital skin biopsies from herpesvirus type 2 (HSV-2) infected subjects. Manual counting and CellProfiler demonstrated high correlation both in cell counting as well as detection of immune cell "clustering" in tissue, an important visceral component of localized inflammation and characteristic of most chronic infections. Overall, CellProfiler is an effective and accurate method in addition to or replacement of manual cell counting of fluorescently labeled biopsies.
authors.affiliation: Department of Laboratory Medicine, RR-512 Health Sciences Building, University of Washington, Box 356420, 1959 NE Pacific Street, Seattle, WA 98195, USA. Electronic address: diem@u.washington.edu.
authors.forename: Kurt
authors.initials: K
authors.lastname: Diem
authors.affiliation: Department of Laboratory Medicine, RR-512 Health Sciences Building, University of Washington, Box 356420, 1959 NE Pacific Street, Seattle, WA 98195, USA; Vaccine and Infectious Disease Division, Fred Hutchison Cancer Research Center, 1100 Fairview Avenue N, PO Box 19024, Seattle, WA 98109, USA.
authors.forename: Amalia
authors.initials: A
authors.lastname: Magaret
authors.affiliation: Department of Laboratory Medicine, RR-512 Health Sciences Building, University of Washington, Box 356420, 1959 NE Pacific Street, Seattle, WA 98195, USA.
authors.forename: Alexis
authors.initials: A
authors.lastname: Klock
authors.affiliation: Department of Laboratory Medicine, RR-512 Health Sciences Building, University of Washington, Box 356420, 1959 NE Pacific Street, Seattle, WA 98195, USA.
authors.forename: Lei
authors.initials: L
authors.lastname: Jin
authors.affiliation: Department of Laboratory Medicine, RR-512 Health Sciences Building, University of Washington, Box 356420, 1959 NE Pacific Street, Seattle, WA 98195, USA; Vaccine and Infectious Disease Division, Fred Hutchison Cancer Research Center, 1100 Fairview Avenue N, PO Box 19024, Seattle, WA 98109, USA.
authors.forename: Jia
authors.initials: J
authors.lastname: Zhu
authors.affiliation: Department of Laboratory Medicine, RR-512 Health Sciences Building, University of Washington, Box 356420, 1959 NE Pacific Street, Seattle, WA 98195, USA; Vaccine and Infectious Disease Division, Fred Hutchison Cancer Research Center, 1100 Fairview Avenue N, PO Box 19024, Seattle, WA 98109, USA.
authors.forename: Lawrence
authors.initials: L
authors.lastname: Corey
mesh_terms: D001706:Biopsy; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D003584:Cytological Techniques / Q000379:methods*; D005453:Fluorescence; D006558:Herpes Genitalis / Q000473:pathology; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D018655:Lymphocyte Count / Q000379:methods*; D010336:Pathology / Q000379:methods*; D012867:Skin / Q000473:pathology; D013194:Staining and Labeling
keywords: Cell counting; CellProfiler; HSV-2; Immunohistochemistry (IHC); Software cell counting