Sutter p 97 horizontal puller

Manufactured by Sutter Instruments

Sourced in United States

The Sutter P-97 horizontal puller is a device used to create micropipettes and other similar pulled glass instruments. It utilizes heat and precise control to draw and shape glass capillaries into the desired form.

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Lab products found in correlation

Axopatch 200b amplifier, by Molecular Devices (1 mentions) Pclamp8, by Molecular Devices (1 mentions) Axiovert 200, by Zeiss (1 mentions) Borosilicate glass, by Harvard Apparatus (1 mentions) Borosilicate glass, by World Precision Instruments (1 mentions) Pulse software, by HEKA Elektronik (1 mentions) Heka epc 10 amplifier, by HEKA Elektronik (1 mentions) Digidata 1440a, by Molecular Devices (1 mentions) Axopatch 700b amplifier, by Molecular Devices (1 mentions)

4 protocols using sutter p 97 horizontal puller

Whole-cell recording of Ca2+-activated Cl- currents

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The Ca²⁺-activated Cl⁻ channel currents were recorded with an Axopatch 200B Amplifier (Axon Instruments, Foster City, CA, USA) using the whole-cell recording technique at room temperature. Patch pipettes were made from borosilicate capillary tubes with a Sutter P-97 horizontal puller (Sutter Instrument, Novato, CA, USA). The resistance of the pipettes was 3–6 MΩ after filling with pipette solution. The currents were elicited with voltage steps from −100 mV to +100 mV in a +20 mV increment for 250 ms with an interval of 5 s from a holding potential of −50 mV. Currents were filtered at 2 kHz and sampled at 5 kHz using pCLAMP8.0 software (Axon Instruments). The extracellular solution contained (mM): NMDG-Cl 125, KCl 5, CaCl₂ 1.5, MgSO₄ 1, HEPES 10, glucose 10 and pH was adjusted to 7.4 with NMDG. The pipette solution contained (mM): CsCl 130, Mg·ATP 1, MgCl₂ 1.2, HEPES 10, EGTA 2, CaCl₂ 1.639 and pH was adjusted to 7.4 with CsOH. The intracellular Ca²⁺ concentration was 260 nM.

Liu C.Z., Li F.Y., Lv X.F., Ma M.M., Li X.Y., Lin C.X., Wang G.L, & Guan Y.Y. (2019). Endophilin A2 regulates calcium-activated chloride channel activity via selective autophagy-mediated TMEM16A degradation. Acta Pharmacologica Sinica, 41(2), 208-217.

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Patch-Clamp Recording of Action Potentials

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To record cellular action potentials we placed the cells in an experimental chamber (0.5 ml) mounted on the stage of an inverted microscope (Zeiss Axiovert 200, Carl Zeiss, Oberkochen, Germany). A room temperature bath solution (20–22°C) continuously perfused the chamber. Patch electrodes were pulled from borosilicate glass (Harvard Apparatus, Holliston, MA, USA) on a Sutter P-97 horizontal puller (Sutter Instruments, Novato, CA, USA). The fire-polished electrodes had a tip diameter of 2–3 μm and a resistance of ~2MΩ when filled with the patch electrode solution, which contained (in mM): 127 KCl, 10 Hepes, 10 NaCl, 0.1 cAMP, 5 MgATP. This solution was used for experiments for recording action potentials. We used a HEKA EPC-10 patch-clamp amplifier (HEKA, Bellmore, USA) which was controlled by PatchMaster v2x53 software (HEKA, Bellmore, USA).

Kaese S., Larbig R., Rohrbeck M., Frommeyer G., Dechering D., Olligs J., Schönhofer-Merl S., Wessely R., Klingel K., Seebohm G, & Eckardt L. (2017). Electrophysiological alterations in a murine model of chronic coxsackievirus B3 myocarditis. PLoS ONE, 12(6), e0180029.

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Electrophysiology of Glucose-Exposed GlyR-HEK Cells

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Current responses from GlyR-transfected HEK293 cells were measured at room temperature (21–23°C) at a holding potential of −50 mV using a HEKA EPC10 amplifier (HEKA Electronics, Lambrecht, Germany) controlled by Pulse software (HEKA Electronics). Recording pipettes were pulled from borosilicate glass (World Precision Instruments, Berlin, Germany) using a Sutter P-97 horizontal puller (Sutter, Novato, CA, USA). Solutions were applied using an Octaflow system (NPI electronics, Tamm, Germany). The external buffer consisted of 135 mM NaCl, 5.5 mM KCl, 2 mM CaCl₂, 1.0 mM MgCl₂, and 10 mM Hepes (pH adjusted to 7.4 with NaOH); the internal buffer was 140 mM CsCl, 1.0 mM CaCl₂, 2.0 mM MgCl₂, 5.0 mM EGTA, and 10 mM Hepes (pH adjusted to 7.2 with CsOH). Glucose (Sigma-Aldrich, Munich, Germany) was added to the growth medium on the day before experiments to give a pre-exposure to 50 mM of glucose for 16–20 h. Dose-response data were fitted to the Hill equation (see Supplementary Material) to determine EC₅₀ and IC₅₀. Significance of differences between EC₅₀ values were determined using one-way ANOVA with p ≤ 0.05 (*) and p ≤ 0.01 (**) taken as significant.

Hussein R.A., Ahmed M., Breitinger H.G, & Breitinger U. (2019). Modulation of Glycine Receptor-Mediated Pain Signaling in vitro and in vivo by Glucose. Frontiers in Molecular Neuroscience, 12, 280.

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Whole-cell Patch-clamp Recordings

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Membrane currents were measured using whole-cell patch-clamp procedures with Axopatch 700B amplifiers (Axon Instruments Foster City, CA). Patch electrodes were pulled from borosilicate glass (Hilgenberg) on a Sutter P-97 horizontal puller (Sutter Instruments, Novato, CA, USA). All signals were acquired at 0.33 kHz (Digidata 1440A, Axon Instruments) and analyzed with pCLAMP version 10.2 (Axon Instruments). Series resistance and capacitive transients were compensated using standard techniques. Membrane currents were low pass filtered at 10 kHz and digitized at 100 kHz. Recordings were made at room temperature using an internal solution containing (mmol/L) 140 KAsp, 10 EGTA, 4 MgATP, 1 MgCl2, 10 HEPES, (pH = 7.2 with KOH). External solution contained (mmol/L) 140 NaCl, 1 CaCl2, 1 MgCl2, 4 KCl. 10 HEPES, and 5 Glucose (pH = 7.4 with NaOH). In order to ensure reproducibility, after whole-cell conditions were established by rupturing the cell membrane, we allowed a dialysis period of 4 minutes before beginning measurements. During the dialysis period, we monitored current-voltage relationships to ensure stability and consistency of recordings.

Hu D., Li Y., Zhang J., Pfeiffer R., Gollob M.H., Healey J., Harrell D.T., Makita N., Abe H., Sun Y., Guo J., Zhang L., Yan G., Mah D., Walsh E.P., Leopold H.B., Giustetto C., Gaita F., Zienciuk-Krajka A., Mazzanti A., Priori S.G., Antzelevitch C, & Barajas-Martinez H. (2017). The Phenotypic Spectrum of a Mutation Hotspot Responsible for the Short QT Syndrome. JACC. Clinical electrophysiology, 3(7).

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