Pc 10 pipette puller

Manufactured by Narishige

Sourced in Japan, United States

The Narishige PC-10 is a pipette puller designed to produce micropipettes from glass capillary tubes. It utilizes a heating element to melt the glass and gravity to draw the capillary into two separate pipettes.

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Lab products found in correlation

Multiclamp 700a amplifier, by Molecular Devices (2 mentions) Multiclamp 700b amplifier, by Molecular Devices (2 mentions) Lumplanfl n, by Olympus (1 mentions) Alexa fluor 594, by Thermo Fisher Scientific (1 mentions) Borosilicate glass with filament, by Sutter Instruments (1 mentions) P 885 piezoelectric stack actuator, by Physik Instrumente (1 mentions) Microforge mf 830, by Narishige (1 mentions) Bx51wi, by Olympus (1 mentions) Borosilicate glass capillary tubes, by Warner Instruments (1 mentions) Im 11 three dimensional mobile manipulator, by Narishige (1 mentions) Axopatch 200b patch clamp amplifier, by Molecular Devices (1 mentions) Ie190, by World Precision Instruments (1 mentions) Tw150f 4, by World Precision Instruments (1 mentions) Axon 200b amplifier, by Molecular Devices (1 mentions) Borosilicate glass capillary tubes, by Harvard Apparatus (1 mentions) Clampex 10, by Molecular Devices (1 mentions) Axio examiner a1 microscope, by Zeiss (1 mentions) Tc 344b, by Warner Instruments (1 mentions) Digidata 1550a digitizer, by Molecular Devices (1 mentions)

9 protocols using pc 10 pipette puller

Whole-cell voltage-clamp recordings of retinal ganglion cells

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Whole-cell voltage clamp recordings were made from whole-mount retinas continuously superfused in oxygenated ACSF (32–34°C) at a rate of 2–4 ml/min. Retinas were visualized under infrared illumination (870 nm). Voltage-clamp recordings from somas of ganglion cells (holding potential of −60/−65 mV) were obtained using glass microelectrodes of 4–5 MΩ (PC-10 pipette puller; Narishige, East Meadow, NY) filled with an internal solution containing (in mM): 110 CsMeSO₄, 2.8 NaCl, 4 EGTA, 5 TEA-Cl, 4 adenosine 5′-triphosphate (magnesium salt), 0.3 guanosine 5′-triphosphate (trisodium salt), 20 HEPES and 10 phosphocreatine (disodium salt), pH 7.2 and 290 mOsm. The liquid junction potential correction for this solution was −13 mV. Signals were acquired using pCLAMP 9 recording software and a Multiclamp 700 A amplifier (Molecular Devices, Sunnyvale, CA), sampled at 20 kHz and low-pass filtered at 2 kHz.
RGC dendritic stratification was visualized by including 20 μM Alexa Fluor 594 (Invitrogen, Grand Island, NY) in the intracellular solution. The dendritic morphology of dye-injected RGCs was reconstructed by two-photon imaging with the laser tuned to 780 nm. Images (RGCs and whole retina) were acquired at z intervals of 0.5 μm using a 60× objective (Olympus 60×, 1 NA, LUMPlanFLN). Images were later reconstructed from image stacks with ImageJ.

Rosa J.M., Bos R., Sack G.S., Fortuny C., Agarwal A., Bergles D.E., Flannery J.G, & Feller M.B. (2015). Neuron-glia signaling in developing retina mediated by neurotransmitter spillover. eLife, 4, e09590.

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Electrophysiological Recordings of Cochlear Hair Cells

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Cochlear hair cells were observed with an upright microscope (Olympus BX51WI). Borosilicate glass with filament (Sutter) was pulled with a PC-10 pipette puller (Narishige) and polished with MF-830 microforge (Narishige) to resistance of 3–5 MOhm. Hair bundles were deflected with a glass pipette mounted on a P-885 piezoelectric stack actuator (Physik Instrumente). Whole cell currents were sampled at 100 KHz with an EPC 10 USB patch-clamp amplifier operated by Patchmaster software (HEKA). Extracellular solution contains (in mM) 144 NaCl, 0.7 NaH₂PO₄, 5.8 KCl, 1.3 CaCl₂, 0.9 MgCl₂, 5.6 glucose, and 10 H-HEPES, pH 7.4. Intracellular solution contains (in mM) 140 KCl, 1 MgCl₂, 0.1 EGTA, 2 Mg-ATP, 0.3 Na-GTP, and 10 H-HEPES, pH 7.2. Hair cells were voltage-clamped at −70 mV.

Chen J., Zhang X., Li J., Song C., Jia Y, & Xiong W. (2016). Identification of a Novel ENU-Induced Mutation in Mouse Tbx1 Linked to Human DiGeorge Syndrome. Neural Plasticity, 2016, 5836143.

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Electrophysiological Profiling of Spiking HEK Cells

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Spiking HEK-293 cells were placed in the same bath solution as that used for QPI at room temperature (25 °C). Glass micropipettes were pulled from borosilicate glass capillary tubes (Warner Instruments) using a PC-10 pipette puller (Narishige) and were loaded with internal solution containing (in mM) 125 potassium gluconate, 8 NaCl, 0.6 MgCl₂, 0.1 CaCl₂, 1 EGTA, 10 HEPES, 4 Mg-ATP, and 0.4 Na-GTP (pH 7.3, adjusted with NaOH; 295 mOsm, adjusted with sucrose). The resistance of the pipettes filled with internal solution varied between 2 and 3 MΩ. After setting up the whole cell configuration, the membrane potential during the spontaneous action potentials of spiking HEK cells was monitored with a Multiclamp 700B amplifier (Molecular Devices) under the current clamp mode.

Ling T., Boyle K.C., Goetz G., Zhou P., Quan Y., Alfonso F.S., Huang T.W, & Palanker D. (2018). Full-field interferometric imaging of propagating action potentials. Light, Science & Applications, 7, 107.

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Extraction and Analysis of Lipid Droplets

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A PC-10 pipette puller (Narishige Group, Tokyo, Japan) was used for preparing the borosilicate glass nanotips (length 55 mm, tip i.d. 2–6 μm, capillary i.d. 0.6 mm) according to the manufacturer’s instructions. The 1st step heating temperature was set at 60.5 °C, and the 2nd step heating temperature was set at 42−50 °C, depending on the tip diameter. The diameter of each prepared nanotip was confirmed under the microscope.
The LD sampling was conducted using an IM-11 three-dimensional mobile manipulator (Narishige Group), coupled with an IX71 inverted microscope in a bright field, as in our previous studies [24 (link),27 (link)]. The culture medium of the cells was washed and then replaced with 0.6 mL 160 mM ammonium formate solution, and the LDs were aspirated into nanotips using the microinjector. The obtained single LD was immediately extracted with 10 µL of methanol/isopropanol (1:9 v/v, with 0.1% trifluoroacetate).

Chen Z., Shrestha R., Yang X., Wu X., Jia J., Chiba H, & Hui S.P. (2022). Oxidative Stress and Lipid Dysregulation in Lipid Droplets: A Connection to Chronic Kidney Disease Revealed in Human Kidney Cells. Antioxidants, 11(7), 1387.

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Voltage-Dependent FM 4-64 SHG/F Ratio

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The voltage dependence of the FM 4-64 generated SHG/F ratio signal was obtained with a combination of whole cell patch-clamp and SHG microscopy. Whole-cell recordings were obtained using an Axopatch 200B patch-clamp amplifier (Molecular Devices) and uncoated borosilicate glass pipettes. Pipettes were pulled on a Narishige PC-10 pipette puller with resistances of 6-8 MOhm. Currents, which were not analyzed for this study, were low-pass filtered at 2.8 kHz with an 8-pole Bessel filter and digitized at 10 kHz. Recordings were made from visually identified AMs or J774 cells at +37°C, at V_h = −40 mV. The extracellular bath solution contained (in mM): 125 NaCl, 2.5 KCl, 10 HEPES, 1.5 MgCl₂, 2.5 CaCl₂, 6 D-glucose, 28 Sucrose to adjust osmolarity to 310 mOsm, pH 7.3 was adjusted with NaOH (Na extracellular solution). The patch pipette contained (in mM): 135 KCl, 4 NaCl, 1 MgCl₂, 10 HEPES, 2 Mg-ATP pH 7.2 adjusted with KOH, osmolarity adjusted to 280 mOsm with Sucrose.

Riazanski V., Mauleon G., Zimnicka A.M., Chen S, & Nelson D.J. (2021). Phagosomal chloride dynamics in the alveolar macrophage. iScience, 25(1), 103636.

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Fabrication of Potassium-Sensitive Microelectrodes

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Borosilicate glass capillaries (World Precision Instruments TW150F-4) were washed for 6 h with 1 M HCl and rinsed with 70% Ethanol, followed by incubation @ 120 °C overnight. Capillaries were then stored in an airtight container with anhydrous calcium sulfate desiccant until ready for pulling. Tips were created with a Narishige PC-10 pipette puller set to 84.8 and 61.2. Tips were then placed in a glass coplin jar and silanized by applying 50 µL of dicholorodimethylsilane (Tokyo Chemical Industry B2150) below the tips and sealed. The coplin jars were then placed in an oven @ 200 °C for 30 min. Following silanization, tips were visualized under a light microscope and tips manually broken to a size of ~5−10 µM. Tips were then loaded with 2 µL of potassium sensitive ionophore (World Precision Instruments IE190) and backfilled with KCl (100 mM). Microelectrodes were stored with tips down in a solution of KCl (100 mM) for up to ~1 h prior to use.

Cleary C.M., Browning J.L., Armbruster M., Sobrinho C.R., Strain M.L., Jahanbani S., Soto-Perez J., Hawkins V.E., Dulla C.G., Olsen M.L, & Mulkey D.K. (2024). Kir4.1 channels contribute to astrocyte CO2/H+-sensitivity and the drive to breathe. Communications Biology, 7, 373.

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Voltage-clamp Recordings from Retinal GCs

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Whole-cell voltage-clamp recordings were made from whole-mount retinas while simultaneously imaging GCaMP6f fluorescence. Under infrared illumination, RGC somas were targeted for voltage-clamp recordings using glass microelectrodes with resistance of 3–5 MΩ (PC-10 pipette puller; Narishige) filled with an internal solution containing (in mM) 110 CsMeSO₄, 2.8 NaCl, 20 HEPES, 4 EGTA, 5 TEA-Cl, 4 Mg-ATP, 0.3 Na₃GTP, 10 Na₂Phosphocreatine, and QX-Cl (pH 7.2 and 290 mOsm). The liquid junction potential correction for this solution was –10 mV. Signals were acquired using pCLAMP10 recording software and a MultiClamp 700A amplifier (Molecular Devices), sampled at 20 kHz and low-pass filtered at 2 kHz.

Tworig J.M., Coate C.J, & Feller M.B. (2021). Excitatory neurotransmission activates compartmentalized calcium transients in Müller glia without affecting lateral process motility. eLife, 10, e73202.

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TMEM16 Channel Currents Measurement

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TMEM16s currents were measured with the whole-cell configuration of the patch-clamp technique using an Axon 200B amplifier (Molecular Devices, USA) controlled with GE-pulse software (http://users.ge.ibf.cnr.it/pusch/programs-mik.htm). Currents were low-pass filtered at 2 kHz and sampled at 10 kHz. Pipettes were prepared from borosilicate glass capillary tubes (Harvard Apparatus, USA) using a PC-10 pipette puller (Narishige, Japan). Pipette tip diameter yielded a resistance of ~2–3 MΩ in the working solutions. The bath was grounded through a 3 M KCl agar bridge connected to a Ag-AgCl reference electrode. Experiments were conducted at 20–22 °C. The cell capacitance was assessed by measuring the area beneath a capacitive transient elicited by a 10 mV step or via the cell capacity compensation circuit of the amplifier. Current density was obtained by dividing the current amplitude for the cell capacitance.
The extracellular solution contained: 150 mM NaCl, 1 mM CaCl₂, 1 mM MgCl₂, 10 mM glucose, 10 mM d-mannitol and 10 mM HEPES; pH was adjusted to 7.4 with NaOH. The intracellular solution contained: 130 mM CsCl, 10 mM EGTA, 1 mM MgCl₂, 10 mM HEPES and 8 mM CaCl₂ to obtain [Ca²⁺]_i of ~300 nM; pH was adjusted to 7.3 with NaOH. The intracellular solution containing ~78 μM [Ca²⁺]_i was obtained by replacing EGTA with equimolar H-EDTA and by adding a total of 9 mM CaCl₂^{51 (link)}.

Bushell S.R., Pike A.C., Falzone M.E., Rorsman N.J., Ta C.M., Corey R.A., Newport T.D., Christianson J.C., Scofano L.F., Shintre C.A., Tessitore A., Chu A., Wang Q., Shrestha L., Mukhopadhyay S.M., Love J.D., Burgess-Brown N.A., Sitsapesan R., Stansfeld P.J., Huiskonen J.T., Tammaro P., Accardi A, & Carpenter E.P. (2019). The structural basis of lipid scrambling and inactivation in the endoplasmic reticulum scramblase TMEM16K. Nature Communications, 10, 3956.

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Whole-Cell Patch-Clamp Recording from Hippocampal CA1 Neurons

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Slices were continuously perfused with extracellular recording solution warmed to 30–34°C with an external heating apparatus (Warner Instruments, TC-344B). The CA1 subfield of the hippocampus was visualized using a 4× magnification objective on a Zeiss Axio Examiner A1 microscope (Zeiss). Magnification was switched to a 63× fluid-immersion objective to identify putative pyramidal neurons in the CA1 pyramidal layer. Pipettes for whole-cell patch-clamp physiology were pulled (Narishige PC-10 pipette puller; Narishige) from borosilicate glass pipettes (WPI #1B1505-4, World Precision Instruments) to a resistance of 2–6 MΩ. Pipettes were filled with an intracellular solution containing 135 mm KMeSO₄, 10 mm HEPES, 2 mm MgATP, 0.1 mm NaGTP, 8 mm NaCl, 0.1 mm BAPTAK₄, and 0.2% biocytin (pH 7.25). Membrane potentials were recorded using a MultiClamp 700B amplifier (Molecular Devices), processed using a Digidata 1550A digitizer, and analyzed using Clampex 10.4 software (Molecular Devices) on a Microsoft Windows-based computer. Membrane potentials were observed in response to stepwise, 25-pA current increases from −100 to 400 pA.

Marks W.D., Paris J.J., Barbour A.J., Moon J., Carpenter V.J., McLane V.D., Lark A.R., Nass S.R., Zhang J., Yarotskyy V., McQuiston A.R., Knapp P.E, & Hauser K.F. (2021). HIV-1 Tat and Morphine Differentially Disrupt Pyramidal Cell Structure and Function and Spatial Learning in Hippocampal Area CA1: Continuous versus Interrupted Morphine Exposure. eNeuro, 8(3), ENEURO.0547-20.2021.

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