Borosilicate pipettes

Manufactured by Sutter Instruments

Sourced in United States

Borosilicate pipettes are precision laboratory instruments designed for accurate liquid handling. They are manufactured from borosilicate glass, a material known for its durability and resistance to chemical corrosion. These pipettes provide reliable and consistent volume delivery for a variety of applications in scientific research and clinical settings.

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Lab products found in correlation

Pclamp 10, by Molecular Devices (6 mentions) Axopatch 200b amplifier, by Molecular Devices (4 mentions) Lambda dg4 fluorescent excitation source, by Sutter Instruments (2 mentions) P 97 flaming brown puller, by Sutter Instruments (2 mentions) Axio observer, by Zeiss (2 mentions) Orca flash 4.0 camera, by Hamamatsu Photonics (2 mentions) Axopatch 700b amplifier, by Molecular Devices (1 mentions) X60 water immersion objective, by Nikon (1 mentions) Multiclamp 700b amplifier, by Molecular Devices (1 mentions)

9 protocols using borosilicate pipettes

Whole-cell electrophysiology recording protocol

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Recordings were made in whole-cell voltage (holding potential (V_h) of –60mV) or current clamp configurations at 22–24°C. Data were acquired using an Axopatch 200B amplifier and analyzed with pCLAMP10.2 software (Molecular Devices, Sunnyvale, CA). Recording data were filtered at 0.5–5 kHz and sampled at 2–20 kHz depending on current kinetics. Borosilicate pipettes (Sutter, Novato, CA) were polished to resistances of 2–3 MΩ. If required, access resistance (R_s) was compensated (40–80%) to the value of 6 MΩ. Data were rejected when R_s changed >20% during recording, leak currents were > 100pA, or input resistance was < 100 MΩ. Currents were considered positive when their amplitudes were 5-fold bigger than displayed noise (in root mean square). Standard external solution (SES) contained (in mM): 140 NaCl, 5 KCl, 2 CaCl₂, 1 MgCl₂, 10 D-glucose and 10 HEPES, pH 7.4. The standard pipette solution (SIS) contained (in mM): 140 KCl, 1 MgCl₂, 1 CaCl₂, 10 EGTA, 10 D-glucose, 10 HEPES, pH 7.3, 2.5 ATP and 0.2 GTP. Drugs were applied by a fast, pressure-driven and computer controlled 4-channel system (ValveLink8; AutoMate Scientific, San Francisco, CA) with quartz application pipettes.

Patil M.J., Hovhannisyan A.H, & Akopian A.N. (2018). Characteristics of sensory neuronal groups in CGRP-cre-ER reporter mice: Comparison to Nav1.8-cre, TRPV1-cre and TRPV1-GFP mouse lines. PLoS ONE, 13(6), e0198601.

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Calcium Imaging of Erythrocytes

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Erythrocyte calcium imaging experiments were performed as previously described15 (link). Briefly, blood was drawn and stored on ice in EGTA vacutainers. On the experimental day, we diluted whole blood 1:1,000 into NECM supplemented with 0.1% BSA and loaded with 5 μM Fluo-4 at 4 °C for at least an hour. Cells were then washed with buffer to remove excess dye and allowed to settle in a custom-build low-volume perfusion chamber, where a gravity-driven whole-chamber perfusion system was used to deliver Yoda1 and the calcium ionophore A23187 after erythrocytes loosely adhered to the uncoated glass-bottom chamber. For mechanical stimulation, we used long, tapered-tip (∼1.5–2 μm diameter) borosilicate pipettes (Sutter instruments, Novato, CA) forged with a Flaming/Brown P-97 puller (Sutter Instruments) and applied negative pressure to the cells using a High-speed Pressure Clamp device (ALA scientific, Farmingdale, NY). Imaging was performed using an Axio Observer (Zeiss) microscope fitted with a lambda DG4 fluorescent excitation source (Sutter Instruments). Images were recorded with an Orca Flash 4.0 camera (Hamamatsu Photonics) using MicroManager imaging software33 . Image analysis was performed with FIJI34 (link). HEK-P1KO cells were loaded with 2 μM Fura2-AM for 30 min at room temperature in NECM and imaged using the equipment described above.

Lukacs V., Mathur J., Mao R., Bayrak-Toydemir P., Procter M., Cahalan S.M., Kim H.J., Bandell M., Longo N., Day R.W., Stevenson D.A., Patapoutian A, & Krock B.L. (2015). Impaired PIEZO1 function in patients with a novel autosomal recessive congenital lymphatic dysplasia. Nature Communications, 6, 8329.

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TRPV1-dependent Whole-cell Patch Clamp

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TRPV1-dependent currents were examined by whole-cell patch clamp recordings at room temperature in transfected HEK cells. Data were acquired and analyzed using an Axopatch 200B amplifier and pCLAMP10 software (Axon Instruments, Union City, CA, USA). Currents were filtered with a low pass Bessel filter at 1 kHz and sampled at 5 kHz. Borosilicate pipettes (Sutter, Novato, CA, USA) were polished to resistances of 2–5 MΩ. I–V relations were obtained as previously described [8 (link)]. Extracellular bath solution contained (in mM) 135 NaCl, 5 KCl, 2CaCl2, 1 MgCl2, 10 Glucose, 10 HEPES 5 Tris-base, and pH 7.4 with NaOH. Intracellular solutions contained (in mM) 140 KCl 1 MgCl2, 1 EGTA, 5 MgATP, 1 Na-GTP, 10 HEPES, 5 Tris-base, and pH 7.1 with KOH.

DelloStritto D.J., Sinharoy P., Connell P.J., Fahmy J.N., Cappelli H.C., Thodeti C.K., Geldenhuys W.J., Damron D.S, & Bratz I.N. (2016). 4-Hydroxynonenal dependent alteration of TRPV1-mediated coronary microvascular signaling. Free radical biology & medicine, 101, 10-19.

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Whole-cell Calcium Current Recording

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A few drops of cell suspension were placed into a perfusion chamber that was mounted on an inverted microscope (IX70, Olympus, Japan). After the cells adhered to the bottom of the chamber (approximately 10 min), they were infused with oxygen-saturated Tyrode’s buffer (1.2 mL/min) using a pump (DHL-B, Shanghai, China). Borosilicate pipettes (inside diameter of 0.86 mm, outside diameter of 1.5 mm, Sutter, Novato, CA, USA) were constructed using a micropipette puller (P-97; Sutter, Novato, CA, USA) and had a resistance of 4–6 MΩ. Whole-cell currents were recorded with an Axopatch 700B amplifier (Axon Instruments, Burlingame, CA, USA), and the data were digitized at 1 kHz and filtered at 800 Hz. Acquisition and analysis of the physiological signals were carried out using pClamp 10.2 software (Axon Instruments, Burlingame, CA, USA). The experiment was performed at room temperature (approximately 25°C). The pipette solution for recording L-type calcium currents (I_Ca,L) contained 135 mM CsCl, 3 mM MgCl₂, 2 mM Na₂ATP, 10 mM ethylene glycol-bis (2-aminoethyl-ether)-N,N,N’,N’-tetraacetic acid (EGTA), 10 mM HEPES, and 20 mM tetraethylammonium (TEA) (pH adjusted to 7.3 with CsOH).

Ren H., Yuan F., Tan W., Ding Y., An P, & Luo H. (2021). Effect of Evodiamine on Rat Colonic Hypermotility Induced by Water Avoidance Stress and the Underlying Mechanism. Drug Design, Development and Therapy, 15, 441-452.

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Erythrocyte Calcium Imaging Protocols

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Erythrocyte calcium imaging experiments were performed as previously described¹⁵. Briefly, blood was drawn and stored on ice in EGTA-vacutainers. On the experimental day we diluted whole blood 1:1000 into NECM supplemented with 0.1% BSA and loaded with 5 μM Fluo-4 at 4°C for at least an hour. Cells were then washed with buffer to remove excess dye and allowed to settle in a custom-build low volume perfusion chamber, where a gravity-driven whole chamber perfusion system was used to deliver Yoda1 and the calcium ionophore A23187 after erythrocytes loosely adhered to the uncoated glass bottom chamber. For mechanical stimulation we used long, tapered tip (~1.5-2 μm diameter) borosilicate pipettes (Sutter instruments, Novato, CA) forged with a Flaming/Brown P-97 puller (Sutter Instruments) and applied negative pressure to the cells using a High-speed Pressure Clamp device (ALA scientific, Farmingdale, NY). Imaging was performed using an Axio Observer (Zeiss) microscope fitted with a lambda DG4 fluorescent excitation source (Sutter Instruments). Images were recorded with an Orca Flash 4.0 camera (Hamamatsu Photonics) using MicroManager imaging software³³. Image analysis was performed with FIJI^{34 (link)}. HEK-P1KO cells were loaded with 2 μM Fura2-AM for 30 minutes at RT in NECM and imaged using the equipment described above.

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Electrophysiological Characterization of DRG Neurons

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Dorsal root ganglia (DRG) neurons were dissected bilaterally and cultured for electrophysiology. All recordings were made in whole-cell voltage clamp (holding potential (V_h) of –60 mV) configuration at 22–24 °C from the somata of sensory neurons (15–40 pF). Data were acquired and analyzed using Axopatch 200B amplifiers and pCLAMP9.2 software (Molecular Devises, CA). Recording data were filtered at 0.5 kHz and sampled at 2 kHz. Borosilicate pipettes (Sutter, Novato, CA) were polished to resistances of 3–5 M□ in the standard pipette solution. Access resistance (R_s) was not compensated. Data were rejected when R_s changed >20% during recording, leak currents were >100 pA, or input resistance was <200 M. Currents were considered positive when their amplitudes were 5-fold larger than displayed noise (in root mean square). Standard external solution (SES) contained (in mM): 140 NaCl, 5 KCl, 2 CaCl₂, 1 MgCl₂, 10 D-glucose and 10 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), pH 7.4. The pipette solution for the perforated patch configurations consisted of (in mM): 140 KCl, 5 NaCl, 10 EGTA, 1CaCl₂, 1 MgCl₂, and 10 HEPES pH 7.3. Drugs were applied using a fast, pressure-driven and computer controlled 8-channel system (AutoMate Scientific, San Francisco, CA).

Brackley A.D., Gomez R., Guerrero K.A., Akopian A.N., Glucksman M.J., Du J., Carlton S.M, & Jeske N.A. (2017). A-Kinase Anchoring Protein 79/150 Scaffolds Transient Receptor Potential A 1 Phosphorylation and Sensitization by Metabotropic Glutamate Receptor Activation. Scientific Reports, 7, 1842.

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Whole-cell Patch Clamp Recording in Transfected Cells

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The whole-cell patch clamp recordings were performed at room temperature in transfected HEK and BAEC cells. Data were acquired and analyzed using an Axopatch 200B amplifier and pCLAMP10 software (Axon Instruments, Union City, CA, USA). Currents were filtered with a low pass Bessel filter at 1 kHz and sampled at 5 kHz. Borosilicate pipettes (Sutter, Novato, CA, USA) were polished to resistances of 0.5–3 MΩ. I–V relations were obtained as previously described [6 (link)]. After whole-cell access was established, series resistance and membrane capacitance were compensated as completely as possible. Current–voltage relationships were assessed by 400-ms step pulses from −100 to +100 mV in 20-mV increments from −40 mV holding potential. Steady state currents (average of 350–400 ms intervals) were used to generate I/V plots. Similarly, MCECs were held and recorded at −60 mV holding potential in response to H₂O₂. In whole-cell patch the extracellular bath solution contained (in mM): 135 NaCl, 5 KCl, 2CaCl₂, 1 MgCl₂, 10 Glucose, 10 HEPES 5 Tris-base, and pH 7.4 with NaOH. Intracellular solutions contained 140 KCl 1 MgCl₂, 1 EGTA, 5 MgATP, 1 Na-GTP, 10 HEPES, 5 Tris-base, and pH 7.1 with KOH.

DelloStritto D.J., Connell P.J., Dick G.M., Fancher I.S., Klarich B., Fahmy J.N., Kang P.T., Chen Y.R., Damron D.S., Thodeti C.K, & Bratz I.N. (2016). Differential regulation of TRPV1 channels by H2O2: implications for diabetic microvascular dysfunction. Basic research in cardiology, 111(2), 21.

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Whole-Cell Patch Clamp Recording of Cultured Trigeminal Ganglion Neurons

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All I_CAP recordings from the somata of cultured TGs (15–60 pF) were made in the whole cell patch configuration with a holding potential (V_h) of −60 mV to represent physiological resting membrane potential. Data were acquired and analyzed with an Axopatch200B amplifier and pCLAMP 10 software (Molecular Devices, Union City, CA) with recordings filtered at 0.5 kHz and sampled at 2 kHz. Borosilicate pipettes (Sutter Instruments, Novato, CA) were polished to resistances of 5–10 MΩ in pipette solution. Access resistance was compensated (40–80%) when appropriate. SES was the same as for calcium imaging experiments supplemented with 2 mM Ca²⁺. Pipette solution contained SES supplemented with 0.2 mM Na-GTP, 2.5 mM Mg-ATP, and 1 mM CaCl₂ (pH 7.3) as described [30] (link). CAP (100 nM) was locally applied using a computer-controlled application system (MicroData Instrument, South Plainfield, NJ).

Rowan M.P., Bierbower S.M., Eskander M.A., Szteyn K., Por E.D., Gomez R., Veldhuis N., Bunnett N.W, & Jeske N.A. (2014). Activation of Mu Opioid Receptors Sensitizes Transient Receptor Potential Vanilloid Type 1 (TRPV1) via β-Arrestin-2-Mediated Cross-Talk. PLoS ONE, 9(4), e93688.

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Electrophysiological Profiling of NPY Neurons

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Brain slices (250μm) were prepared from adult NPY-GFP mice (6-8 weeks old) as previously described (7) . Slices were incubated at room temperature (RT), in oxygenated extracellular medium containing (in mM): 118 NaCl, 3 KCl, 1 MgCl 2 , 25 NaHCO3, 1.2 NaH 2 PO 4 , 1.5 CaCl 2 , 5 Hepes, 2.5 D-glucose (osmolarity adjusted to 310mOsM with sucrose, pH 7.3) for a recovery period (at least 60minutes). Once in the recording chamber, slices were perfused at 2-3 ml/min with the same extracellular medium. Slices were viewed with a Nikon microscope (EF600) outfitted for fluorescence (fluorescein filter) and IR-DIC (Infrared-Differemcial interference contrast) videomicroscopy. Viable arcuate NPY neurons were visualized using a X60 water immersion objective (Nikon) with a fluorescence video camera (Nikon).
Borosilicate pipettes (4-6MΩ; 1.5mm OD, Sutter Instrument) were filled with filtered extracellular medium. Cell-attached recordings were made using a Multiclamp 700B amplifier, digitized using the Digidata 1440A interface and acquired at 3kHz using pClamp 10.3 software (Axon Instruments). Pipettes and cell capacitances were fully compensated.
After a stable baseline was established, BHB (5mM) was perfused for 10minutes. The firing activity was measured over the last minute of the BHB perfusion and compared with the firing rate measured 1 min before the perfusion.

Carneiro L., Geller S., Fioramonti X., Hébert A., Repond C., Leloup C, & Pellerin L. (2016). Evidence for hypothalamic ketone body sensing: impact on food intake and peripheral metabolic responses in mice. American journal of physiology. Endocrinology and metabolism, 310(2).

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