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The Effect of Pipette Tip Roughness on Giga-seal Formation

Malboubi, M; Ostadi, H; Wang, S; Gu, Y; Jiang, K; (2009) The Effect of Pipette Tip Roughness on Giga-seal Formation. In: Ao, SI and Gelman, L and Hukins, DWL and Hunter, A and Korsunsky, AM, (eds.) WORLD CONGRESS ON ENGINEERING 2009, VOLS I AND II. (pp. 1849 - 1852). INT ASSOC ENGINEERS-IAENG

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Abstract

Reported is a study of applying nanofabication technology to improve the surface roughness of micro glass pipettes to achieve giga ohm seal resistance in patch clamping processes. The surface roughness of pipette tips was first measured by 3D reconstruction of pipette tips using stereo imaging technique based on high resolution SEM images. Both the SEM images and the reconstructed images show that micro glass pipettes have rough and uneven tips which could be one of the causes of leakage in patch clamping. Then focused ion beam system was used to cut across the very end of the tip, producing a smooth and flat new tip. The average surface roughness Ra of a millet! pipette tip was within a few nanometres. Patch clamping experiments were carried out using the polished pipettes on human umbilical vein endothelial cells (HUVEC), which were well known for their extremely flat shape making them very difficult to patch. The results show that above 3 Giga ohm seals were achieved in 60% of the experiments, as opposed to 1.5-2.0 G Omega in average with the conventional pipettes. The highest seal resistance achieved with a focused ion beam polished pipette was 9 G Omega, well above the 3 G Omega resistance, the usually best result achieved with a conventional pipette. The leakage current in single channel recording afterwards was found 0.3 pA, significantly smaller than 2-3 pA usually achieved using conventional pipettes. The research results demonstrate that the surface roughness of a pipette has a significant effect on the giga-seal formation of a patch clamping process.

Type: Proceedings paper
Title: The Effect of Pipette Tip Roughness on Giga-seal Formation
Event: World Congress on Engineering
Location: Imperial Coll London, London, ENGLAND
Dates: 2009-07-01 - 2009-07-03
ISBN-13: 978-988-17012-5-1
Keywords: Focused ion beam, Giga-seal formation, Patch clamping, Pipette, Roughness, PATCH, MEMBRANE, FABRICATION, CHIP
UCL classification: UCL > Provost and Vice Provost Offices
UCL > Provost and Vice Provost Offices > UCL BEAMS
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Maths and Physical Sciences
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Maths and Physical Sciences > London Centre for Nanotechnology
URI: http://discovery.ucl.ac.uk/id/eprint/1325582
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