Journal
LAB ON A CHIP
Volume 11, Issue 18, Pages 3064-3071Publisher
ROYAL SOC CHEMISTRY
DOI: 10.1039/c1lc20458k
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Funding
- Austrian Science Fund [W1201-N13]
- (Doktoratskolleg: Molekulare Bioanalytik) be the Government of Upper Austria [Wi(Ge)-213970-3]
- (Hochsensitive Analysemethoden fur die Untersuchung von biologischen Proben) by the European Fund for Regional Development [Wi-219244/20-2010/Kr/Zs]
- Government of Upper Austria
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A poly(dimethylsiloxane) (PDMS)-based biochip with an integrated pressure controlled positioning system with sub-micrometre precision was realized. The biochip was easy and cheap to manufacture and enabled positioning in a wet environment. It allowed the application of total internal reflection fluorescence (TIRF) microscopy at the dorsal cell membrane, which is not adhering to a support. Specifically, the chip enabled TIRF microscopy at the apical membrane of polarized epithelial cells. Thereby, the device allowed us for the first time to monitor individual fusion events of GPI-GFP bearing vesicles at the apical membrane in live Madin-Darby canine kidney II (MDCK II) cells. Moreover, a mapping of fusion sites became feasible and revealed that the whole apical membrane is fusion competent. In total, the biochip offers an all-in-one solution for apical TIRF microscopy and contributes a novel tool to study trafficking processes close to the apical plasma membrane in polarized epithelial cells.
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