The electrochemical characterization of the lactate biosensor was performed and recorded using a PalmSens4 potentiostat (Supplementary Fig. 2 ) and PSTrace software (PalmSens), respectively. In all experiments using the electrochemical sensor, CA was used as the sensing method, applying a potential step of −0.2 V vs Ag (RE).
Pstrace software
Manufactured by PalmSens
Sourced in Netherlands
PSTrace software is a data acquisition and analysis tool developed by PalmSens for electrochemical measurements. It provides a user-friendly interface for controlling PalmSens electrochemical workstations and managing the collected data.
Automatically generated - may contain errors
9 protocols using pstrace software
1
Lactate Biosensor Electrochemical Characterization
2
Electrochemical Sensor Data Analysis
3
Electrochemical Analysis of Prebiotic Clusters
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Experiments were conducted under ambient conditions, using an EmStat (PalmSens), single-channel potentiostat. Cluster solution mixtures were prepared inside the anaerobic chamber. All solutions were purged with N2 for 15 min prior to analysis, to ensure that no O2 was trapped during the transfer from the anaerobic chamber to potentiostat. A conventional three-electrode cell composed of a glassy carbon working electrode, a Ag/AgCl reference electrode (Alvatek, RE-5B 3 M NaCl, +209 mV vs. standard hydrogen electrode (SHE)) and a platinum wire counter electrode was used for the electrochemical measurements. We did not explicitly add a supporting electrolyte, to minimize interference with cluster formation (as occurred with bicarbonate). However, ~0.01 M NaCl was present in samples through acid-base titrations. Relatively low ionic strength electrolytes, notably NaCl, have been shown to diminish coagulation of FeS nanoparticles91 (link), and NaCl is clearly of prebiotic relevance in marine environments. The working electrode was cleaned with 0.05 μm aluminium oxide slurry on a polishing cloth and thoroughly rinsed with double-distilled H2O before each experiment. Data were recorded using PSTrace software (v5, PalmSens).
4
Amperometric Sensing of Gaseous Analytes
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The conductivity values from the sensor array were determined by amperometric measurements. For this purpose, the current was measured with the PSTrace software (PalmSens BV) at constant voltage (0.1 V) between the electrodes. In this condition, the conductance (G = current/voltage) is directly proportional to current. To correct for differences between device resistances, the conductance is normalized such that ΔG/G0 = (G0 − G)/G0, where G0 is the conductance before exposure to NH3 or TMA and G is the conductance achieved during exposure. In our work, the conductance decreased with analyte exposure and ΔG/G0 was positive. We report the responses as the arithmetic mean of the three replicated sensors for each material.
5
Amperometric Conductivity Sensing
6
Cleaning and Electrochemical Activation of Multichannel PCB Electrodes
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SEP1 BIOTIP multichannel electrode PCB platform (biotip ltd, Bath, UK) were cleaned according to the supplied protocol. This consisted of a 15-minute submersion in a solution of 50 mM KOH in H2O2 30 % (v/v) at room temperature. The PCB was then rinsed with DI water and dried using compressed air. The PCB was then electrochemically cleaned by submerging in 50 mM KOH (DI water as solvent) with an external platinum counter electrode (Metrohm, Runcorn, UK) and 3M NaCl Ag/AgCl reference electrode (IJ Cambria, Llanelli, UK). Cyclic voltammetry was performed on all working electrodes on the PCB using the following parameters: potential window was -1.2 to 0.6 V, scan rate of 0.1 V/s and 15 scans per electrode. The PCB was then rinsed with DI water and dried again using compressed air. All electrochemical measurements were performed using a PalmSens4 potentiostat and the accompanying PSTrace software, both supplied by Palmsens BV (Houten, Netherlands).
7
Quantifying BBB Integrity via TEER
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Transendothelial Electrical Resistance (TEER) of hCMEC/D3 cell monolayers was measured in real time to quantify BBB integrity. In short, 8250 cells in 250 µl growth medium were grown to confluency on collagen-coated 16-well RTCA E-Plates (Agilent, Santa Clara, CA, USA), containing interdigitated gold microelectrodes. The monolayers of hCMEC/D3 cells at 100% confluency were treated for a period of 48 h with TNF-α/IFN-γ solution (10 ng/mL, positive control), and increasing dose (from 5 × 107 to 6 × 108 particles/ml) of uEV, and tEV size-based EV subpopulations. Resistance values (in Ω) were collected at multiple frequencies, ranging from 1 Hz to 1000 kHz at five frequencies per decade, by a palmSens 4 impedance analyzer, controlled by PSTrace software (palmSens BV, Houten, The Netherlands). TEER values were analyzed at a frequency of 6309.57 Hz, reflecting intercellular junctions, and data are depicted as Ω x cm², based on the well’s surface area (0.196 cm2). Data were normalized to the time point 0 h of each condition individually and resistance curves were generated using GraphPad Prism software.
8
Electrochemical Sensing with Screen-Printed Electrodes
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Experiments were carried out with a computer controlled PalmSens potentiostat. All electrochemical data were collected and analysed by the PalmSens PSTrace software. The electrochemical cell was purchased from Dropsens and consisted of a plexiglass flow cell and a disposable screen ink-printed platinum electrode (SPPtE, DRP-150, Dropsens, Italy). They consisted in a platinum disk-shaped (12.6 mm2) working electrode, a paste of silver/silver chloride pseudo-reference and a platinum strip counter electrode, on a ceramic substrate (3.3 cm × 1.0 cm). All the electrochemical measurements were referred to the screen-printed silver pseudo-reference electrode. The screen printed electrodes were interfaced to the potentiostat by a cable connector for SPEs (DRP-CAC, Dropsens, Italy). A Gilson MiniPuls 3 peristaltic pump and a Rheodyne low pressure injector with a 200-µL sample loop completed the apparatus.
9
Electrochemical Characterization of Thin-Film Graphene
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A
platinum counter (Metrohm, Runcorn, UK) and 3 M NaCl Ag/AgCl reference
(IJ Cambria, Llanelli, UK) electrodes were used to perform all measurements
with TFGEs in 5 mM [Fe(CN)6]3–/4– in PBS. Cyclic voltammetry (CV) measurements were obtained by sweeping
a potential range from −0.4 V to +0.6 V at 0.1 V s–1. Differential pulse voltammetry (DPV) measurements mirrored the
CV potential window and scan rate, with a respective potential pulse
and time of 0.025 V and 0.05 s. EIS was measured after 10 min of stabilization
in [Fe(CN)6]3–/4– against an open-circuit
potential from 100 kHz to 0.1 Hz. Eac set
at 0.01 Vrms and Edc at 0 V. 67 frequencies were recorded for characterization
and 50 frequencies for all other experiments.
A PalmSens4 potentiostat
and PSTrace software from Palmsens BV (Houten, the Netherlands) were
used to perform all electrochemical measurements. Nyquist plots were
then fitted to a modified Randles’ equivalent circuit. Subsequent
parameters of interest were extracted and analyzed using Origin and
Matlab.
platinum counter (Metrohm, Runcorn, UK) and 3 M NaCl Ag/AgCl reference
(IJ Cambria, Llanelli, UK) electrodes were used to perform all measurements
with TFGEs in 5 mM [Fe(CN)6]3–/4– in PBS. Cyclic voltammetry (CV) measurements were obtained by sweeping
a potential range from −0.4 V to +0.6 V at 0.1 V s–1. Differential pulse voltammetry (DPV) measurements mirrored the
CV potential window and scan rate, with a respective potential pulse
and time of 0.025 V and 0.05 s. EIS was measured after 10 min of stabilization
in [Fe(CN)6]3–/4– against an open-circuit
potential from 100 kHz to 0.1 Hz. Eac set
at 0.01 Vrms and Edc at 0 V. 67 frequencies were recorded for characterization
and 50 frequencies for all other experiments.
A PalmSens4 potentiostat
and PSTrace software from Palmsens BV (Houten, the Netherlands) were
used to perform all electrochemical measurements. Nyquist plots were
then fitted to a modified Randles’ equivalent circuit. Subsequent
parameters of interest were extracted and analyzed using Origin and
Matlab.
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