Holographic Cell Stiffness Mapping Using Acoustic Stimulation

Abstract

Accurate assessment of stiffness distribution is essential due to the critical role of single cell mechanobiology in the regulation of many vital cellular processes such as proliferation, adhesion, migration, and motility. Cell stiffness is one of the fundamental mechanical properties of the cell and is greatly affected by the intracellular tensional forces, cytoskeletal prestress, and cytoskeleton structure. Herein, we propose a novel holographic single-cell stiffness measurement technique that can obtain the stiffness distribution over a cell membrane at high resolution and in real-time. The proposed imaging method coupled with acoustic signals allows us to assess the cell stiffness distribution with a low error margin and label-free manner. We demonstrate the proposed technique on HCT116 (Human Colorectal Carcinoma) cells and CTC-mimicked HCT116 cells by induction with transforming growth factor-beta (TGF-\b{eta}). Validation studies of the proposed approach were carried out on certified polystyrene microbeads with known stiffness levels. Its performance was evaluated in comparison with the AFM results obtained for the relevant cells. When the experimental results were examined, the proposed methodology shows utmost performance over average cell stiffness values for HCT116, and CTC-mimicked HCT116 cells were found as 1.08 kPa, and 0.88 kPa, respectively. The results confirm that CTC-mimicked HCT116 cells lose their adhesion ability to enter the vascular circulation and metastasize. They also exhibit a softer stiffness profile compared to adherent forms of the cancer cells. Hence, the proposed technique is a significant, reliable, and faster alternative for in-vitro cell stiffness characterization tools. It can be utilized for various applications where single-cell analysis is required, such as disease modeling, drug testing, diagnostics, and many more.