Title: Multimodal Analysis of Traction Forces and Temperature Dynamics of Living Cells with Diamond-Embedded Substrate

Abstract: Cells and tissues are constantly exposed to various chemical and physical signals that intricately regulate various physiological and pathological processes. This study explores the integration of two biophysical methods, Traction Force Microscopy (TFM) and Optically-Detected Magnetic Resonance (ODMR), to concurrently assess cellular traction forces and local relative temperature. We present a novel elastic substrate with embedded nitrogen-vacancy microdiamonds, that facilitate ODMR-TFM measurements. Optimization efforts have focused on minimizing the sample illumination and experiment duration to mitigate biological perturbations. Our hybrid ODMR-TFM technique yields precise TFM maps and achieves approximately 1K accuracy in relative temperature measurements. Notably, our setup, employing a simple wide-field fluorescence microscope with standard components, demonstrates the broader feasibility of these techniques in life-science laboratories. By elucidating the physical aspects of cellular behavior beyond the existing methods, this approach opens avenues for a deeper understanding and may inspire diverse biomedical applications.