Nanion Technologies


Data and Applications

Accurate Voltage Clamp

SyncroPatch 384PE Nav17 IV

CHO cells expressing NaV1.7 were used on the SyncroPatch 384PE with a success rate of >90% for cells which have a seal resistance > 500 MΩ (see inset). A screenshot of the PatchControl 384 software showing current traces in response to a voltage step protocol and the corresponding current-voltage plot.  Cells were kindly provided by Anaxon.

Activation and Modulation of NMDA NR1/NR2A

NMDA 384well view

Here, activation and modulation of the NR1/NR2A subunit containing NMDA receptors are shown. Currents were evoked by application of 10 µM Glutamate in the presence of 10 µM Glycine.
Application of the neurosteroid Pregnenolone sulfate (PS) potentiates the current response induced by Glutamate. The potency of PS was analysed in a single-point screen. The EC50 of PS (39.6µM), comprising 74% of the cells is nicely corresponding to litereature values (Irwin, et al. Neurosci. Lett. 1992)
Cells were kindly provided by Bsys.

Activation of CFTR by internal fluoride or external forskolin

CFTR Data SyncroPatch 384

Image: Data acquisition during an experiment recording CFTR expressed in CHO cells. Half the plate (columns 1 – 12) had an internal solution without F- where CFTR was activated by external application of 20 µM forskolin, and half the plate (columns 13 – 24) had an internal solution containing 120 mM F-. On the right, raw current traces are shown to a voltage ramp protocol, in the absence (red trace) and presence of external forskolin (black and blue traces). The online analysis showing current amplitude at +/-100 mV versus time is also shown. The bottom panel shows activation of CFTR by internal perfusion from a solution containing 0 mM to 60 mM F-. Cells were kindly provided by Charles River.

Kv1.3 Pharmacology with High Success Rate

Kv13 384 raw onl norm Quini

Shown are screenshots of a pharmacology experiment performed with the 384PE. Recordings from 384 Kv1.3 expressing CHO cells were performed simultaneously. Original current traces and the peak current over time are displayed. Data are analysed with DataControl384 full analysis tool. With just a few mouse-clicks normalized concentration response curves can be generated. Here, normalized response and the IC50 of Quinidine is shown. Darkening shades of blue indicate increasing compound concentration.  Cells were kindly provided by Evotec.

Recording of Glutamate Receptor Currents

GluA2 repetitive CRC

Using a stacked solutions approach and a fast pipetting speed shortens the solution exchange rate and minimizes the ligand exposure time. This procedure allows for reproducible recordings of fast desensitizing ligand-gated receptors such as glutamate receptors. Here, repetitive activation of GluA2 receptors is shown. Receptors were activated with 100 µM Na-Glutamate for 3 times resulting in inward currents of similar peak amplitudes (A and B). The current onset time was approximately 10 ms (D). Panel C displays an example of a cumulative concentration response curve for Na-Glutamate (in mM: 0.1, 0.3 and 1). Cells were kindly provided by University of Sussex.

Reproducible Current Recordings

Glycine 384 raw OA reproducible

Glycine-mediated current traces and corresponding time plots from 384 simultaneously recorded HEK cells are shown. Multiple additions of 50 µM Glycine produce very robust current responses with similar peaks, providing best conditions for cumulative pharmacology on one cell.

Stable hERG recordings with accurate pharmacology

Syncro hERG 2

Current-voltage relationship of hERG (Kv11.1) expressed in HEK293 is shown along with pharmacology of 4 hERG-active compounds. The current-voltage relationships for all 384 wells (top) using perforated patch (Escin) and multi-hole chips (4 holes per well) are shown. In all 384 wells, a hERG-mediated current was observed with peak amplitude >700 pA at -20 mV. Using a pharmacology voltage protocol, experiments were stable lasting over 20 minutes. Concentration response curves for astemizole, pimozide, cisapride and terfenadine revealed IC50 values consistent with those found in the literature. Cells were kindly provided by Charles River Laboratories.