A poly(dimethylsiloxane)-based device enabling time-lapse imaging with high spatial resolution.
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 CO(2)-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.
Hirano
Masahiko
M
Japan Science and Technology Agency, Wako, Saitama 351-0198, Japan. mhirano@postman.riken.jp
Hoshida
Tetsushi
T
Sakaue-Sawano
Asako
A
Miyawaki
Atsushi
A
D002453:Cell Cycle; D004129:Dimethylpolysiloxanes / Q000737:chemistry*; D006367:HeLa Cells; D006801:Humans; D008856:Microscopy, Fluorescence / Q000295:instrumentation* / Q000379:methods*
D004129:Dimethylpolysiloxanes; C013830:baysilon