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Image analysis for accurately counting CD4+ and CD8+ T cells in human tissue
Kurt Diem1, Amalia Magaret2, Alexis Klock1
1Department of Laboratory Medicine, RR-512 Health Sciences Building, University of Washington, Box 356420, 1959 NE Pacific Street, Seattle, WA 98195, USA.
Insights
Automated cell counting using CellProfiler accurately quantifies immune cells in herpesvirus type 2 (HSV-2) infected biopsies. This method correlates well with manual counting and detects immune cell clustering, offering an efficient alternative for analyzing fluorescently labeled tissues.
Area of Science:
- Immunology
- Virology
- Computational Biology
Background:
- In situ detection of immune cells provides insights into host-pathogen interactions.
- Traditional manual cell counting in tissue sections is time-consuming and subjective.
- Automated image analysis offers potential for faster and more accurate quantification.
Purpose of the Study:
- To compare manual cell counting with automated cell counting using CellProfiler.
- To evaluate the accuracy and efficiency of CellProfiler for analyzing immune cells in infected tissues.
- To assess the ability of CellProfiler to detect immune cell clustering.
Main Methods:
- Fluorescently labeled human genital skin biopsies from herpesvirus type 2 (HSV-2) infected subjects were analyzed.
- Manual cell counting was performed on tissue sections.
- Automated cell counting and clustering analysis were conducted using CellProfiler software.
Main Results:
- CellProfiler demonstrated high correlation with manual cell counting for both CD4+ and CD8+ T cells.
- Automated analysis accurately detected immune cell clustering, a key indicator of inflammation.
- CellProfiler provides a rapid and precise method for quantifying immune cells in situ.
Conclusions:
- CellProfiler is an effective and accurate tool for quantifying immune cells in fluorescently labeled biopsies.
- Automated analysis can supplement or replace manual counting in research and diagnostics.
- This approach enhances the study of immune responses in infectious diseases and other conditions.
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.
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