The patch clamp method measures membrane currents at very high resolution when a high-resistance 'gigaseal' is established between the glass microelectrode and the cell membrane (Pflugers Arch. 391 (1981) 85; Neuron 8 (1992) 605). Here we describe the first use of the silicone elastomer, poly(dimethylsiloxane) (PDMS), for patch clamp electrodes. PDMS is an attractive material for patch clamp re...

This course module introduces the analysis of data from intracellular patch-clamp recordings in the voltage clamp mode, focusing on the detection and statistical description of spontaneous as well as evoked post-synaptic currents (PSCs). After having completed the module, students should be able to detect PSCs by setting appropriate thresholds and analyze amplitude distribution, reliability and...

This protocol describes the loading of individual cells with fluorescent probes via patch pipettes. The patch-clamp methodology has been successfully used for single-cell dye labeling in cultured neurons, brain slices, and in vivo preparations. A broad range of dyes can be used with this loading technique. Markers for morphological reconstruction (e.g., Lucifer yellow); ion-sensitive indicator ...

Subthreshold potentials are thought to be mediated by time-independent, "passive" background currents. In this study, we show that the background current-voltage (I-V) relation of guinea pig ventricular myocytes is changed significantly by repetitive stimulation, in such a way that cell excitability becomes enhanced. Myocytes were used for whole-cell voltage-clamp experiments. A voltage-clamp r...

RESEARCH DESIGN AND METHODS—A total of 147 overweight (BMI$85th percentile for age and sex) youth, aged 8 to,20 years, undertook three tests: 1) a 3-h hyperinsulinemiceuglycemic clamp; 2) a 2-h hyperglycemic clamp; and 3) a 2-h OGTT. Participants were categorically assigned to five groups according to their OGTT 2-h plasma glucose level, ranging from,120 to $200 mg/dL. b-Cell function relative ...

The bacteriophage T4 DNA polymerase holoenzyme is composed of the core polymerase, gene product 43 (gp43), in association with the "sliding clamp" of the T4 system, gp45. Sliding clamps are the processivity factors of DNA replication systems. The T4 sliding clamp comes to encircle DNA via the "clamp loader" activity inherent in two other T4 proteins: 44 and 62. These proteins assemble into a pe...

Patch-clamp recording is undoubtedly the leading method for studying the behaviour of ion channels. In 1976, Neher and Sakmann (1) first used the cell-attached configuration of the patch-clamp technique to resolve current flow through single acetylcholine-activated cation channels in frog skeletal muscle. Subsequent improvements to the technique produced a range of methods for the high resoluti...