Micropipette puller

Manufactured by Sutter Instruments

Sourced in United States

The Micropipette Puller is a laboratory instrument used to create glass micropipettes from capillary tubes. The device applies heat and mechanical force to the tube, causing it to pull apart and form two tapered micropipettes.

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Lab products found in correlation

Inverted microscope, by Nikon (3 mentions) Pclamp 10, by Molecular Devices (3 mentions) Ms 222, by Merck Group (2 mentions) M 152 micromanipulator, by Narishige (2 mentions) Bf100 50 10, by Sutter Instruments (2 mentions) Picopump pv820, by World Precision Instruments (2 mentions) Ix2 slp, by Olympus (2 mentions) Matrigel coated glass coverslips, by Warner Instruments (2 mentions) Rc 26c recording chamber, by Warner Instruments (2 mentions) Ultima gold scintillation cocktail, by PerkinElmer (2 mentions) E600fn, by Nikon (2 mentions) Multiclamp 700b, by Molecular Devices (2 mentions) Digidata 1322a, by Molecular Devices (2 mentions) Inverted microscope, by Eppendorf (2 mentions) Femtojet 4i injector, by Eppendorf (2 mentions) Vacutip 1, by Eppendorf (2 mentions) Trans ferman nk2 micro manipulators, by Eppendorf (2 mentions) Amphotericin b, by Merck Group (2 mentions) Patch chamber, by Warner Instruments (2 mentions) Filamented glass capillary tubes, by World Precision Instruments (2 mentions)

47 protocols using micropipette puller

Intracerebral Aβ Injections in Larval Zebrafish

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Aβ injections were carried out with a Pneumatic PicoSpritzer III (Parker Instrumentation) with glass capillary needles (Harvard Apparatus # GC100F-15) prepared with a Micropipette Puller (Sutter Instruments, # P-87) HiLyte647-conjugated Aβ1-40 (Anaspec) was constituted in PBS, diluted to 1 mg/mL and kept on ice until the point of injection. The injected embryos were anaesthetized in 2% low-melting agarose (ThermoFischer, #16520100) dissolved in embryo medium containing 42 mg/L MS222 (Sigma, #A5040) and injected with a total volume of 1 nL per injected bolus. Needles were inserted into the brain in a sloped angle into the CSF directly dorsal lateral to the posterior region of the hindbrain ventricle.
Live larval fish and whole mounts were imaged with a Leica SP8 laser scanning confocal microscope (LSCM) using 25× and 40× water immersion objectives.

Shibata-Germanos S., Goodman J.R., Grieg A., Trivedi C.A., Benson B.C., Foti S.C., Faro A., Castellan R.F., Correra R.M., Barber M., Ruhrberg C., Weller R.O., Lashley T., Iliff J.J., Hawkins T.A, & Rihel J. (2019). Structural and functional conservation of non-lumenized lymphatic endothelial cells in the mammalian leptomeninges. Acta Neuropathologica, 139(2), 383-401.

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Intracellular Recordings from Neuronal Cells

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Intracellular recordings were made using techniques described previously (Voytenko and Galazyuk, 2008 (link)). Animals were placed inside a single-walled acoustic chamber (Industrial Acoustics), and positioned on an air table 7″ from a freefield speaker. Sharp microelectrodes were pulled from 1.2-mm-diameter quartz glass (Sutter Instruments) on a Flaming-Brown micropipette puller and filled with 3M potassium acetate. Impedance ranged between 40 and 90 MΩ. After placement on the dorsal surface of the brain, the exposure was filled with 4% agar and the electrode was advanced in 3-μm steps using a precision microdrive (Kopf, Model 660). Intracellular responses were amplified (Cygnus Technologies NeuroData IR183A), monitored with Pulse software (v. 8.65) and digitized using a data acquisition system (Heka model EPC-10) at a sampling rate of 100 kHz.

Gay J.D., Voytenko S.V., Galazyuk A.V, & Rosen M.J. (2014). Developmental hearing loss impairs signal detection in noise: putative central mechanisms. Frontiers in Systems Neuroscience, 8, 162.

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Extracellular Potassium Measurements in Transgenic Notch3 Mice

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[K⁺]_e were measured in 19-week-old male WT and TgNotch3^R169C mice using K⁺-selective electrodes prepared as previously described with some modifications (54 (link)). Double-barreled capillaries were pulled with a micropipette puller (Sutter Instruments). The ion-sensitive barrel was filled with potassium chloride (100 mM, in 154 mM NaCl), and the tip (2–3 μM) was silanized and filled with a K⁺-sensitive resin (Liquid Ion Exchanger IE190, WPI). The reference barrel was filled with sodium chloride (154 mM). Before each experiment the K⁺-sensitive microelectrodes were calibrated in K⁺ solution. Three consecutive SDs were induced every 15 minutes by topical application of 1 mm cotton ball soaked in 300 mM KCl onto the right occipital cortex. Measurements of [K⁺]_e were carried out in somatosensory cortex (2 mm lateral and 2 mm caudal to Bregma), more than 2 mm away from the SD induction site, at a depth of 300 μm.

Oka F., Lee J.H., Yuzawa I., Li M., von Bornstaedt D., Eikermann-Haerter K., Qin T., Chung D.Y., Sadeghian H., Seidel J.L., Imai T., Vuralli D., Platt R.M., Nelson M.T., Joutel A., Sakadzic S, & Ayata C. (2022). CADASIL mutations sensitize the brain to ischemia via spreading depolarizations and abnormal extracellular potassium homeostasis. The Journal of Clinical Investigation, 132(8), e149759.

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Microinjection of Fluorescent Amyloid-β in Zebrafish

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Fluorescence (HiLyte Fluor 488, 555, 647)-labeled amyloid-β (1-42) (DAEFRHDSGYEVHHQKLVFFAEDVGSNKGAIIGLMVGGVVIA) peptides were purchased from AnaSpec (Fremont, CA, USA). Aβ was diluted with DMSO at 2 μg μL⁻¹ and stored at −80 °C deep freezer before use. HiLyte-conjugated Aβ (1-42) stock solution was diluted in 1XPBS (phosphate-buffered saline) (1:9 v/v) and the oligomeric form of Aβ was prepared by incubating at 37 °C for 3 days as previously described [20 ]. The monomeric form of Aβ was immediately used without incubation. Microinjections were carried out with a Pneumatic PicoPump (World Precision Instruments, Sarasota, FL, USA) and capillary needles prepared by a Micropipette puller (Sutter instrument, Novato, CA, USA). For ventricle injection, 3 dpf larvae were anesthetized with tricaine (Sigma, St. Louis, MO, USA) and placed in 1% low-melting agarose confocal dish and injected with the total volume of 1–2 nL of the Aβ solution. Trimmed needles were inserted into the ventricular space between the optic tectum and hindbrain, in order to not penetrate into deep brain tissues.

Jeong Y.M., Lee J.G., Cho H.J., Lee W.S., Jeong J, & Lee J.S. (2021). Differential Clearance of Aβ Species from the Brain by Brain Lymphatic Endothelial Cells in Zebrafish. International Journal of Molecular Sciences, 22(21), 11883.

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DIW Glutamate Biosensor Fabrication

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Fig. 1 shows the construction of our DIW glutamate biosensor. A three axis automated microfluidic dispensing system (Pro-EV3 and Ultimus V, Nordson EFD, East Providence, RI) was used as the DIW platform that can position the dispensing tip with ±8 μm accuracy within the working space of 400 mm². A pressurize 3 cc syringe barrel (Nordson EFD, East Providence, RI) was used as the ink reservoir. A custom glass capillary pipette with 30 μm-diameter tip was fabricated using micropipette puller (Sutter Instrument, Novato, CA) to dispense the ink. Here, an output pressure ranged from 10 to 40 psi, and the printing speed was varied from 1 to 5 mm/s. The biosensors were printed on either PDMS or LCP (Ultralam 3850, Rogers Corporation, Chandler, AZ, USA) substrate. PDMS was prepared by spin coating PDMS on a glass slide. The glass slide was precoated with 1 μm layer of Parylene C to promote device release. The biosensors were also printed on a 100-μm-thick LCP sheet. To complete the glutamate biosensor with good selectivity, Nafion and glutamate oxidase were deposited using a previously described method [18 (link)]. After the enzyme immobilization, the samples were stored in room temperature for 48–72 h, and in 4 °C until testing.

Nguyen T.N., Nolan J.K., Cheng X., Park H., Wang Y., Lam S., Lee H., Kim S.J., Shi R., Chubykin A.A, & Lee H. (2020). Fabrication and ex vivo evaluation of activated carbon–Pt microparticle based glutamate biosensor. Journal of electroanalytical chemistry (Lausanne, Switzerland), 866, 114136.

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Measuring Intercellular Spaces in Follicles

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Fluorescein isothiocyanate (FITC)-dextran (Sigma-Aldrich) of 40-kDa molecular weight was used as an intercellular space indicator that binds to membrane molecules for several hours once it has leaked into junction spaces. Multilayered secondary follicles (200–250 μm in diameter) were mechanically isolated from the ovaries of 4-week-old female WT and AQP8^-/- mice (n = 6 for each genotype). The glass injection needle at a diameter of 20 μm used for the FITC-dextran injection was pulled by the micropipette puller (Sutter, Novato, United States). The single-follicle injection manipulation was carried out in a microinjector device (Narishige, Tokyo, Japan). Two microliters FITC-dextran (10 μM) was pipetted into the middle of follicles. Then, several slices of each follicle in the middle portion were quickly imaged using a confocal LSM 710 microscope (Carl Zeiss, Hertfordshire, United Kingdom). The relative estimation of intercellular space of GCs was quantified by mean FITC fluorescent intensity of two slices from each follicle using ImageJ software. Results were expressed as fluorescence intensity per μm². N = 20 follicles for each genotype, and the experiments were repeated in duplicate.

Wang D., Di X., Wang J., Li M., Zhang D., Hou Y., Hu J., Zhang G., Zhang H., Sun M., Meng X., Sun B., Jiang C., Ma T, & Su W. (2018). Increased Formation of Follicular Antrum in Aquaporin-8-Deficient Mice Is Due to Defective Proliferation and Migration, and Not Steroidogenesis of Granulosa Cells. Frontiers in Physiology, 9, 1193.

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Microfluidic Polymersome Synthesis

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Microfluidic devices were fabricated following procedures outlined by Weitz et al. (Ho et al. 2008 (link)) Briefly, two glass capillaries (O.D. 1 mm) were tapered to diameters of approximately 20 and 200 µm using a Flaming/Browning micropipette puller (Sutter P-97). The tip of the smaller (injection) capillary was coated with n-octadecyltrimethoxysilane (Sigma-Aldrich) while the tip of the larger (collection) capillary was coated with 2-[methoxy(polyethyleneoxy)propyl]trimethoxysilane (Gelest, Inc.). The capillaries were coaxially aligned within a square glass capillary tube (O.D. 1.05 mm) with an intercapillary distance of 100 µm. Three 20-gauge syringe tips were epoxied over the square capillary termini to allow injection of each liquid phase with syringe pumps via polyethylene tubing (O.D. 1.32 mm). For polymersome synthesis, three liquid phases were prepared: An aqueous solution comprised of Lysogeny broth (LB, Becton-Dickson), an organic polymer solution containing 2.5 mg/mL of diblock copolymer poly(ethylene glycol)-b-poly(D,L-lactic acid) (mPEG-PDLLA, 5000 MW mPEG and 50,000 MW PDLLA, Polyscitech) and 1.0 mg/mL of poly(lactic acid) (PLA, 15,000 MW, Polysciences) dissolved in a 2:1 ratio of toluene : chloroform, and an aqueous solution comprised of 10 wt% poly(vinyl alcohol) (PVA, 13,000–23,000 MW, 98% hydrolyzed, Sigma-Aldrich) dissolved by heating in deionized water.

Barlow J., Gozzi K., Kelley C.P., Geilich B.M., Webster T.J., Chai Y., Sridhar S, & van de Ven A.L. (2016). High throughput microencapsulation of B. subtilis in semi-permeable biodegradable polymersomes for selenium remediation. Applied microbiology and biotechnology, 101(1), 455-464.

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Hippocampal Long-Term Potentiation Induction

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Brain coronal slices were prepared from these types of mice with the dark or light treatment similar to behavioral protocols during the night by using protocols describe previously39 . Brains were dissected in ice-cold artificial cerebrospinal fluid (ACSF) and cut (400-μm-thick) with a Leica VT 1000 vibratome (Leica Biosystems) at 0 °C. Before being transferred to a submerged recording chamber, slices were recovered for at least 30 min in oxygenated (95% O₂ and 5% CO₂) warm (37 °C) ACSF containing (in mmol) 120 NaCl, 2.5 KCl, 2 CaCl₂, 2 MgSO₄, 26 NaHCO₃, 1.25 NaH₂PO₄, 10 glucose. Hippocampal CA1 LTP experiment were recording at room temperature, and was induced by three trains of HFS (high frequency stimulation, each with 1 pulse at 100 hertz for 1 s) delivered at 30 s intervals. Recording electrodes (4–6 MΩ) were pulled on a Micropipette Puller (Sutter Instruments, California, USA).
LTP was measured by using the averaged amplitude of the fEPSP during the last 10 min recordings39 . Data were acquired by using a Multiclamp 700 B amplifier connected to a Digidata 1440 analog of the digital converter. All recordings were digitized and analyzed using pClamp 10.0 software (Axon Instruments Inc., California, USA).

Shan L.L., Guo H., Song N.N., Jia Z.P., Hu X.T., Huang J.F., Ding Y.Q., Richter-Levine G., Zhou Q.X, & Xu L. (2015). Light exposure before learning improves memory consolidation at night. Scientific Reports, 5, 15578.

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Microinjection of DNA into Zebrafish Embryos

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DNA samples were microinjected into Danio rerio embryos at the first cleavage 20 min after fertilization using an M-152 micromanipulator (Narishige, Japan) and an air-pressure injector PicoPump PV820 (World Precision Instruments, United States) under an inverted microscope Olympus IX2-SLP (Olympus, Japan). The samples were injected into the yolk under the formed germinal disc at an angle of 45° to the surface with the embryo to maximize sample delivery into the yolk center.
The capillaries used with the outer diameter of 20 µm were pulled from glass capillaries (BF100-50–10, Sutter Instrument, United States) by a Micropipette puller (Sutter Instrument, United States). A 1 nl sample was injected into an embryo within 2.8 × 100 ms.
Vectors DNA were isolated from transformed Escherichia coli TG1 using a Plasmid Miniprep kit (Evrogen, Russia). DNA concentration was determined by spectrophotometry using the extinction coefficient of 0.02 ml/(µg × cm) for double-stranded DNA²⁷. The obtained DNA was dissolved in PBS with 0.05% phenol red (Sigma-Aldrich, United Kingdom).

Selina P.I., Karaseva M.A., Komissarov A.A., Safina D.R., Lunina N.A., Roschina M.P., Sverdlov E.D., Demidyuk I.V, & Kostrov S.V. (2021). Embryotoxic activity of 3C protease of human hepatitis A virus in developing Danio rerio embryos. Scientific Reports, 11, 18196.

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Microinjecting Ticks with Powassan Virus

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Adult female H. longicornis ticks were microinjected with POWV II via the anal pore as previously described (Talactac et al., 2017 (link)). Ticks chosen for microinjection were from the same generation and same time post-molt (i.e. 6 weeks post-molt from nymphal stage to adult). Infection via microinjection was achieved by injecting 475 nL of virus stock into the immobilized tick’s anal aperture using glass microneedles, a digitally-operated microinjector with a footswitch, and a dissecting microscope. The volume of virus injected contained approximately 300 focus-forming units (FFU) of POWV II. For the mock-infected groups, an equivalent volume of DMEM media was used. Glass microneedles were made using a micropipette puller (Sutter Instrument) and glass capillaries (World Precision Instruments). The capillaries an internal diameter of 0.530 mm and an outer diameter of 1.14 mm; these micropipettes were pulled such that the tip had a smaller diameter than that of the tick anal pore. Ticks were housed in ACL-3 facilities and were monitored for mortality twice daily for 4 days after microinjection.

Raney W.R., Herslebs E.J., Langohr I.M., Stone M.C, & Hermance M.E. (2022). Horizontal and Vertical Transmission of Powassan Virus by the Invasive Asian Longhorned Tick, Haemaphysalis longicornis, Under Laboratory Conditions. Frontiers in Cellular and Infection Microbiology, 12, 923914.

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