A poly(dimethylsiloxane)-based device enabling time-lapse imaging with high spatial resolution |
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Authors: | Masahiko Hirano Asako Sakaue-Sawano |
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Affiliation: | ERATO Life Function Dynamics Project, Japan Science and Technology Agency, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan Laboratory for Cell Function Dynamics, Advanced Technology Development Group, Brain Science Institute, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan |
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Abstract: | We have developed a regulator-free device that enables long-term incubation of mammalian cells for epi-fluorescence imaging, based on a concept that the size of sample to be gassed and heated is reduced to observation scale. A poly(dimethylsiloxane) block stamped on a coverslip works as a long-lasting supplier of CO2-rich gas to adjust bicarbonate-containing medium in a tiny chamber at physiological pH, and an oil-immersion objective warms cells across the coverslip. A time-lapse imaging experiment using HeLa cells stably expressing fluorescent cell-cycle indicators showed that the cells in the chamber proliferated with normal cell-cycle period over 2 days. |
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Keywords: | Time-lapse imaging Fluorescence imaging Cell cycle Poly(dimethylsiloxane) Bicarbonate-containing medium |
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