Automated In-situ Analysis of Tumor-Associated Macrophage Attachment on Antifouling Polymer Brushes

Abstract

Tumor-associated macrophages (TAMs) are critical to tumor progression. Quantifying their interactions with biomaterial surfaces is crucial for developing effective cancer therapies. Traditionally, manual cell counting has been used to assess macrophage adhesion, a labor-intensive and subjective process. To address these limitations and enable unbiased analysis, we developed an automated in-situ system to quantify TAM attachment to antifouling polymer brushes. Bland-Altman analysis indicated a high agreement between our automated method and traditional manual cell counting. For M1 macrophages, the mean difference was less than 4 cells, with limits of agreement (LoA) ranging from -70.18% to 80.16%. For M2 macrophages, the mean difference was 25 cells, with LoA ranging from -51.61% to 72.71%. These results were consistent across different experimental conditions, including Unspecific Binding, Specific Antibody, and IgG Control. Our analysis revealed no systematic differences in cell counts and holds significant potential for point-of-care applications, potentially enhancing personalized treatment strategies.