Localized substance delivery to single cell and 4D imaging of its uptake using a flow channel with a lateral aperture
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We have developed a novel microfluidic device for space- and time-resolved (4D) visualization of intracellular events when a cell surface is partially exposed to external stimuli. The device, fabricated using 3D rotational inclined UV lithography of photoresist SU-8, consists of a cell-containing chamber and a flow channel separated by a thin vertical wall having a lateral micrometer aperture smaller than a cell. A cell is first immobilized on the aperture by suction from the flow channel using a syringe pump, and a chemical stimulant is then fed to the channel so that only the cell surface bounded by the aperture is subjected to the stimulus without leakage to other part of the cell surface. The subsequent lateral signal propagation inside the cell can be visualized using high-speed fluorescence confocal microscopy. As an experimental demonstration of the device, 2-NBDG (fluorescence glucose analog) intake into a mouse insulinoma cell, MIN6m9, was visualized in 4D resolution.
KeywordsSingle cell analysis 4D imaging Microfluidic channel Localized substance delivery Multidirectional UV lithography
We thank Prof. S. Seino for providing MIN6m9 cells. This work was supported in part by the research project ‘Development of Bio/Nano Hybrid Platform Technology towards Regenerative Medicine’, CREST, Japan Science and Technology Agency (JST). Part of this work was conducted in the Kyoto-Advanced Nanotechnology Network, supported by the ‘Nanotechnology Network’ of the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan.
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