Digidata 1550a

Manufactured by Molecular Devices

Sourced in United States

The Digidata 1550A is a data acquisition system designed for electrophysiology experiments. It provides high-quality digital data acquisition and signal conditioning capabilities.

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Close spelling variants of this product

Axon Digidata 1550A (12 citations) Digidata 1550A digitizer (7 citations) Digidata 1550A data acquisition system (3 citations)

48 protocols using digidata 1550a

Cardiac Action Potential Recording Protocol

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To avoid the potential effects of exogenous Ca buffering, APs were recorded using pipette solutions without Ca chelating agents such as EGTA unless otherwise indicated. Cells were superfused with Tyrode solution. The glass pipette had access resistance of 4–6 MΩ after filling with the internal pipette solution containing (in mmol/L) 110 KCl, 5 NaCl, 5 Mg-ATP, 10 HEPES, 5 phosphocreatine, pH adjusted to 7.2 with KOH. APs were recorded using Axopatch 200B, Digidata 1550A and pClamp 10.5 software (Axon Instruments, Foster City, CA, USA) for data amplification and acquisition. A 2-ms current pulse at 20% above threshold was provided to evoke APs at a cycle length of 2 s (0.5 Hz). Electrophysiological data were analyzed using Clampfit 10.5 and were prepared by Prism. EMD57033 and blebbistatin were kept in DMSO as 3 mmol/L stock solution, and the final concentration of DMSO in experiment solutions was adjusted to 0.1%. CMs were pretreated with 3 μmol/L EMD57033 or blebbistatin for 1 min and then APs measured using pipette solution without EGTA. To measure the APs under excess cytosolic Ca buffering, 14 mmol/L EGTA was added into pipette solution. The AP triangulation index was defined as the APD(90–30)/APD90 ratio. The beat-to-beat instability index of APs was obtained from 14 consecutive APs per cell by calculating the ratio of interquartile range to median APD50 [18 (link)].

Wang L., Kim K., Parikh S., Cadar A.G., Bersell K.R., He H., Pinto J.R., Kryshtal D.O, & Knollmann B.C. (2017). Hypertrophic cardiomyopathy-linked mutation in Troponin T causes myofibrillar disarray and pro-arrhythmic action potential changes in human iPSC cardiomyocytes. Journal of molecular and cellular cardiology, 114, 320-327.

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Ion Channel Electrophysiology Protocol

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The cells were bathed in external solution containing (mM): NaCl 135, KCl 5.4, MgCl₂ 1.0, CaCl₂ 1.8, NaH₂PO₄ 0.33, glucose 5, and HEPES 10 and was adjusted to pH 7.4 with Tris buffer. The pipetted solution contained (in mM): Mg-ATP 3, CsCl 140, HEPES 10 and EGTA 10 and was adjusted to pH 7.2 with Tris buffer. Currents or potentials were amplified using an Axopatch 200B (Axon Instruments) and digitized with a 16-bit analog to digital converter (Digidata 1550A; Axon Instruments). The data were filtered at 5 kHz and was displayed on a computer monitor. Results were analyzed using pClamp software (version 9.2; Axon Instruments) and GraphPad Prism software. All experiments were performed at 30 °C.

Ju S., Lim L., Jiao H.Y., Choi S., Jun J.Y., Ki Y.J., Choi D.H., Lee J.Y, & Song H. (2020). Oxygenated polycyclic aromatic hydrocarbons from ambient particulate matter induce electrophysiological instability in cardiomyocytes. Particle and Fibre Toxicology, 17, 25.

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TRPM8 Whole-Cell Cation Current Recording

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For electrophysiological experiments, whole-cell cation currents mediated by TRPM8 (I_TRPM8) were recorded by whole-cell patch-clamp technologies with an Axon MultiClamp 700B amplifier using the Digidata1550A digitizer (Axon Instruments, Sunnyvale, CA) as described previously [10 (link)]. Briefly, the indicated expression constructs pEGFP-N1 and Flag-TRPM8 were transfected into HEK293T cells. After 48 h of transfection, we selected the cells expressing an approximately equal amount of GFP to record I_TRPM8 at RT in an extracellular solution containing (mM) 150 NaCl, 6 CsCl, 1 MgCl₂, 1.5 CaCl₂, 10 glucose, 10 mM HEPES, pH 7.4 with NaOH. The peptides were filled with pipette solution ((mM): 150 NaCl, 3 MgCl₂, 5 EGTA, 10 HEPES, pH 7.2 with NaOH) to form a tip resistance of 2~4 MΩ. Series resistance was compensated by 75–85% to reduce voltage errors. The holding potential was −60 mV, and details of each pulse protocol are given schematically in the related figures. The densities of the whole-cell TRPM8 current (I_TRPM8) were normalized to the cell capacitance (pA/pF). The data were analyzed using a combination of Clampfit version 11.0 (Molecular Devices), Microsoft Excel, and GraphPad Prism 5 (GraphPad Software Inc., San Diego, CA, USA).

Huang Y., Li S., Liu Q., Wang Z., Li S., Liu L., Zhao W., Wang K., Zhang R., Wang L., Wang M., William Ali D., Michalak M., Chen X.Z., Zhou C, & Tang J. (2022). The LCK-14-3-3ζ-TRPM8 axis regulates TRPM8 function/assembly and promotes pancreatic cancer malignancy. Cell Death & Disease, 13(6), 524.

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Enteroendocrine Cell-Nodose Neuron Coculture

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Enteroendocrine cells and nodose neurons were cocultured as described above. Recordings were carried out at room temperature using a MultiClamp 700B amplifier (Axon Instruments), digitized using a Digidata 1550A (Axon Instruments) interface, and pClamp software (Axon Instruments) for data acquisition. Recordings were made using borosilicate glass pipettes pulled to ~3.5 MW resistance. Extracellular solution contained (in mM): 140 NaCl, 5 KCl, 2 CaCl2,2 MgCl2, 10 HEPES, pH 7.4 (300 to 305 mosmol). For voltage-clamp recordings, intracellular solution contained (in mM): 140 CsF, 10 NaCl, 0.1 CaCl2, 2 MgCl2,1.1 EGTA, 10 HEPES, 10 sucrose (pH 7.25, 290 to 295 mosmol). For current-clamp recordings, intracellular solution contained (in mM): 140 KCl, 0.5 EGTA, 5 HEPES, 3 Mg-ATP, 10 sucrose (pH 7.25, 290 to 295 mosmol). Data are presented as the mean ± SEM, and significance was determined using a two-tailed Student’s t test.

Kaelberer M.M., Buchanan K.L., Klein M.E., Barth B.B., Montoya M.M., Shen X, & Bohórquez D.V. (2018). A gut-brain neural circuit for nutrient sensory transduction. Science (New York, N.Y.), 361(6408), eaat5236.

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Multimodal Physiological Monitoring in Rodents

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ECG and respiratory rate were acquired at 1 kHz and 250 Hz, respectively, using a small animal physiological monitoring device (Harvard Apparatus). Arterial blood pressure was measured through an arterial catheter placed in the femoral artery and connected to a pressure transducer and monitor (World Precision Instruments). The signals were digitized and recorded with a DigiData 1550A digitizer and AxoScope software (Axon Instruments).

Mestre H., Tithof J., Du T., Song W., Peng W., Sweeney A.M., Olveda G., Thomas J.H., Nedergaard M, & Kelley D.H. (2018). Flow of cerebrospinal fluid is driven by arterial pulsations and is reduced in hypertension. Nature Communications, 9, 4878.

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Electrophysiological Data Acquisition

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Recordings were carried out at room temperature using a MultiClamp 700B amplifier (Axon Instruments), digitized using a Digidata 1550A (Axon Instruments) interface, and visualized in pClamp software (Axon Instruments). Data were filtered at 1 kHz and sampled at 10 kHz.

Buchanan K.L., Rupprecht L.E., Kaelberer M.M., Sahasrabudhe A., Klein M.E., Villalobos J.A., Liu W.W., Yang A., Gelman J., Park S., Anikeeva P, & Bohórquez D.V. (2022). The preference for sugar over sweetener depends on a gut sensor cell. Nature Neuroscience, 25(2), 191-200.

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Intracranial Pressure Measurement Protocol

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Intracranial pressure was measured through a catheter placed in the CM and connected to a pressure transducer and monitor (World Precision Instruments). The signals were digitized and recorded with a Digidata 1550A digitizer and AxoScope software (Axon Instruments).

Hablitz L.M., Vinitsky H.S., Sun Q., Stæger F.F., Sigurdsson B., Mortensen K.N., Lilius T.O, & Nedergaard M. (2019). Increased glymphatic influx is correlated with high EEG delta power and low heart rate in mice under anesthesia. Science Advances, 5(2), eaav5447.

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Ionic Current Measurement and Analysis

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Ionic currents were collected either at a 100 kHz sampling rate with a 10 kHz low pass filter using patch clamp amplifier Axon Axopatch 200B, with digitizer DigiData 1550A from Axon Instruments Inc. PClamp 10.4 software and an in-house developed LabView program were used for recording the ionic current data. The data were analyzed using MATLAB based software OpenNanopore, developed by Laboratory of Nanoscale Biology (LBEN) of École polytechnique fédérale de Lausanne (EPFL). OpenNanopore fits abrupt stepwise signals in the presence of Gaussian noise with a level threshold of 200 pA. Statistic analysis was carried out in OriginPro 2015, in which the Levenberg–Marquardt algorithm was used for Gaussian fitting.

Biswas S., Song W., Borges C., Lindsay S, & Zhang P. (2015). Click Addition of a DNA Thread to the N-termini of Peptides for Their Translocation through Solid-State Nanopores. ACS nano, 9(10), 9652-9664.

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Electrophysiological Data Acquisition

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Recordings were performed at room temperature using a MultiClamp 700B amplifier (Axon Instruments), digitized using a Digidata 1550A (Axon Instruments) interface and visualized in pClamp software (Axon Instruments). Data were filtered at 1 kHz and sampled at 10 kHz.

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Patch-Clamp Analysis of Interstitial Cells

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ICCs were identified as cells with green fluorescent Kit proteins (ACK₂) labeled with Alexa Fluor 488 (Molecular Probes, Eugene, OR, USA). using an inverted fluorescence microscope. The standard whole cell patch clamp configuration was employed to record membrane currents (voltage clamp) and potentials (current clamp). We recorded from small ICC clusters, as spontaneous inward currents from small groups of cells are more robust and regular than from single cells. Currents or potentials were amplified with an Axopatch 200B patch-clamp amplifier (Axon Instruments, Foster, CA, USA) and digitized with a 16-bit analog to digital converter (Digidata 1550A; Axon Instruments, Foster, CA, USA). Data was sampled at 4 kHz and filtered at 5 kHz using an eight-pole Bessel filter for whole cell experiments. All data was analyzed using clampfit (pCLAMP version, 10.2; Axon Instruments) and Graph Pad Prism (version 4.0) software. All experiments were performed at 30℃.

Park I.K., Kim J.H., Park C.G., Kim M.Y., Parajuli S.P., Hong C.S., Choi S, & Jun J.Y. (2018). Effects of ATP on Pacemaker Activity of Interstitial Cells of Cajal from the Mouse Small Intestine. Chonnam Medical Journal, 54(1), 63-71.

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