Ion channels in small cells and subcellular structures can be studied with a smart patch-clamp system.
We have developed a scanning patch-clamp technique that facilitates single-channel recording from small cells and submicron cellular structures that are inaccessible by conventional methods. The scanning patch-clamp technique combines scanning ion conductance microscopy and patch-clamp recording through a single glass nanopipette probe. In this method the nanopipette is first scanned over a cell surface, using current feedback, to obtain a high-resolution topographic image. This same pipette is then used to make the patch-clamp recording. Because image information is obtained via the patch electrode it can be used to position the pipette onto a cell with nanometer precision. The utility of this technique is demonstrated by obtaining ion channel recordings from the top of epithelial microvilli and openings of cardiomyocyte T-tubules. Furthermore, for the first time we have demonstrated that it is possible to record ion channels from very small cells, such as sperm cells, under physiological conditions as well as record from cellular microstructures such as submicron neuronal processes.
Gorelik
Julia
J
Division of Medicine, Imperial College of Science, Technology and Medicine, MRC Clinical Sciences Centre, DuCane Road, London W12 0NN, United Kingdom.
Gu
Yuchun
Y
Spohr
Hilmar A
HA
Shevchuk
Andrew I
AI
Lab
Max J
MJ
Harding
Sian E
SE
Edwards
Christopher R W
CR
Whitaker
Michael
M
Moss
Guy W J
GW
Benton
David C H
DC
Sánchez
Daniel
D
Darszon
Alberto
A
Vodyanoy
Igor
I
Klenerman
David
D
Korchev
Yuri E
YE
D000818:Animals; D001011:Aorta / Q000502:physiology / Q000648:ultrastructure; D002460:Cell Line; D004847:Epithelial Cells / Q000502:physiology / Q000648:ultrastructure; D004867:Equipment Design; D005240:Feasibility Studies; D005246:Feedback; D066298:In Vitro Techniques; D007425:Intracellular Membranes / Q000502:physiology / Q000648:ultrastructure; D007473:Ion Channels / Q000502:physiology* / Q000648:ultrastructure; D007668:Kidney / Q000502:physiology / Q000648:ultrastructure; D008297:Male; D008564:Membrane Potentials / Q000502:physiology; D008566:Membranes / Q000502:physiology / Q000648:ultrastructure; D020527:Microscopy, Scanning Probe / Q000295:instrumentation* / Q000379:methods*; D032383:Myocytes, Cardiac / Q000502:physiology / Q000648:ultrastructure; D009474:Neurons / Q000502:physiology / Q000648:ultrastructure; D018408:Patch-Clamp Techniques / Q000295:instrumentation* / Q000379:methods*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012617:Sea Urchins / Q000502:physiology / Q000648:ultrastructure; D013094:Spermatozoa / Q000502:physiology / Q000648:ultrastructure
D007473:Ion Channels