Protocol detail

Quantifying Immune Cell Populations in Tumor Tissue

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To evaluate the densities of IL-25⁺, CD4⁺, CD8⁺, CD103⁺ and Foxp3⁺ cells, areas of intra-tumor (IT) and non-tumor (NT) tissue sections were screened by microscopy at low magnification (× 100); five representative spot images were then selected and captured at × 200 magnification using a computerized system that included a Digital Sight DS-Fi1 camera installed on a Nikon Eclipse 80i light microscope (Nikon). Cells that were stained positive were automatically counted using inForm image analysis software (Perkin-Elmer Applied Biosystems; Hopkinton, MA, USA) [48 (link)]. Cell numbers were expressed as mean ± SEM per mm².

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Li J., Liao Y., Ding T., Wang B., Yu X., Chu Y., Xu J, & Zheng L. (2016). Tumor-infiltrating macrophages express interleukin-25 and predict a favorable prognosis in patients with gastric cancer after radical resection. Oncotarget, 7(10), 11083-11093.

Publication 2016

Digital Sight DS-Fi1 camera Nikon Eclipse 80i light microscope inForm image analysis software

Cd103 Cells Il 25 Microscope light Microscopy Sight Tissue Tumor

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Variable analysis

independent variables

Tumor tissue (IT) versus non-tumor tissue (NT)

dependent variables

Density of IL-25+ cells
Density of CD4+ cells
Density of CD8+ cells
Density of CD103+ cells
Density of Foxp3+ cells

control variables

Magnification of microscopy (×100 and ×200)
Use of a computerized system with a Digital Sight DS-Fi1 camera and Nikon Eclipse 80i light microscope
Use of inForm image analysis software for automated cell counting

controls

Positive control: Not specified
Negative control: Not specified

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