The final prototype assay, optimized for serum and represented in a simplified schematic (Fig. 1 A,B) is as follows: (1) 10 µL of serum was deposited on the modified WE and incubated for 45 min to bind CFP10 antigen to the CAb; (2) a PB washing step (repeated three times) was used to remove sample matrix and stop the immunoreaction; (3) 10 µL of commercial-grade, horseradish peroxidase (HRP) labeled anti-CFP10 detector antibody (HRP-DAb) solution was then placed on the WE and incubated for 45 min to sandwich the CFP10 between the CAb and DAb on the WE; (4) a 0.05% SDS washing step (repeated three times), and another PB washing step (repeated three times) was used to remove unbound HRP-DAb; (5) the BETA sensor was then inserted into an “EmStat3 Blue” potentiometer, attached to a laptop running “PSTrace” software (Palmsens, The Netherlands); (6) a 20 µL drop of 3,3′,5,5′-tetramethylbenzidine (TMB)/H2O2 was placed over both electrodes, and a cathodic current of − 0.1 V was applied and the amperometric signal was recorded for 2.5 min29 (link). The current measured from the potentiometer corresponded to the reduction of the oxidized TMB, generated by the coupled reduction of H2O2 catalyzed by the HRP tag on the DAb, which was directly proportional to the amount of the HRP-tagged DAb bound to the sensor, corresponding to the concentration of CFP10 in the sample.
Emstat3 blue
Manufactured by PalmSens
Sourced in Netherlands
The EmStat3 Blue is a compact, portable potentiostat/galvanostat designed for electrochemical measurements. It provides accurate and reliable data acquisition for a wide range of electrochemical techniques.
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8 protocols using emstat3 blue
1
Electrochemical Assay for CFP10 Detection
2
Morphology and Electrochemical Characterization
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Morphology was analyzed with a JEOL 300-S (Tokyo, Japan) scanning electron microscope (SEM), and Raman spectroscopy was analyzed using a microspectrometer (Horiba, LabRAM-HR) (Horiba, Kyoto, Japan). A three-electrode arrangement, including a GCE (ø = 3 mm), a Pt wire, and an Ag/AgCl, were used as the working electrode, counter electrode, and reference electrode, respectively. Cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), and differential pulse voltammetry (DPV) techniques were performed using an EmStat3 + blue (PalmSens, Houten, The Netherlands) potentiostat.
3
Electrochemical Characterization of Glucose Sensors
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Single electrodes were evaluated in a batch cell with a platinum wire used as the counter electrode and Ag/AgCl/saturated KCl as the reference electrode. Glucose was added to the measurement solution before potential application. BFCs were evaluated in an incubator at 36 °C and 70% humidity by dropping 200 μL of 100 mM glucose solution onto the electrodes. All electrodes were used as fabricated. Cyclic voltammetry was performed at a scan rate of 10 mV s−1. Chronoamperometry was performed by applying a potential of 0.2 V. Linear sweep voltammetry was performed at a scan rate of 1 mV s−1. Emstat3 Blue (Palm Sens, Houten, Netherlands) was used as the potentiostat.
4
Potentiometric Hydrogen Sensing at 500°C
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Potentiometric measurements were performed in a stainless-steel reactor. A schematic representation of the experimental setup is shown in Figure 3 .
Sensors were placed in a stainless-steel reactor using feedthroughs to prevent gas leakages (seeFigure 3 ). The temperature was controlled using a clamp-type electrical resistance (1500 W) and a thermocouple (type K) connected to a PID temperature controller (Fuji PXR4). The thermocouple was located as close as possible to the outer side of both electrolytes, and it was assumed as the temperature of the WE for both sensors. The reactor was isolated using glass wool (Kaowool® Blanket, Morgan thermal ceramics, Windsor, UK) to prevent temperature fluctuations.
The hydrogen concentration in the reactor (working electrodes) ranged from 0.02 to 0.5 mbar H2 in Ar by mixing different flow rates of high-purity argon (99.9992%) and a hydrogen calibration mixture (0.1% H2 in Ar), both supplied by Carburos Metálicos (Cornellà de Llobregat—Spain). A calibration mixture of 0.1% H2 in Ar (1 mbar) was used as reference electrodes. Flow rates were controlled using gas mass flow controllers (Bronkhorst EL-FLOW, Ruurlo—Netherlands). Finally, the electrodes were connected to a high-impedance voltmeter (PalmSens EmStat3+ Blue) using platinum wires to measure the potential difference (ΔE) at 500 °C.
Sensors were placed in a stainless-steel reactor using feedthroughs to prevent gas leakages (see
The hydrogen concentration in the reactor (working electrodes) ranged from 0.02 to 0.5 mbar H2 in Ar by mixing different flow rates of high-purity argon (99.9992%) and a hydrogen calibration mixture (0.1% H2 in Ar), both supplied by Carburos Metálicos (Cornellà de Llobregat—Spain). A calibration mixture of 0.1% H2 in Ar (1 mbar) was used as reference electrodes. Flow rates were controlled using gas mass flow controllers (Bronkhorst EL-FLOW, Ruurlo—Netherlands). Finally, the electrodes were connected to a high-impedance voltmeter (PalmSens EmStat3+ Blue) using platinum wires to measure the potential difference (ΔE) at 500 °C.
5
Portable Electrochemical Fentanyl Sensing
6
Cyclic Voltammetry of Redox-Active Compounds
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Cyclic voltammetry was performed with a PalmSens EmStat3 Blue. A 20-ml glass vial was used as an electrochemical cell with a glassy carbon working electrode, platinum mesh counter electrode, and a Ag/AgCl reference electrode (fig. S10A). Voltammograms were recorded from 5 ml of LB medium alone or supplemented with 1 mM methyl viologen, 100 μM pyocyanin, or 1 mM DHNA. The cell was heated to 37°C using a hot plate. Three scans were performed for each sample between using a scan rate of 10 mV/s. Scans were recorded both with and without purging. Purging was performed for 30 min with N2 gas bubbled through a long needle, with said needle being placed in the headspace of the vial when the scan was recorded to maintain anoxic conditions. The third scan for each experiment was recorded, with the corresponding scan of LB medium over the same potential being subtracted from it. Em values were calculated from the purged condition from the potential lying equidistant between the oxidation and reduction peaks. Potentials were converted from millivolts versus Ag/AgCl to millivolts versus SHE by addition of 200 mV.
7
Electrochemical Characterization of Samples
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All electrochemical characterization and data acquisition was performed with electrochemical EmStat3 Blue potentiostat (Palmsens, The Netherlands) controlled by PSTrace software provided by PalmSens. A Barnstead Thermolyne (Type 16,700 Mixer) vortex for homogenization of the solutions, pH/ISE meter (Orion Star A214 model) and IEC CL31R Multispeed centrifuge from Thermo-Scientific were used.
8
Amperometric Measurements with AuNPs-SPCEs
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Amperometric measurements were carried out using an EmStat3 Blue potentiostat (PalmSens, The Netherlands). Experiments were designed and data were controlled by PSTrace version 4.4. All measurements were carried out at room temperature (RT). The transducers employed were gold nanoparticle-modified screen-printed carbon electrodes (AuNPs-SPCEs) (DRP-110GNP, DropSens), formed by a 4-mm-diameter carbon working electrode and printed carbon and silver wires as counter and pseudo-reference electrodes, respectively. A specific cable connector (DRP-CAC, DropSens) interfaced between the AuNPs/SPCEs and the EmStat3 potentiostat. A Raypa steam sterilizer, biological fumehood (Telstar Biostar) and a temperature freezer (New Brunswick Scientific) were also employed.
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