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P 87 puller

Manufactured by Sutter Instruments
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Sourced in United States

The P-87 puller is a precision instrument designed for the purpose of pulling glass micropipettes. It utilizes heat and precise control of mechanical parameters to generate micropipettes of consistent and customizable size and geometry.

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Lab products found in correlation

Axopatch 200b amplifier, by Molecular Devices (2 mentions) Digidata 1200, by Molecular Devices (1 mentions) Axopatch 200 amplifier, by Molecular Devices (1 mentions) Clampfit 10, by Molecular Devices (1 mentions) Prism 7 for windows, by GraphPad (1 mentions) Axon digidata 1322a, by Molecular Devices (1 mentions) 35 mm glass bottom petri dish, by MatTek (1 mentions) Digidata 1322a, by Molecular Devices (1 mentions) Pclamp 9, by Molecular Devices (1 mentions) Borosilicate glass, by Sutter Instruments (1 mentions)

5 protocols using p 87 puller

1

Whole-cell Voltage and Current Clamp Recordings

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Whole cell recordings were made using an Axopatch 200B amplifier and Digidata 1200 (Molecular Devices), as reported previously59 (link). Voltage clamp recordings were performed using a ramp protocol from −120 mV to +40 mV with cells held at −70 mV during the inter-pulse interval. The bath solution was Tyrode’s solution contained 137 mM NaCl, 5.4 mM KCl, 1 mM MgCl2, 2 mM CaCl2, 0.33 mM NaH2PO4, 5 mM HEPES and 1 mM Glucose with pH adjusted to 7.4 mM with NaOH. The pipette solution contained 120 mM KCl, 10 mM HEPES and 1 mM K-EGTA, with pH adjusted to 7.4. For current clamp recordings current was held at 0pA. The bath solution used for current clamp recordings contained 10 mM glucose for cells from islets chronically incubated in 30 mM glucose and was unaltered for cells from islets chronically incubated at 3 mM glucose. Glass electrodes were pulled from Kimble-Chase 2502 plain capillary tubes using a P-87 puller (Sutter instruments). For voltage clamp recordings electrodes with 1–3 MΩ tips were used and for current clamp recordings 5–7 MΩ tips were used.
Shyr Z.A., Wang Z., York N.W., Nichols C.G, & Remedi M.S. (2019). The role of membrane excitability in pancreatic β-cell glucotoxicity. Scientific Reports, 9, 6952.
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2

Local Field Potential Recording in Submerged Slices

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Local Field Potential (LFP) recordings were done in a submerged chamber, and slices were placed on a mesh that allowed perfusion on both sides at a high flow rate (10–30 ml/min) (Hajos and Mody, 2009 (link); Maier et al., 2009 (link)). All recordings were done with low resistance glass microelectrodes (∼150 kΩ tip resistance). The electrodes were pulled with a Sutter P87 puller with six controlled pulls and filled with 0.5 M NaCl in 1% agar, which prevents leakage of the electrode solution that could potentially alter the tissue surrounding the electrode tip. The recording electrode was placed in CA1 stratum pyramidale, where SWs have large amplitudes (Maier et al., 2009 (link)) in healthy slices.
Li P., Geng X., Jiang H., Caccavano A., Vicini S, & Wu J.Y. (2019). Measuring Sharp Waves and Oscillatory Population Activity With the Genetically Encoded Calcium Indicator GCaMP6f. Frontiers in Cellular Neuroscience, 13, 274.
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3

Hippocampal Slice SWR Signal Recording

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Low resistance glass microelectrodes were used for LFP recordings of the SW/R signals. The electrodes were pulled with a Sutter P87 puller with 6 controlled pulls, resulting in an approximate 150K tip resistance. Electrodes were filled with 1M NaCl in 1% agar, which prevents leakage of the electrode solution that could potentially alter conditions at the recording site. The recordings were done in a submerged chamber, and slices were perfused on both sides at a high flow rate (20 ml /min). In our recording arrangement SWR signals have an adequate signal-to-noise ratio. Hippocampal slices from dorsal and ventral locations demonstrated SWR activity. Comparisons between control and treated slices were performed with matched left and right slices of specific dorsal to ventral level sections (980–2450 microns from the dorsal most section). Thus, recordings were performed using control and treated pairs prepared at the same time and with the same cutting or recording ACSF.
Sun Z., Bozzelli P.L., Caccavano A., Allen M., Balmuth J., Vicini S., Wu J.Y, & Conant K. (2017). Disruption of perineuronal nets increases the frequency of sharp wave ripple events. Hippocampus, 28(1), 42-52.
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4

Patch-clamp recording of electrophysiology

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Patch pipettes (2.5–3 MΩ resistance) were fabricated from borosilicate glass capillaries (Friedrich & Dimmonck, Millville, New Jersey, USA) using a P-87 puller (Sutter Instruments, Novato, California, USA). Data were acquired and recorded with Clampex 10.3 software using an Axopatch 200 amplifier and Axon Digidata® 1322A (Molecular Devices, Sunnyvale, California, USA). Data were digitized at 20 kHz during all voltage protocols and a bandwidth of 2 kHz was set on the amplifier. For whole-cell recordings, cells were continuously perfused with extracellular solution at 33 ± 1 °C. Access resistance (Ra) was always below 5 MΩ and series resistance was typically compensated by ∼40%. Voltage protocols are described in the “Results” section. Patch-Clamp recordings were analysed using Clampfit 10.3 (Molecular devices, Sunnyvale, USA). Statistics and graphs were prepared using Excel Professional Plus 2013 (Microsoft Corporation, Redmond, Washington, USA) and Prism 7 for Windows (GraphPad Software, La Jolla, California, USA).
Lainez S., Doray A., Hancox J.C, & Cannell M.B. (2018). Regulation of Kv4.3 and hERG potassium channels by KChIP2 isoforms and DPP6 and response to the dual K+ channel activator NS3623. Biochemical Pharmacology, 150, 120-130.
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5

Single-Channel Recordings of Proteoliposomes

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We placed a 20 µL drop of proteoliposome suspension in the center of a 35 mm glass-bottom Petri dish (MatTek, Ashland, MA) that we had pretreated with 5% milk (w/v dry nonfat milk powder in phosphate buffered saline) for 30 min. Then we covered the dish with 2 mL of bath solution while taking care to keep the giant proteoliposomes near the center of the dish. Most of the giant proteopliposomes settled to the bottom of the dish within a few minutes. The bath solution consisted of 130 mM KCl, 1 mM MgCl2, and 10 mM HEPES with a pH titrated to 7.2 using KOH. The pipette solution was identical to the bath solution supplemented with 5 mM ATP. We fabricated the patch electrodes with resistances of 5.0–10.0 MΩ from borosilicate glass (Sutter Instruments, Novato, CA) using a P-87 puller (Sutter Instruments, Novato, CA). After formation of a Giga seal, we pulled the patch electrode away from the giant proteoliposome and quickly went through the water–air interface in order to ensure the inside-out configuration. We recorded the single channel current at room temperature using an Axopatch 200B amplifier (Molecular Devices, Sunnyvale, CA) and digitized the recordings using a Digidata 1322A (Molecular Devices, Sunnyvale, CA). The cut-off filter frequency was 5 kHz and the sampling rate was 25 kHz. Data acquisition was done using pClamp9 software (Molecular Devices, Sunnyvale, CA).
Horger K.S., Liu H., Rao D.K., Shukla S., Sept D., Ambudkar S.V, & Mayer M. (2014). Hydrogel-assisted functional reconstitution of human P-glycoprotein (ABCB1) in giant liposomes. Biochimica et biophysica acta, 1848(2), 643-653.
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