Endocytic pathways: combined scanning ion conductance and surface confocal microscopy study

Pflügers Archiv : European journal of physiology
Andrew I ShevchukYuri E Korchev

Abstract

We introduce a novel high resolution scanning surface confocal microscopy technique that enables imaging of endocytic pits in apical membranes of live cells for the first time. The improved topographical resolution of the microscope together with simultaneous fluorescence confocal detection produces pairs of images of cell surfaces sufficient to identify single endocytic pits. Whilst the precise position and size of the pit is detected by the ion conductance microscope, the molecular nature of the pit, e.g. clathrin coated or caveolae, is determined by the corresponding green fluorescent protein fluorescence. Also, for the first time, we showed that flotillin 1 and 2 can be found co-localising with approximately 200-nm indentations in the cell membrane that supports involvement of this protein in endocytosis.

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Citations

Dec 9, 2008·Antioxidants & Redox Signaling·Hemal H Patel, Paul A Insel
Dec 26, 2012·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·David KlenermanSimon J Davis
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May 9, 2012·The Journal of Cell Biology·Andrew I ShevchukYuri E Korchev
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Apr 25, 2019·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Tayyibah AliAndrew Shevchuk
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Jul 20, 2012·Physiological Reviews·Elise BalseStéphane N Hatem
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Jan 19, 2018·Analytical Chemistry·Yuanshu ZhouYasufumi Takahashi
Dec 10, 2020·Chemical Reviews·Cheng ZhuLane A Baker

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Methods Mentioned

BETA
confocal microscopy
fluorescence-based microscopy
electron microscopy
FCS
transfection

Software Mentioned

SICM

Related Concepts

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