Log In Sign up
The largest database of trusted experimental protocols

Pc 10 vertical puller

Manufactured by Narishige
Visit Supplier
Sourced in United States

The Narishige PC-10 vertical puller is a laboratory instrument designed for pulling glass micropipettes. It utilizes a vertical heating element to melt and pull the glass, creating pipettes with precise and consistent tip diameters.

Automatically generated - may contain errors

Lab products found in correlation

Pclamp 10, by Molecular Devices (3 mentions) Axopatch 1b, by Molecular Devices (2 mentions) Digidata 1440a, by Molecular Devices (1 mentions) Borosilicate glass capillaries, by Harvard Apparatus (1 mentions) Spermine tetrahydrochloride, by Merck Group (1 mentions) Digidata 1440a interface, by Molecular Devices (1 mentions) Axopatch 200b amplifier, by Molecular Devices (1 mentions) Borosilicate glass, by Harvard Apparatus (1 mentions) Tc 344b, by Warner Instruments (1 mentions) Vt1200s vibratome, by Leica (1 mentions) Multiclamp 700b, by Molecular Devices (1 mentions) D apv, by Abcam (1 mentions) Picrotoxin, by Abcam (1 mentions) Kynurenic acid, by Abcam (1 mentions) Multiclamp 700b amplifier, by Molecular Devices (1 mentions) Digidata 1440, by Molecular Devices (1 mentions) Alexa 488 dye, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

PC-10 (102 citations) PC-10 puller (37 citations) Vertical puller (25 citations) PC-10 vertical Micropipette Puller (2 citations)

7 protocols using pc 10 vertical puller

1

Cardiomyocyte Electrophysiology Recordings

Check if the same lab product or an alternative is used in the 5 most similar protocols
Raptured whole-cell patch clamp recordings from visually identified cardiomyocytes were performed using SliceScope (Scientifica, Uckfield, UK) equipped with a CCD camera. Recordings were done with PC505B amplifier (Warner Instruments, CT, USA) in voltage and current clamp modes. Signals were filtered at 2 kHz with the amplifier, then acquired and digitized at 10 kHz sampling frequency with Digidata 1440A (Molecular Devices, CA, USA) and software package for data acquisition and analysis pClamp 10.2 (Molecular Devices, CA, USA). Recording electrodes of 4–5 MOhm resistance were pulled from borosilicate glass capillaries (Harvard Apparatus, MA, USA) using a PC10 vertical puller (Narishige, NY, USA). Extracellular Hank’s solution contained (in mM): 139 NaCl, 4.2 NaHCO3, 0.4 NaH2PO4, 2.1 KCl, 0.44 KH2PO4, 1.25 CaCl2, 0.8 MgSO4, 4 HEPES, 8 D-glucose, pH 7.4, and osmolality 305 ± 2 mOsm. The composition of the intracellular solution was as follows (in mM): 120 K-gluconate, 3 KCl, 2 Na2ATP, 0.3 Na2GTP, 0.3 MgATP, 10 Na2-phosphocreatine, 1 MgCl2, 0.25 EGTA, 4 HEPES, pH 7.2 and osmolality of 280 ± 5 mOsm. All experiments were conducted at 30 °C. Access resistance was monitored throughout the recording and was typically < 35 MOhm.
Averin A.S., Konakov M.V., Pimenov O.Y., Galimova M.H., Berezhnov A.V., Nenov M.N, & Dynnik V.V. (2022). Regulation of Papillary Muscle Contractility by NAD and Ammonia Interplay: Contribution of Ion Channels and Exchangers. Membranes, 12(12), 1239.
+ Open protocol
+ Expand
2

Whole-Cell Patch-Clamp Recordings of Macroscopic Currents

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were visualized with an inverted microscope (IX50; Olympus). Electrodes were fabricated from borosilicate glass (1.5 mm o.d., 0.86 mm i.d., Harvard Apparatus) pulled with a PC-10 vertical puller (Narishige). Electrode resistance varied between configurations (see below). Macroscopic currents were recorded at room temperature (22-25°C) in the whole-cell configuration (wc) or from outside-out patches (o) excised from GFP-positive cells. Currents were recorded with Axopatch 200B amplifier, filtered at 2 kHz (wc) or 10 kHz (o) and digitized at 5 kHz (wc) or 50 kHz (o) using Digidata 1440A interface with pClamp 10 software (Molecular Devices Corporation). For all configurations the “extracellular” solution contained (in mM): 145 NaCl, 2.5 KCl, 1 CaCl2, 1 MgCl2, 10 glucose and 10 HEPES (pH to 7.42 with NaOH). For fast agonist application, 10 mM glutamate was added to the “extracellular” solution. The “intracellular” solution contained (in mM): 145 CsCl, 2.5 NaCl, 1 Cs-EGTA, 4 MgATP, and 10 HEPES (pH to 7.2 with CsOH). Spermine tetrahydrochloride (Sigma Aldrich) was added to intracellular solution at 100 μM in all cases.
Gratacòs-Batlle E., Yefimenko N., Cascos-García H, & Soto D. (2015). AMPAR interacting protein CPT1C enhances surface expression of GluA1-containing receptors. Frontiers in Cellular Neuroscience, 8, 469.
+ Open protocol
+ Expand
3

Patch-Clamp Recordings of Striatal MSNs

Check if the same lab product or an alternative is used in the 5 most similar protocols
Fresh coronal NAcore slices (250 μm; VT1200S Leica vibratome) were collected into a vial containing aCSF (in mM: 126 NaCl, 1.4 NaH2PO4, 25 NaHCO3, 11 glucose, 1.2 MgCl2, 2.4 CaCl2, 2.5 KCl, 2.0 sodium pyruvate, 0.4 ascorbic acid, bubbled with 95% O2 and 5% CO2). For cutting and storage, a mixture of 5 mM kynurenic acid (abcam) and 50 μM D-APV (abcam) was added to the aCSF. Slices were kept at 22°C to 24°C until they were used for recordings and were constantly perfused with oxygenated aCSF heated to 32°C (TC-344B, Warner Instruments). GABAA synaptic transmission was blocked with 100 μM picrotoxin (abcam). Neurons were visualized with a Zeiss Axioscope 2 FS plus microscope with a 40× objective and voltage clamp recordings (Multiclamp 700B, Molecular Devices) performed from visualized MSNs in the medial NAcore near the anterior commissure. Glass microelectrodes (1.5–2.5 ΩM) were prepared using a PC-10 vertical puller (Narishige) and filled with internal solution as follows (in mM: 124 cesium methanesulfonate, 10 HEPES potassium, 1 EGTA, 1 MgCl2, 10 NaCl, 2.0 MgATP, and 0.3 NaGTP, 1 QX-314, pH 7.2–7.3, 290 mOsm). To evoke postsynaptic currents, a bipolar stimulating electrode was placed 300 μm dorso-medial of the recorded cell. Data was acquired at 10 kHz and filtered at 2 kHz using AxographX software (Axograph Scientific).
Neuhofer D., Spencer S.M., Chioma V.C., Beloate L.N., Schwartz D, & Kalivas P.W. (2019). The loss of NMDAR-dependent LTD following cannabinoid self-administration is restored by positive allosteric modulation of CB1 receptors. Addiction biology, 25(6), e12843.
+ Open protocol
+ Expand
4

Single-Channel Analysis of ENaC Activity

Check if the same lab product or an alternative is used in the 5 most similar protocols
ENaC activity was recorded using standard single channel patch clamp analysis in the cell-attatched configuration. Micropipettes were pulled with a two-stage Narishige PC-10 vertical puller (Narishige International, Amityville, NY) from filamented borosilicate glass capillaries purchased from World Precision Instruments (Sarasota, FL). Micropipette resistances were between 5–9 MΩ when filled and immersed in patch solution composed of (in mM): 140 NaCl, 5 KCl,1 CaCl2, 1 MgCl2, and 10 HEPES, adjusted to pH 7.40 with NaOH. Channel currents were sampled at 1 kHz with an Axopatch 1B patch amplifier (Molecular Devices) and filtered at 200 Hz with a low-pass Bessel filter. Continuous single channel activity was recorded and then analyzed using Pclamp 10 software for channel open probability (Po) and number (N) of active channels in SAEC apical membrane. Chord conductances (γ), were calculated from hyperpolarized and depolarized potentials.
Grant G.J., Coca C., Zhao X.M, & Helms M.N. (2020). Oxidized Glutathione Increases Delta-Subunit Expressing Epithelial Sodium Channel Activity in Xenopus laevis Oocytes. eMedical research, 2, 100008.
+ Open protocol
+ Expand
5

Single-Channel Analysis of ENaC Activity

Check if the same lab product or an alternative is used in the 5 most similar protocols
ENaC activity was recorded using standard single channel patch clamp analysis in the cell-attatched configuration. Micropipettes were pulled with a two-stage Narishige PC-10 vertical puller (Narishige International, Amityville, NY) from filamented borosilicate glass capillaries purchased from World Precision Instruments (Sarasota, FL). Micropipette resistances were between 5–9 MΩ when filled and immersed in patch solution composed of (in mM): 140 NaCl, 5 KCl,1 CaCl2, 1 MgCl2, and 10 HEPES, adjusted to pH 7.40 with NaOH. Channel currents were sampled at 1 kHz with an Axopatch 1B patch amplifier (Molecular Devices) and filtered at 200 Hz with a low-pass Bessel filter. Continuous single channel activity was recorded and then analyzed using Pclamp 10 software for channel open probability (Po) and number (N) of active channels in SAEC apical membrane. Chord conductances (γ), were calculated from hyperpolarized and depolarized potentials.
Grant G.J., Coca C., Zhao X.M, & Helms M.N. (2020). Oxidized Glutathione Increases Delta-Subunit Expressing Epithelial Sodium Channel Activity in Xenopus laevis Oocytes. eMedical research, 2, 100008.
+ Open protocol
+ Expand
6

Electrophysiological Characterization of Cholinergic Neurons in the Basal Forebrain

Check if the same lab product or an alternative is used in the 5 most similar protocols
Positive and negative ChAT-ChR2 neurons in the BF (HDB/MCPO) were visually identified by EYFP expression and differential interference contrast (DIC) on a modified Olympus upright microscope (Scientifca, East Sussex, United Kingdom). Whole cell recording was performed with a Multiclamp 700B amplifier (Molecular Devices Corp.), using recording pipettes with resistance of 3–5 MΩ pulled on a PC10 vertical puller (Narishige International) and filled with intracellular solution containing the following (in mM): 135 potassium gluconate, 20 KCl, 10 HEPES, 0.1 ethylene glycol tetraacetic acid (EGTA), 2 MgATP, and 0.3 NaGTP. Recordings were low-pass filtered at 4 kHz (Bessel filter) and digitized at 10 kHz (Digidata 1440) using pClamp 10.3 software (Molecular Devices Corp.). Series resistances were monitored throughout each voltage-clamp recording with 50 ms and −10 mV steps, and if it changed by >20%, the data were discarded. Evoked synaptic responses were recorded from ChAT+ and ChAT– neurons, and these responses were triggered by light stimulation directly onto the BF area. Light stimulation was evoked by a single mercury-free LED illumination system (CoolED pE-100 series) at 470 nm for 5 ms between 1 and 5% of the maximal intensity of the system. Latencies of evoked responses were analyzed using prewritten code routines in Axograph-X.
Nunez-Parra A., Cea-Del Rio C.A., Huntsman M.M, & Restrepo D. (2020). The Basal Forebrain Modulates Neuronal Response in an Active Olfactory Discrimination Task. Frontiers in Cellular Neuroscience, 14, 141.
+ Open protocol
+ Expand
7

Patch Clamp Recordings in Anesthetized Mice

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cell-attached and whole-cell patch clamp recordings were performed on mice under 1 – 1.5% isoflurane anesthesia on a 37ºC heating pad (DC Temperature Control System, FHC). Patch pipettes with resistance values between 5 – 7.5 MΩ were prepared by pulling filamented borosilicate glass capillaries (Warner or WPI) using a micropipette puller (Flaming-Brown P97 model, Sutter Instruments or PC-10 vertical puller, Narishige). These pipettes were filled with an internal solution containing (in mM): 135 K-gluconate, 4 KCl, 10 HEPES, 10 Na2-phosphocreatine, 4 MgATP, 0.3 Na3GTP (pH adjusted to 7.3 – 7.4 with KOH; osmolarity 280 – 290 mOsm), and 50 μM Alexa 488 dye (ThermoFisher; for pipette visualization under the two-photon microscope) or 125 K-Methanesulfonate, 7 KCl, 10 HEPES, 2 MgATP, 2 Na2ATP, 0.5 Na2GTP, 0.05 EGTA (pH adjusted to 7.3 with KOH; osmolarity 280–290 mOsm), and 50 μM Alexa 488 dye. Fully manual patch clamp experiments (Figures S4A–S4C) were performed following previously reported protocols (Häusser and Margrie, 2014 (link); Komai et al., 2006 (link)).
Suk H.J., van Welie I., Kodandaramaiah S.B., Allen B., Forest C.R, & Boyden E.S. (2017). Closed-loop real-time imaging enables fully automated cell-targeted patch clamp neural recording in vivo. Neuron, 95(5), 1037-1047.e11.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!