The electrochemical recordings were carried out by a CHI 760C electrochemical workstation (CH Instrument, Shanghai, China). The monitoring of the current–voltage (I–V) curves was performed by using the scan rate of 50 mVs–1 and the voltage sweeping from −1.0 to 1.0 V. The θ‐nanopipettes employed in the work were fabricated by a P‐2000 laser‐pulling (Sutter Instruments, USA). The θ‐nanopipette is anchored to the holder (Axon Instruments, USA), and further precisely controlled by a MP‐225 micromanipulator (Sutter Instrument, USA) under the observation of Olympus inverted fluorescence microscope (Ti2‐E, Nikon, Japan). Before electrochemical measurements, the tip of the θ‐nanopipette was backfilled by 20 mM Tris‐HCl (pH 7.4) with LiCl of 40 mM and checked under the microscope for the exclusion of the air bubbles. Field‐emission scanning electron microscopic (FE‐SEM) images were collected by a S‐4800 Instrument (Hitachi Co., Japan). Au coating was carried out with the vacuum evaporation equipment (ACE600, Leica, Germany). The liquid‐filled θ‐nanopipettes were vacuumed in the small transition chamber next to the glove box (Vigor, China). Confocal laser scanning microscopy (CLSM) characterization was performed using a Leica TCS SP5 microscope (Germany).
Mp 225 micromanipulator
Manufactured by Sutter Instruments
Sourced in United States
The MP-225 micromanipulator is a precision positioning device designed for delicate laboratory tasks. It provides precise, three-dimensional control for the manipulation of small objects or instruments. The MP-225 features a compact, lightweight design and is suitable for use in a variety of laboratory settings.
Automatically generated - may contain errors
Lab products found in correlation
S 4800 instrument, by Hitachi (1 mentions)
Ti2 e, by Nikon (1 mentions)
Tcs sp5 microscope, by Leica (1 mentions)
Chi 760c electrochemical workstation, by CH Instruments (1 mentions)
Ace600, by Leica (1 mentions)
P 97 pipette puller, by Sutter Instruments (1 mentions)
Pclamp software package, by Molecular Devices (1 mentions)
Axopatch 200b amplifier, by Molecular Devices (1 mentions)
Digidata 1440a, by Molecular Devices (1 mentions)
5 protocols using mp 225 micromanipulator
1
Electrochemical Characterization of Nanopipettes
2
Nanosecond Pulse Delivery for Cell Stimulation
3
Nanosecond pulses for chromaffin cell study
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The 5 ns duration pulses were delivered by a nanosecond pulse generator, designed and fabricated by Transient Plasma Systems, Inc. (Torrance, CA), to chromaffin cells using a pair of cylindrical, gold-plated tungsten rod electrodes (127 μm diameter) with a gap of 100 μm between the electrode tips as detailed in previous publications [6 (link), 13 (link)]. After achieving the whole-cell recording mode by rupturing the cell plasma membrane, the NEP-delivering electrodes were positioned to a predefined “working” position 40 μm from the bottom of the coverslip by a motorized MP-225 micromanipulator (Sutter Instruments, Novato, CA), with the patched cell situated midway between the electrode tips. A single or pair of 5 ns pulses was delivered to the cells at E-field amplitudes ranging from 5 to 10 MV/m. The E-field distribution at the location of the target cell was computed by the Finite-Difference Time-Domain (FDTD) method using the software package SEMCAD X (version 14.8.5, SPEAG, Zurich, Switzerland) as previously described [5 (link), 6 (link), 13 (link)]. In all experiments, two sham (control) exposures preceded the delivery of a single or twin NEP(s) controlled by a program written in LabVIEW [5 (link), 6 (link), 13 (link)].
4
Pulsed Electric Field Delivery to Murine Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The system used for delivering 5 ns electric pulses to murine ACC was the same as that used in our previous studies that employed bovine ACC [42 (link),43 (link)]. Briefly, a single pulse was applied to a cell by means of two cylindrical gold-coated tungsten rod electrodes (diameter of rods 127 μm) in which the electrode tips were spaced 100 μm apart. The electrodes were immersed in the BSS and positioned 40 μm above the bottom of the dish by a motorized MP-225 micromanipulator (Sutter Instruments, Novato, CA), with the cell being exposed to the pulse positioned in the center of the gap between the electrode tips. Pulses 5 ns in duration [42 (link),43 (link)] were generated by a custom-fabricated pulse generator (Transient Plasma Systems, Inc., Torrance CA) and delivered to the electrodes at amplitudes that produced an electric field of 8 MV/m at the location of the cell. Delivery of pulses was triggered externally by a program written in LabVIEW and each pulse trace was captured with an oscilloscope. The electric field distribution in the vicinity and at the location of the target cell was computed using the Finite-Difference Time-Domain software package SEMCAD X (version 14.8.5, SPEAG, Zurich, Switzerland) as previously described [42 (link),43 (link)].
5
Whole-cell patch-clamp electrophysiology protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Electrodes were pulled from glass capillaries with filament (Sutter Instruments) with a P-97 pipette puller (Sutter Instruments) to resistances of 4-7 MΩ. Electrodes were filled with a K-gluconate internal solution (pH 7.25; 285 mOsmol/L, composition in mM: 125 potassium gluconate, 10 KCl, 5 NaCl, 1 EGTA, 10 HEPES, 2 ATP sodium salt, 0.3 GTP sodium salt). EGTA (tetraacid form) was prepared as a stock solution in 1 M KOH before addition to the internal solution. Voltage steps were corrected for the calculated liquid junction (pClamp software package, Molecular Devices) between IB and the K-gluconate internal. 15 (link) Electrodes were position with an MP-225 micromanipulator (Sutter Instruments) to obtain a gigaseal prior to breaking into the whole cell configuration. Recordings were sampled at a rate of > 10 kHz using an Axopatch 200B amplifier, filtered with a 5 kHz low-pass Bessel filter, and digitized with a DigiData 1440A (Molecular Devices). Only recordings that maintained a 30:1 ratio of membrane resistance Rm to access resistance Ra were used for analysis. Pipette capacitance was corrected with the fast magnitude knob only; series resistance compensation was not performed. Voltage steps of -80, -40, 0, and +40 mV were applied in random order, followed by a voltage step to +80 mV.
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
Revolutionizing how scientists
search and build protocols!