Koninklijke Bibliotheek, National Library of the Netherlands
IP1497443754496
Endocytic pathways: combined scanning ion conductance and surface confocal microscopy study
Shevchuk, Andrew I.
Hobson, Phil
Lab, Max J.
Klenerman, David
Krauzewicz, Nina
Korchev, Yuri E.
text
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Pflügers Archiv
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Elektronische Wetenschappelijke Tijdschriften
EWTIJ
10.1007/s00424-007-0410-4
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Springer-Verlag
Berlin/Heidelberg
424
0031-6768
1432-2013
Pflügers Archiv - European Journal of Physiology
European Journal of Physiology
Pflugers Arch - Eur J Physiol
Biomedicine
Human Physiology
456
456
6
1
1
Atomic force microscopy enters physiology
22
2008
3
20
2008
3
20
2008
4
Springer-Verlag
2008
410
10.1007/s00424-007-0410-4
20
Endocytic pathways: combined scanning ion conductance and surface confocal microscopy study
Invited Review
227
235
2007
11
21
2007
9
28
2007
11
20
2008
1
5
The Author(s)
2007
Andrew
I.
Shevchuk
Phil
Hobson
Max
J.
Lab
David
Klenerman
Nina
Krauzewicz
Yuri
E.
Korchev
+44-208-3833080
+44-208-3838306
y.korchev@imperial.ac.uk
grid.7445.2
0000000121138111
MRC Clinical Sciences Centre, Faculty of Medicine
Imperial College London
Hammersmith Hospital Campus, Du Cane Road
London
W12 0NN
UK
grid.13097.3c
0000000123226764
Randall Division of Cell and Molecular Biophysics, New Hunt’s House
King’s College London
Guy’s Campus
London
SE1 1UL
UK
grid.5335.0
0000000121885934
Department of Chemistry
Cambridge University
Cambridge
CB2 1EW
UK
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 ~200-nm indentations in the cell membrane that supports involvement of this protein in endocytosis.
Keywords
Caveola
Endocytosis
Fluorescence
Membrane topology
Membrane transport
A. Shevchuk and P. Hobson contributed equally to this work.
Springer-Verlag
Berlin/Heidelberg
424
0031-6768
1432-2013
Pflügers Archiv - European Journal of Physiology
European Journal of Physiology
Pflugers Arch - Eur J Physiol
Biomedicine
Human Physiology
456
456
6
1
1
Atomic force microscopy enters physiology
22
2008
3
20
2008
3
20
2008
4
Springer-Verlag
2008
410
10.1007/s00424-007-0410-4
20
Endocytic pathways: combined scanning ion conductance and surface confocal microscopy study
Invited Review
227
235
2007
11
21
2007
9
28
2007
11
20
2008
1
5
The Author(s)
2007
Andrew
I.
Shevchuk
Phil
Hobson
Max
J.
Lab
David
Klenerman
Nina
Krauzewicz
Yuri
E.
Korchev
+44-208-3833080
+44-208-3838306
y.korchev@imperial.ac.uk
grid.7445.2
0000000121138111
MRC Clinical Sciences Centre, Faculty of Medicine
Imperial College London
Hammersmith Hospital Campus, Du Cane Road
London
W12 0NN
UK
grid.13097.3c
0000000123226764
Randall Division of Cell and Molecular Biophysics, New Hunt’s House
King’s College London
Guy’s Campus
London
SE1 1UL
UK
grid.5335.0
0000000121885934
Department of Chemistry
Cambridge University
Cambridge
CB2 1EW
UK
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 ~200-nm indentations in the cell membrane that supports involvement of this protein in endocytosis.
Keywords
Caveola
Endocytosis
Fluorescence
Membrane topology
Membrane transport
A. Shevchuk and P. Hobson contributed equally to this work.
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