Model p 1000

Manufactured by Sutter Instruments

Sourced in United States

The Model P-1000 is a precision micropipette puller manufactured by Sutter Instruments. It is designed to create micropipettes and microelectrodes from glass or quartz capillaries. The device uses heat and precise mechanical force to pull and shape the capillaries into the desired tip diameter and taper.

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Lab products found in correlation

Igor pro, by Wavemetrics (3 mentions) Epc 10 amplifier, by HEKA Elektronik (3 mentions) Axopatch 700b amplifier, by Molecular Devices (2 mentions) Cat no 22102, by AAT Bioquest (2 mentions) Eos rebel t5i camera, by Canon (1 mentions) Nanoliter 2010 microinjector, by World Precision Instruments (1 mentions) Borosilicate glass capillary tubes, by Harvard Apparatus (1 mentions) Multiclamp 700b amplifier, by Molecular Devices (1 mentions) Nanoject 3, by Drummond (1 mentions) Ccd camera c11440, by Hamamatsu Photonics (1 mentions) Axonpatch 700b amplifier, by Molecular Devices (1 mentions) Video camera, by Philips (1 mentions) Eclipse e2000fn microscope, by Nikon (1 mentions) Bf150 110 10, by Sutter Instruments (1 mentions) Axopatch 200a patch clamp amplifier, by Molecular Devices (1 mentions) Matlab, by MathWorks (1 mentions) Originpro, by OriginLab (1 mentions) Bx50wi, by Olympus (1 mentions) Digidata 1440a, by Molecular Devices (1 mentions) Orca r2, by Hamamatsu Photonics (1 mentions)

15 protocols using model p 1000

In vitro and in vivo RNAi Assay Protocol

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Methods for in vitro dsRNA synthesis have been described in our recent paper (Yoon, Shukla, Gong, Mogilicherla, & Palli, 2016 (link)). For in vitro RNAi assay, 25,000 TCA cells/well were seeded in 96 well plates. The equal amount (1 μg/μl) of dsRNA was added to cell and pictures were taken on 6th day after dsRNA treatment. The same amount of luciferase dsRNA was used as a negative control. For in vivo RNAi assay, 200 ng of dsRNA was injected into the dorsal side of the abdomen using a Drummond Nanoject III fitted with 3.5″ glass capillary tube, pulled by a needle puller (Model P-1000, Sutter Instruments). Newly molted last instar larvae were injected. After injection, the insects were reared under standard conditions. Mortality was recorded after 14 days. Those insects showing distinct phenotypes after dsIAP1 treatment were treated with phosphate-buffered saline (PBS) + 0.1% trypan blue solution to identify the excreted substance. The phenotype of the quiescent stage larvae and adults injected with dsTcIAP5 were photographed using a digital microscope system (UNITRON Z850 Stereo Microscope, CB-ZM deep focus extension module, QuickPHOTO industrial 3.1 program, Canon EOS Rebel T5i camera).

Yoon J.S., Koo J., George S, & Palli S.R. (2020). Evaluation of inhibitor of apoptosis genes as targets for RNAi-mediated control of insect pests. Archives of insect biochemistry and physiology, 104(4), e21689.

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Xenopus Oocyte Microinjection and Cryo-sectioning

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Defolliculated stage V and VI oocytes (diameter ~1.2 mm) from Xenopus laevis were obtained from Ecocyte Bioscience in April—May 2019 and stored in ND96 saline (NaCl 96 mM, KCl 20 mM, CaCl₂.2H₂0 1.8 mM, MgCl₂.6H₂0 1 mM, HEPES 5 mM, pH 7.6) supplemented with sodium pyruvate (2.5 mM) and 1 ml/100 ml penicillin-streptomycin (10,000 units penicillin, 10 mg/ml streptomycin). Storage media were changed daily, and oocytes were stored at 17 °C. The oocytes were injected with 18 nL of eGFP cDNA (200 μg ml⁻¹) using a Nanoliter 2010 micro-injector (World Precision Instruments). Injection micropipettes were pulled on a Sutter Instrument Model P-1000, using fire polished glass capillaries (ID = 0.530 mm, OD = 1.14 mm). Prior to injection, the oocytes were aligned with the animal pole facing upward, and the injection needle was positioned for best approximation for direct nuclear injection. Following injection, oocytes were returned to ND96 saline for incubation for 48 h at 17 °C and then dissected by cryo-sectioning.

Zeng S.L., Grabowska D., Shahverdi K., Sudlow L.C., Achilefu S, & Berezin M.Y. (2019). Fluorescence lifetime imaging reveals heterogeneous functional distribution of eGFP expressed in Xenopus oocytes. Methods and applications in fluorescence, 8(1), 015001.

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Spontaneous Firing Monitoring of VLPO Neurons

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Loose cell-attached patch-clamp recordings were performed to monitor the stable spontaneous firing activity of VLPO neurons for long periods of time, as a prerequisite for completing the pharmacological experiments. Infrared videomicroscopy was used to visually select VLPO neurons. Loose-cell attached recordings were performed from the soma with patch micropipettes (3–6 MΩ) pulled from borosilicate glass capillary tubes (1.5 mm o.d., 0.86 mm i.d.; Harvard Apparatus, France) on a horizontal puller (Model P-1000; Sutter Instrument, Novato, CA, USA). Micropipettes were filled with oxygenated aCSF and fixed to an electric Microdrive (Luigs and Neumann, Germany). The micropipette was placed in contact with the soma of a selected neuron under visual control. During recordings, a seal resistance of 10–15 MΩ was maintained to avoid damage or mechanical stimulation to the cell.

Scharbarg E., Daenens M., Lemaître F., Geoffroy H., Guille-Collignon M., Gallopin T, & Rancillac A. (2016). Astrocyte-derived adenosine is central to the hypnogenic effect of glucose. Scientific Reports, 6, 19107.

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Extracellular Recording of Purkinje Neurons

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Extracellular recordings were obtained from PNs in slices constantly perfused with carbogen-bubbled extracellular solution as above with additional compounds as indicated. Cells were visualized with DIC optics and a water-immersion 40× objective (NA 0.75). Glass pipettes of ~3 MΩ resistance (Model P-1000, Sutter instruments, Novato, CA) when filled with extracellular solution were positioned near PN axon hillocks in order to measure action potential-associated capacitive current transients in voltage clamp mode with the pipette potential held at 0 mV. Data was acquired using a Multiclamp 700B amplifier at 20 kHz, Digidata 1440 with pClamp10 (Molecular Devices, Sunnyvale, CA) and filtered at 4 kHz. A total of 20 to 45 PNs were measured from each genotype and each recording was for 10 to 15 min. For each genotype 6 to 8 mice were used and the experimenter was blinded to the genotype.

Meera P., Pulst S, & Otis T. (2017). A positive feedback loop linking enhanced mGluR function and basal calcium in spinocerebellar ataxia type 2. eLife, 6, e26377.

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Microinjection of dsRNA in Insect Larvae

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Newly molted final instar larvae were used for microinjection. 400 ng of dsRNA was injected into each larva on the dorsal side of the abdomen. The NANOJECT III (Drummond Scientific Co.) was used for dsRNA microinjection with an aspirator tube assembly ﬁtted with 3.5″ glass capillary tube (Drummond Scientific Co.) pulled by a needle puller (Model P-1000, Sutter Instruments Co., Novato, CA). Injected larvae were allowed to recover for 1 h at room temperature and then transferred to a 30°C incubator under standard conditions. Control larvae were injected with dsRNA targeting E. coli malE gene.

Luo G.H., Chen X.E., Jiao Y.Y., Zhu G.H., Zhang R., Dhandapani R.K., Fang J.C, & Palli S.R. (2022). SoxC is Required for Ecdysteroid Induction of Neuropeptide Genes During Insect Eclosion. Frontiers in Genetics, 13, 942884.

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Visualizing and Electrophysiologically Characterizing Pyramidal Neurons in vmPFC

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mCherry⁺ (Supplementary Fig. 11) or eYFP⁻ (Fig. 7) pyramidal neurons in layer 2/3 of vmPFC were visualized via infrared oblique-illumination (Olympus X51W; Hamamatsu CCD camera C11440) and randomly selected for voltage-clamp recordings. Patch pipettes (4–6 MΩ) were made from borosciliate glass using a micropipette puller (Model P-1000; Sutter Instruments). These pipettes were filled with pipette solution contained (in mM): 132.5 Cs-gluconate, 17.5 CsCl, 2 MgCl₂, 0.5 EGTA, 10 HEPES, 4 ATP, 5 QX-314 (pH 7.3). Evoked inhibitory post-synaptic currents (eIPSCs) were recorded by voltage clamping at +10 mV^{68 (link)} using an Axonpatch-700B amplifier (Axon Instruments). For optogenetic stimulation of ChR2-eYFP⁺ nNOS-expressing neurons in vmPFC, Single light pulse (5 ms duration) was delivered to the slice through an optical fiber from a 465 nm wavelength blue laser. All recordings were performed in the presence of TTX (0.5 μM) and 4-AP (100 μM) to obtain monosynaptic transmission.

Liang H.Y., Chen Z.J., Xiao H., Lin Y.H., Hu Y.Y., Chang L., Wu H.Y., Wang P., Lu W., Zhu D.Y, & Luo C.X. (2020). nNOS-expressing neurons in the vmPFC transform pPVT-derived chronic pain signals into anxiety behaviors. Nature Communications, 11, 2501.

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Whole-cell Voltage Clamp Recordings of Cones

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Whole-cell voltage clamp recordings were made from cones. The recording chamber containing the isolated retina was mounted on a Nikon Eclipse E2000FN microscope (Nikon, Japan) and viewed with a Nikon 60× water immersion objective with infrared differential interference contrast and a video camera (Philips, Netherlands). Recording electrodes were pulled from borosilicate glass (BF-150-110-10; Sutter Instruments, Novato, CA, USA) with a Flaming/Brown micropipette puller (Model P-1000; Sutter Instruments, Novato, CA, USA). The impedances ranged from 8 MΩ to 12 MΩ when filled with pipette medium and measured in bathing solution. Pipettes were connected to an Axopatch 200A patch clamp amplifier (Molecular Devices, Sunnyvale, CA; four-pole low-pass Bessel filter setting, 2 kHz). Signal software (v. 3.07; Cambridge Electronic Design (CED), Cambridge, UK) was used to generate voltage command outputs and to acquire data. Signal software (v. 3.07; CED), MatLab (v2016b, MathWorks), Igor.pro (WaveMetrics, Portland, OR, USA) and Origin Pro (v8, Origin Lab Corporation), were used to analyze the data. All data shown are corrected for the junction potential.

Cenedese V., de Graaff W., Csikós T., Poovayya M., Zoidl G, & Kamermans M. (2017). Pannexin 1 Is Critically Involved in Feedback from Horizontal Cells to Cones. Frontiers in Molecular Neuroscience, 10, 403.

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Whole-cell Recordings of OXTR-Venus Neurons

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OXTR-Venus neurons in the AVPV were identified with an epifluorescence microscope (Olympus BX50WI, Tokyo) equipped with a 40x water immersion lens (0.8 n.a.) and a CCD camera (ORCA-R², Hamamatsu Photonics, Hamamatsu). Whole-cell membrane potential recordings were obtained with an Axopatch 700B amplifier (Molecular Devices, Foster City, CA). Traces were acquired digitally at 20 kHz and filtered at 5 kHz with a Digidata 1440A and in conjunction with PClamp 10 software (Molecular Devices, Foster City, CA). Patch electrodes were drawn with a Flaming/Brown micropipette puller (Model P-1000, Sutter Instrument Novato, CA) from borosilicate capillary glass tubing (1.1 mm ID, 1.5 mm OD, Sutter Instruments, Novato, CA) to have resistance of 4-8MΩ when filled with a pipette solution that contained (in mM): 140 K-Gluconate, 1 MgCl₂, 10 HEPES, 10 CaCl₂, 2 ATP(Mg⁺⁺), and 0.4 GTP (Na⁺) and 11 EGTA. ACSF was saturated with 95% O₂/5% CO₂ and was warmed to 33–34°C during the recordings. Picrotoxin (100 μM) and 6,7-dinitro-quinoxaline-2,3(1H,4H)-dione (DNQX; 10 μM) were also added to ACSF to block the synaptic activity. The estimated liquid junction potential was +9.2mV; however, the data presented are not corrected for liquid junction potential.

Sharma K., LeBlanc R., Haque M., Nishimori K., Reid M.M, & Teruyama R. (2019). Sexually dimorphic oxytocin receptor-expressing neurons in the preoptic area of the mouse brain. PLoS ONE, 14(7), e0219784.

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Whole-cell Voltage-clamp Recordings of dLGN Neurons

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Whole-cell voltage-clamp recordings of dLGN neuron were performed using an EPC-10 amplifier (HEKA Elektronik) controlled by Patchmaster software. Recordings were carried out using a cesium methanesulfonate pipette solution contained (in mM): 100 CsCH₃SO₃, 20 KCl, 7 Na₂-phosphocreatine, 10 HEPES, 4 Mg-ATP, 0.3 GTP, and 10 EGTA, pH adjusted to 7.3 with KOH. The holding potential was −60 mV for AMPAR-mediated current and +40 mV for NMDAR-mediated current recordings in the voltage-clamp mode. Pipettes were pulled using an electrode puller (Model P-1000, Sutter Instruments) to open tip resistances of 4−5 MΩ. dLGN neurons and optic tract were identified based on neuronal tracer Dil (AAT Bioquest, Cat no. 22102, 2% w/v in DMSO) injected into the retina at P9²⁷. A bipolar electrode (Frederic Haer, Bowdoinham, ME) was placed in the optic tract then neurons in the dLGN were recorded. An Iso-Flex stimulator driven by a Master 10 pulse (<10 V constant voltage) was used. Data were analyzed offline and displayed with Igor Pro (Wavemetrics, Lake Oswego, OR).

Cong Q., Soteros B.M., Wollet M., Kim J.H, & Sia G.M. (2020). The endogenous neuronal complement inhibitor SRPX2 protects against complement-mediated synapse elimination during development. Nature neuroscience, 23(9), 1067-1078.

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Whole-cell Voltage-clamp Recordings of dLGN Neurons

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