Patch clamping on plane glass-fabrication of hourglass aperture and high-yield ion channel recording.
Planar patch-clamp has revolutionized ion-channel measurement by eliminating laborious manipulation from the traditional micropipette approach and enabling high throughput. However, low yield in gigaseal formation and/or relatively high cost due to microfabricated processes are two main drawbacks. This paper presents patch clamping on glass substrate-an economical solution without sacrificing gigaseal yield rate. Two-stage CO(2) laser drilling methodology was used to generate an hourglass, funnel-like aperture of a specified diameter with smooth and debris-free surfaces on 150 microm borosilicate cover glass. For 1-3 microm apertures as patch-clamp chips, seal resistance was tested on human embryonic kidney, Chinese hamster ovary, and Jurkat T lymphoma cells with a gigaseal success rate of 62.5%, 43.6% and 66.7% respectively. Results also demonstrated both whole-cell and single channel recording on endogenously expressed ion channels to confirm the capability of different patch configurations.
Chen
Chang-Yu
CY
Institute of Applied Mechanics, National Taiwan University, 1 Roosevelt Road, Section 4, Taipei, 106, Taiwan.
Tu
Ting-Yuan
TY
Chen
Chang-Hung
CH
Jong
De-Shien
DS
Wo
Andrew M
AM
D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D004129:Dimethylpolysiloxanes; D004553:Electric Conductivity; D005898:Glass; D006801:Humans; D007473:Ion Channels / Q000378:metabolism*; D007834:Lasers; D018408:Patch-Clamp Techniques / Q000191:economics / Q000295:instrumentation* / Q000379:methods*; D051381:Rats; D017640:Silicates
D004129:Dimethylpolysiloxanes; D007473:Ion Channels; D017640:Silicates; C013830:baysilon