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S48 stimulator

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The S48 stimulator is a versatile laboratory device designed for a range of applications. It provides electrical stimulation to elicit physiological responses, supporting various experimental and research needs.

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23 protocols using s48 stimulator

1

Optogenetic Stimulation of DRG Neurons

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VGAT-ChR2-EYFP mice were from The Jackson Laboratory. The implantation of optical fiber was performed similarly to the DRG cannula implant with modifications. A stainless steel cannula guide (RWD Life Science Co. Ltd., China; diameter 0.64 mm) was forced into the hole in the transverse process over the L5 DRG, and the optical fiber (RWD Life Science Co. Ltd., China; diameter 0.2 mm, length 1 m) was inserted through the guide. The incision was closed with sutures, and the fiber was firmly fixed in place with dental cement; the rest of the procedure was similar to the cannula implantation. Laser stimulation (473 nm, 3 mW, 30 Hz for 10 seconds with 20-second interval) was elicited using an MLL-FN-473-50 unit (Changchun New Industries Optoelectronics Technology Co., Ltd.) controlled by a pulsing set (S48 Stimulator, Grass Technologies, An Astro-Med, Inc. Product Group).
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2

Penile Erection Monitoring via ICP Measurement

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For the continuous ICP monitoring, the shaft of the penis was denuded of skin and fascia, and the right crus was punctured with a 23-gauge needle connected via polyethylene-50 tubing to a pressure transducer. For electrically stimulated penile erections, a bipolar electrode attached to a Grass Instruments S48 stimulator (Quincy, MA, USA) was placed on CNs. Supramaximal stimulation was induced for 1 minute with 4 volts, 16 Hz, and 5-msec square-wave duration22 (link). ICP was recorded (DI-190, Dataq Instruments, Akron,OH, USA) for 5 minutes; pre-stimulation (1 minute), stimulation (1 minute), and post-stimulation (3 minutes). Results were analyzed using the MATLAB program (Mathworks, Natick, MA, USA). At the conclusion of experiments, animals were sacrificed by a lethal intracardiac injection of saturated potassium chloride and prostate, both MPGs, and CNs were collected.
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3

Measuring Intracranial Pressure Response to Electrical Stimulation

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ICP was measured in anesthetized (100 mg/kg ketamine/5 mg/kg xylazine) animals [27 (link)]. The electrode was attached to a Grass Instruments S48 stimulator. The stimulation parameters were 16Hz at 1V and 4V with monophasic, square-wave duration of 5ms. The duration of electrical stimulation of CN was 1 min. Response parameters were recorded using data acquisition (DI-190, Dataq Instruments, Akron, OH, USA), and results were analyzed using Matlab software (Mathworks, Natick, MA, USA). Maximum ICP (maximum pressure that is reached during CN electrical stimulation) and total ICP (ICP area under the curve, indicating the ICP response for the duration of CN electrical stimulation) were expressed per mean arterial pressures (MAP) as a difference from baseline.
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4

GCaMP Imaging and Odor Stimulation in Drosophila

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GCaMP imaging was performed as previously described (Wang et al., 2003 (link); Root et al., 2008 (link)). In odor experiments, a constant airflow of 1 l/min was applied to the antennae via a pipe of 12 mm diameter. Odor onset was controlled by mixing a defined percentage of carrier air with air redirected through odor bottles as previously described (Root et al., 2008 (link); Semmelhack and Wang, 2009 (link)). Nerve stimulation was performed with a glass suction electrode and an S48 stimulator (Grass, Warwick, RI) as previously described (Wang et al., 2003 (link); Root et al., 2008 (link)). Stimulation was 1 ms in duration, 10 V in amplitude, and 16 pulses (Figure 4A) and 45 pulses (Figure 4B) at 100 Hz. Starved flies were starved with water for 16–24 hr.
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5

Measuring Erectile Function in Mice

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Mice were anesthetized with 100 mg/kg Ketamine + 10 mg/kg Xylazine by intraperitoneal injection. To monitor mean arterial pressure (MAP), the right carotid artery was cannulated with polyethylene (PE) tubing filled with heparinized saline (100 U/ml). To monitor changes in intracavernosal pressure (ICP), the penis was denuded of skin and fascia and a 30-gauge needle connected via PE tubing to a pressure transducer was inserted into the right crus. The cavernous nerve was affixed with a bipolar electrode attached to a Grass Instruments S48 stimulator (Quincy, MA, USA) and stimulated at 1, 2, and 4 volts at 16 Hz with a 5 millisecond square-wave duration for 1 minute [10 (link)]. ICP was recorded using the DI-190 system (Dataq Instruments, Akron, OH, USA) from the start of electrical stimulation until 60 seconds after stimulation ended. Erectile function was represented by the normalized maximal ICP/MAP (max ICP) and total area under the curve/MAP (total ICP). Results were analyzed using the MATLAB program (Mathworks, Natick, MA, USA).
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6

Electroretinography in Zebrafish Larvae

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Larvae were dark adapted for 30 minutes and paralysed with 0.5 mg/mL mivacurium chloride (Mivacron), the dark-adapted status was maintained throughout the handling and measurement processes. The reference electrode was placed in E3 medium, the recording electrode was filled with 0.9% saline solution and positioned on the center of the cornea using a micromanipulator and amplified with a P55 pre-amplifier (Grass Instruments). A 300 W halogen light source was used for light stimulation. The maximum light intensity stimulus was 2.8 × 103 µW/cm2 with a 20 ms flash duration at all intensities (controlled by a S48 stimulator (Grass Instruments). Three optical density filters produced flash intensities at −3.0, −2.0, −1.0 and −0 log of the maximum. Recordings were analysed as previously described68 (link). Raw data from dye mutants and siblings was compared using a 2-sample t-test with unequal variances.
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7

Electrical Stimulation for Muscle Regeneration

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iNMES was initiated on day 5 after initial transplantation. Mice were anesthetized with 2–2.5 % isoflurane. The ankle dorsiflexors of the injected hindlimb were stimulated electrically through the intact skin over the common peroneal nerve, with the electrode placed over the head of the fibula. Monophasic square wave pulses, 0.1 ms in duration, were delivered to the stimulation electrode by an S48 Stimulator (Grass Instruments, Warwick, RI). A stimulation isolation unit (model PSIU6; Grass Instrument) was used to minimize artifact and to ensure that the peak current delivered was no greater than 15 mA. The iNMES training protocol consisted of four sets of ten contractions. Each contraction lasted 500 ms (150 Hz pulse frequency), followed by a 500-ms rest. Rest times of 2 min were allowed between each set of contractions to minimize the effect of fatigue. iNMES training was performed three times/week for 4 weeks. Most samples were analyzed between 4 and 5 weeks after injection of hMPCs.
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8

Controlling Factors Affecting Blood Flow

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Control experiments were performed to rule out other potential sources of electric charges affecting blood flow measurements. The acute flare reaction from FUS was compared with those obtained from three control experiments: (1) deactivating the FUS driving system without changing the rest of the experimental setup (n=7 hindlimbs of mice), (2) applying 20 dB attenuated FUS with different driving conditions (driving the transducer outside its frequency bandwidth (center frequency of 3.1 Hz with PRF of 0.1 Hz)) without modifying the rest of the experimental setup (n=6 hindlimbs of mice) to investigate the effect of potential sources of electromagnetic noise on the blood flow measurement while activating FUS driving system, and (3) applying off-target FUS at 2.5 mm away from the nerve with the same experimental setup (n=6 hindlimbs of mice). In addition, FUS with a total sonication time of 3 min (n=2 hindlimbs of mice) and electrical stimulation (S48 stimulator, Grass Technologies, West Warwick, RI) with custom-built electrical stimulation wire were separately conducted. For electrical stimulation, we used 50% of the electrical stimulation voltage (2–3V), where 100% of the electrical stimulation voltage (4–6V) was capable of evoking muscle activation in each experiment (n=2 hindlimbs of mice) to compare FUS-induced induced flare response.
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9

Isometric Force Measurement of Muscle Strips

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A muscle strip, including the tendinous attachments at the central tendon and rib cage was dissected from the midcostal region. The strip was suspended vertically between two lightweight Plexiglas clamps with one end connected to an isometric force transducer (model FT-03, Grass Instruments, Quincy, MA) within a jacketed tissue bath. The force output was recorded via a computerized data-acquisition system (Super Scope II, GW Instruments Somerville, MA). The muscle strip was stimulated along its entire length with platinum wire electrodes (modified S48 stimulator, Grass Instruments) by using supramaximal (~150%) stimulation voltage to determine the optimum contractile length (Lo). To measure maximal isometric twitch force each strip was stimulated supramaximally with 120-V pulses at 1 Hz and to measure the force frequency response each strip was stimulated supramaximally with 120-V pulses at 15– 160 Hz [32 (link)].
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10

Olfactory Learning in Drosophila

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Olfactory learning was assayed using olfactory classical conditioning procedures50 (link). All behavioral experiments were performed under a dim red light at 26 °C and 80% relative humidity. Groups of 50–60 flies were transferred to small plastic tubes with a copper-grid floor to deliver the electric shock. A single cycle of training consisted of one presentation of (CS+) for 60 sec along with 90 V, 1.25 sec shock pulses (GRASS S48 Stimulator) every 5 sec, followed by the second odor presentation without associated shock (CS−) for another 60 sec. Odor presentations were separated by 30 sec of fresh air. For each N, two groups of flies of the same genotype were trained and tested simultaneously with the CS+ and CS− odors reversed. Benzaldehyde and 3-octanol were selected as the odor pairs.
After training, the animals were tested immediately in a runway in which they chose between avoiding the CS+ or the CS− odor. Performance index (PI) was calculated by subtracting the number of flies avoiding the CS− odor from the number of flies avoiding the CS+ odor, divided by the total number of flies, and averaged for the two reciprocal half experiments with reversed odors.
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