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Cellular response to physical stretch has been extensively studied as a regulator of various physiological functions. For live cell microscopy combined with stretch experiments, cells are typically seeded on an extensible elastomer sheet. In this case, the position of the cells of interest tends to shift out of the field of view upon stretch, making real-time imaging of identical cells difficult. To circumvent this situation, here we describe a robust methodology in which these cell shifts are minimized. Cells are plated in a custom-designed stretch chamber with an elastomer sheet of a small cell culture area. The cell-supporting chamber is stretched on an inverted microscope by using a piezoelectric actuator that provides small, but precisely controlled displacements. Even under this small displacement within the filed of view, our device allows the cells to undergo physiologically relevant levels of stretch. Identical cells can thus be continuously observed during stretching, thereby potentially enabling imaging of stretch-triggered fast dynamics.


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