Work

Longitudinal, label-free, high-resolution imaging of glioblastoma spheroid response to therapy: a translational tool for preclinical evaluation of chemotherapy, radiation, and immunotherapy

Year: 2025

Type: article

Abstract: platform for evaluating treatments such as chimeric antigen receptor (CAR) T cell therapy, chemotherapy, and radiation. However, significant challenges remain in monitoring the complex three-dimensional (3D) microenvironment of the GBM PDTSs. Current imaging techniques used for this purpose are primarily endpoint analyses which lack critical real-time, non-invasive capabilities that ultimately preclude longitudinal and continuous monitoring. In this study, we introduce quantitative oblique back-illumination microscopy (qOBM) as a label-free and non-invasive imaging approach for longitudinal and continuous, high-resolution monitoring of GBM PDTSs during treatment. qOBM enables real-time visualization of cellular processes, including apoptosis, cell migration, and T cell-mediated cytotoxicity by leveraging tomographic refractive index-based quantitative imaging. We construct a compact qOBM system that fits within common incubators and apply it to study the effects of radiation, chemotherapy, and immunotherapy on three patient-derived GBM cell lines, extracting both static and dynamic image features over a 72 h treatment period. Additionally, we develop machine learning models to predict spheroid viability and cytotoxicity, demonstrating the potential of qOBM to enhance treatment evaluation. Our findings establish qOBM as a powerful tool for longitudinal and continuous spheroid monitoring, offering a non-destructive, high-resolution alternative to conventional endpoint assays and improving the evaluation of preclinical treatments for GBM.

Source: Optica

DOI: 10.1364/optica.574860