Microfabricated teflon membranes for low-noise recordings of ion channels in planar lipid bilayers

被引:194
作者
Mayer, M
Kriebel, JK
Tosteson, MT
Whitesides, GM
机构
[1] Harvard Univ, Dept Chem & Biol Chem, Cambridge, MA 02138 USA
[2] Harvard Univ, Sch Med, Dept Cell Biol, Cambridge, MA 02138 USA
关键词
D O I
10.1016/S0006-3495(03)74691-8
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
We present a straightforward, accessible method for the fabrication of micropores with diameters from 2 to 800 mum in films of amorphous Teflon (Teflon AF). Pores with diameters less than or equal to40 mum made it possible to record ion fluxes through ion channels in planar bilayers with excellent signal characteristics. These pores afforded: i), stable measurements at transmembrane voltages up to 460 mV; ii), recordings at low noise levels (0.4 pA rms at 4.3 kHz bandwidth); iii), recordings at high effective bandwidth (10.7 kHz); and iv), formation of multiple planar lipid bilayers in parallel. Microfabricated pores in films of Teflon AF made it possible to examine, experimentally and theoretically, the influence of the pore diameter on the current noise in planar bilayer recordings. Reducing the pore diameter below 40 mm mainly increased the stability of the planar bilayers, but had only a small effect on the level of the current noise. The low-noise properties of bilayer recordings on micropores in Teflon AF films were exploited to record the smallest conductance state of alamethicin (24 pS) at an unprecedentedly high bandwidth of 10.7 kHz.
引用
收藏
页码:2684 / 2695
页数:12
相关论文
共 47 条
[1]  
ALVAREZ O, 1986, ION CHANNEL RECONSTI, P115
[2]  
[Anonymous], FUOROPOLYMERS
[3]  
[Anonymous], 1976, Filtering in the Time and Frequency Domains
[4]  
Benndorf Klaus, 1995, P129
[5]   ALAMETHICIN PORE FORMATION - VOLTAGE-DEPENDENT FLIP-FLOP OF ALPHA-HELIX DIPOLES [J].
BOHEIM, G ;
HANKE, W ;
JUNG, G .
BIOPHYSICS OF STRUCTURE AND MECHANISM, 1983, 9 (03) :181-191
[6]   STATISTICAL-ANALYSIS OF ALAMETHICIN CHANNELS IN BLACK LIPID-MEMBRANES [J].
BOHEIM, G .
JOURNAL OF MEMBRANE BIOLOGY, 1974, 19 (03) :277-303
[7]   Simultaneous optical and electrical recording of single gramicidin channels [J].
Borisenko, V ;
Lougheed, T ;
Hesse, J ;
Füreder-Kitzmüller, E ;
Fertig, N ;
Behrends, JC ;
Woolley, GA ;
Schütz, GJ .
BIOPHYSICAL JOURNAL, 2003, 84 (01) :612-622
[8]  
Colquhoun David, 1995, P483
[9]   HTS approaches to voltage-gated ion channel drug discovery [J].
Denyer, J ;
Worley, J ;
Cox, B ;
Allenby, G ;
Banks, M .
DRUG DISCOVERY TODAY, 1998, 3 (07) :323-332
[10]   Whole cell patch clamp recording performed on a planar glass chip [J].
Fertig, N ;
Blick, RH ;
Behrends, JC .
BIOPHYSICAL JOURNAL, 2002, 82 (06) :3056-3062