Single-Cell Biophysics: Measurement, Modulation, and Modeling

Single-Cell Biophysics: Measurement, Modulation, and Modeling

Poster Abstracts

14-POS Board 7 Inspect the Dynamic Variations in Cytoskeleton Arrangement during Cell Aggregating in Time Lapse Microscopy Jiang Ching-Fen . I-Shou University, Kaohsiung, Taiwan. The cytoskeleton is an intricate network of protein filaments that support the cell architecture and dominate cell motility by performing contractility. The structure of cytoskeleton is continuously reorganized as a cell changes shape, divided, and responds to its environment. Therefore, investigation into its dynamic structural variation gives a clue to understand the biological functional change of the cell, such as differentiation, growth, metastasis, and apoptosis. However, current fluorescence microscopic techniques limit to observation on static structure of the cytoskeleton. This study developed a series of image processing techniques to extract the cytoskeleton in the cell images obtained from time-lapse microscopy, and allowed the investigation into the dynamic structure of cytoskeleton during cell migration. Two types of cells, mesenchymal stem cells (MSCs) and lung cancer cells (LCCs), cultured in chitosan-HA membranes were investigated in this study. Microscopic videos were provided by the Biomaterials Laboratory in the Institute of Polymer Science and Engineering at National Taiwan University. The videos were recorded by a ASTEC® CCM-1.4XZY/CO2 system with a CCD camera mounted on a time-lapse microscope with a magnification ratio of 100:1. The acquisition rate was fixed at one acquisition every 15 min. We first verified the consistency between textural patterns extracted by our method and the cytoskeleton distribution viewed by fluorescence microscopy. The dynamic variations in cytoskeleton patterns of the MSCs and LCCs during their aggregation were then compared. The results show that the aggregated LCCs without merging contained concentric textural pattern, while the aggregated MSCs with merging revealed directional texture pattern which corresponded to their moving directions. Hence, the developed cell image processing techniques may provide a direct and convenient way to inspect the dynamic arrangement of cytoskeleton during cell movement without immuno-fluorescence staining

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