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Supporting Information:SweepStat: A Build-it-Yourself, Two-Electrode Potentiostatfor Macroelectrode and Ultramicroelectrode StudiesMatthew W. Glasscotta, Matthew D. Verbera, Jackson Halla, Andrew D. Pendergasta, Collin J. McKinneya, and Jeffrey E. Dicka,b*aDepartment of Chemistry, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599-3290, USAbLineberger Comprehensive Cancer Center, School of Medicine, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599-3290, USA* To whom correspondence should be addressed: jedick@email.unc.edu??Table of ContentsPagePedagogical and Technical Two and Three Electrode System Details (Figure S1-S2)S-2SweepStat Enclosure Design (Figure S3)S-5Materials for Electrochemical ExperimentsS-5Pedagogical Overview of Macro and Microelectrochemical Measurements (Figure S4)S-6Finite-Element Simulation and Commercial Potentiostat Comparison (Figure S5)S-6List of SweepStat Components (Table S1)S-9Complete Circuit Diagram (Figure S6)S-10Printed Circuit Board Layout and Completed SweepStat Image (Figure S7)S-11Faraday Cage Construction (Figure S8)S-12Dummy Cell Experiment (Figure S9)S-13Step-by-Step Assembly and Testing ProcedureS-14COMSOL Finite Element Simulation Parameters (Table S2)S-16 Pedagogical and Technical Two and Three Electrode System DetailsWithin the field of analytical chemistry, electrochemistry is uniquely capable of extremely high sensitivity,PEVuZE5vdGU+PENpdGU+PEF1dGhvcj5aaG91PC9BdXRob3I+PFllYXI+MjAxODwvWWVhcj48UmVj
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ADDIN EN.CITE.DATA 13-17 Electrochemical measurements can be used to identify various analyte characteristics, such as diffusion coefficient, analyte concentration, and the number of electrons transferred, following the general reduction of a species O to a species R through the addition of n electrons and assuming reactants and products are freely-diffusing species: O+ne-?R Eq. S1right2450827Figure S1 – Reaction scheme and cyclic voltammogram for ferrocene methanol oxidation to ferrocenium methanol. Starting point was 0 V vs. Ag/AgCl, plotted in classical convention. Working and counter electrodes were a 1.5 mm glassy carbon disk and glassy carbon rod, respectively. Scan rate was 50 mV/s.00Figure S1 – Reaction scheme and cyclic voltammogram for ferrocene methanol oxidation to ferrocenium methanol. Starting point was 0 V vs. Ag/AgCl, plotted in classical convention. Working and counter electrodes were a 1.5 mm glassy carbon disk and glassy carbon rod, respectively. Scan rate was 50 mV/s.The ability to accurately measure the signal generated from such a reaction depends on the electrochemical instrumentation employed. Modern electrochemical measurements are generally conducted using three-electrode potentiostats, which employ a working electrode (WE), a counter electrode (CE), and a reference electrode (RE) to monitor the current or potential produced from an electrochemical reaction. ADDIN EN.CITE <EndNote><Cite><Author>Elgrishi</Author><Year>2018</Year><RecNum>109</RecNum><DisplayText><style face="superscript">18</style></DisplayText><record><rec-number>109</rec-number><foreign-keys><key app="EN" db-id="f2w5pdsdvdww2berxd3vzp96wwdpex9s2swx" timestamp="1543875429">109</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Elgrishi, Noémie</author><author>Rountree, Kelley J.</author><author>McCarthy, Brian D.</author><author>Rountree, Eric S.</author><author>Eisenhart, Thomas T.</author><author>Dempsey, Jillian L.</author></authors></contributors><titles><title>A Practical Beginner’s Guide to Cyclic Voltammetry</title><secondary-title>Journal of Chemical Education</secondary-title></titles><periodical><full-title>Journal of Chemical Education</full-title></periodical><pages>197-206</pages><volume>95</volume><number>2</number><dates><year>2018</year><pub-dates><date>2018/02/13</date></pub-dates></dates><publisher>American Chemical Society</publisher><isbn>0021-9584</isbn><urls><related-urls><url> Of particular interest is a technique known as cyclic voltammetry (CV), where the current (i) is measured while scanning the potential (E) from E1 E2 E1. In this manuscript, we also use CV interchangeably with cyclic voltammogram, which is the output of a cyclic voltammetry experiment, demonstrated in Figure S1. Characteristic peak position and CV shape allow for characterization of both single and multi-analyte solutions. The example CV in Figure S1 shows the oxidation of ferrocene methanol to ferrocenium methanol and subsequent reduction back to ferrocene methanol in an aqueous solution. The interested reader is referred to prior educational literature describing the CV shape.PEVuZE5vdGU+PENpdGU+PEF1dGhvcj5FbGdyaXNoaTwvQXV0aG9yPjxZZWFyPjIwMTg8L1llYXI+
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ADDIN EN.CITE.DATA 18, 19 Before moving forward, it is necessary to comment on the sign convention in electrochemistry. While the IUPAC convention is to plot anodic (positive) potentials to the right, cathodic (negative) potentials to the left, the opposite convention also persists in the literature. This convention comes from the Heyrovsky days of polarography. In polarography, the hanging mercury drop electrode was most frequently used, and this electrode itself can be easily oxidized. As a result, only cathodic (negative potentials, negative current) electrochemistry could be physically observed, requiring data to be plotted in the third quadrant. Therefore, the previously discussed convention was established to conserve space by reversing the signs and plotting cathodic data in the first quadrant. In the case of voltammetry with a three-electrode cell, the potential applied at the WE, which drives the redox reaction of interest, is compared to a known redox chemistry at the RE (e.g., normal hydrogen electrode (NHE), saturated calomel electrode (SCE), or silver/silver chloride (Ag/AgCl) electrode). This comparison establishes a reference point in the voltammogram against which measurements can be reproduced. The CE completes the circuit in this configuration, allowing current to flow from the WE to the CE or vice versa. While this conventional setup is adequate for many experiments, the circuitry required is complex. Furthermore, optimizing the sensitivity of these instruments for applications such as neurological analysis through fast scan cyclic voltammetry (FSCV), ADDIN EN.CITE <EndNote><Cite><Author>Robinson</Author><Year>2003</Year><RecNum>112</RecNum><DisplayText><style face="superscript">20</style></DisplayText><record><rec-number>112</rec-number><foreign-keys><key app="EN" db-id="f2w5pdsdvdww2berxd3vzp96wwdpex9s2swx" timestamp="1543875543">112</key></foreign-keys><ref-type name="Book">6</ref-type><contributors><authors><author>Robinson, Donita</author><author>Venton, B.</author><author>L A V Heien, Michael</author><author>Mark Wightman, R.</author></authors></contributors><titles><title>Detecting subsecond dopamine release with fast-scan cyclic voltammetry in vivo</title><alt-title>Clinical chemistry</alt-title></titles><pages>1763-73</pages><volume>49</volume><dates><year>2003</year></dates><urls></urls></record></Cite></EndNote>20 or single atom measurements through nanoelectrochemistry,PEVuZE5vdGU+PENpdGU+PEF1dGhvcj5EaWNrPC9BdXRob3I+PFllYXI+MjAxNTwvWWVhcj48UmVj
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ADDIN EN.CITE.DATA 24-26 elevates the instrument cost to between $4,000 and $40,000 USD. This high price point hinders the universal incorporation of electrochemistry as an undergraduate or secondary education instructive experience. Thus, reducing the cost of such instrumentation may facilitate experimental opportunities at these levels of education and ensure electrochemistry can reach every student.Historically, the topology for a potentiostat used to perform CVs is similar to that shown in Figure S2a. This circuit is the basis for performing CV experiments using a three-electrode cell. A Randles Cell model is used in this circuit, which only accounts for electrochemical activity at the WE. In this case, Rs represents the sum of the solution resistance, which depends on the conductivity of the solution and the distance between the WE and RE, and the resistance due to electrode size and conductivity, Zelectrode. ADDIN EN.CITE <EndNote><Cite><Author>Newman</Author><Year>1966</Year><RecNum>369</RecNum><DisplayText><style face="superscript">27</style></DisplayText><record><rec-number>369</rec-number><foreign-keys><key app="EN" db-id="p9z0eddfnz9ssre0te5p0r0s2wpzprtae9t9" timestamp="1524615798">369</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Newman, J.</author></authors></contributors><titles><title>RESISTANCE FOR FLOW OF CURRENT TO A DISK</title><secondary-title>Journal of the Electrochemical Society</secondary-title></titles><periodical><full-title>Journal of the Electrochemical Society</full-title><abbr-1>J. Electrochem. Soc.</abbr-1></periodical><pages>501-&</pages><volume>113</volume><number>5</number><dates><year>1966</year></dates><isbn>0013-4651</isbn><accession-num>WOS:A19667648900021</accession-num><urls><related-urls><url><Go to ISI>://WOS:A19667648900021</url></related-urls></urls><electronic-resource-num>10.1149/1.2424003</electronic-resource-num></record></Cite></EndNote>27 It is important to minimize Zelectrode for the CE with respect to the WE by using a CE that is significantly larger than the WE and made of a material which allows free transfer of electrons from the electrode to the ionic solution. The charge transfer resistance resulting from the heterogeneous electron transfer process at the WE is represented as Rct, while Cdl denotes the double-layer capacitance of the electrode. Although the Randles Cell model is likely an oversimplification of experimental reality, it serves a pedagogical purpose in demonstrating which components contribute to resistance and capacitance within an electrochemical cell. This understanding ultimately allows one to design an effective electrochemical cell. Unfortunately, using a RE, as depicted in Figure S2a, has several disadvantages:Feedback in combination with the CE and WE electrode impedances alters the frequency response of the potentiostat. This can result in signal instability at high scan rates.right634274Figure S2 – (a) Electronic schematic of a three-electrode potentiostat system with working electrode (WE), counter electrode (CE), and reference electrode (RE) employed to measure the response of a Randles Cell solution model. (b) Electronic schematic of a two-electrode potentiostat system where the voltage ramp is driven through the op-amp’s virtual connection. 00Figure S2 – (a) Electronic schematic of a three-electrode potentiostat system with working electrode (WE), counter electrode (CE), and reference electrode (RE) employed to measure the response of a Randles Cell solution model. (b) Electronic schematic of a two-electrode potentiostat system where the voltage ramp is driven through the op-amp’s virtual connection. Circuit complexity increases with the use of a reference electrode. The circuit diagrams in Figure S2 are simplified versions. Actual circuitry must include additional elements to compensate for frequency response changes and to prevent oscillation. The curious reader is referred to the first edition of Electrochemical Methods: Fundamentals and Applications by Bard and Faulkner for complete circuitry. ADDIN EN.CITE <EndNote><Cite><Author>Faulkner</Author><Year>2001</Year><RecNum>1134</RecNum><DisplayText><style face="superscript">28</style></DisplayText><record><rec-number>1134</rec-number><foreign-keys><key app="EN" db-id="59wzwvrt1ss2wce9v23xvr0yd9wee2zvzsa2" timestamp="0">1134</key></foreign-keys><ref-type name="Book">6</ref-type><contributors><authors><author>Allen J. Bard and Larry R. Faulkner</author></authors></contributors><titles><title>Electrochemical Methods: Fundamentals and Applications</title></titles><edition>1</edition><dates><year>2001</year></dates><pub-location>New York, New York</pub-location><publisher>John Wiley & Sons</publisher><urls></urls></record></Cite></EndNote>28In Figure S2a of the main text, the potentiostat circuitry consists of a voltage source, Vin, which is applied to amplifier A1. When this circuit is used for CVs, amplifier A1 is used to apply the voltage ramp to the electrochemical cell through the CE. This results in a current (i), which flows through the cell from CE through WE and into the virtual ground of A2, where it is converted to a voltage (Vout) that is proportional to current i. The circuit of Figure S2a is governed by the following equations:ZElectrode=RCT||CDBL=RCT * XCDBLRCT + XCDBL Eq. S2VWE=Vramp-i*(RS+ZElectrode) Eq. S3i=VrampZElectrode+RSEq. S4Vout=-i*REq. S5Equation S2 defines the electrode impedance, ZElectrode, to be the parallel combination of Rct, and Cdl. The application of the voltage, Vin, to the cell causes current to flow from the CE to the WE and into the virtual ground of A2. From equation S3 above, it is apparent that VWE is less than Vin due to the iR voltage drop (often referred to as Ohmic drop) across RS. CV experiments aim to measure the change in i resulting from changes in ZElectrode as a function of VRamp as seen in equation S4. It is important to be able to accurately identify redox potentials by knowing the exact voltage at the WE. In the case where the solution resistance and/or the electrode resistance is large in comparison with the impedance of Rct||Cdl, the iR drop through Rs causes significant error in the potential measured at WE. Compensation for Rs can be accomplished by using a third “reference electrode” in very close physical proximity to the WE. The voltage difference between VRamp and VRef is measured and used to increase VRamp to make up for the iR drop across Rs. This is performed in a feedback operation around A1 which can correct for changes in Rs in real time. Equation S5 describes the transformation of the current into a voltage, Vout. Using a third electrode to compensate for iR drop is important in the following situations:If Rs is large enough in comparison to ZElectrode to cause a significant shift in the redox potential. This manifests in low-conductivity solutions, such as non-aqueous solvents, especially in the absence of supporting electrolyte. If the CE is smaller than the WE causing non-linear iR drops in series with RS that must be eliminated. Using a reference electrode allows for real-time correction of iR drops across RS and an imperfect CE.The primary design goals of the SweepStat included low cost, reduced circuit complexity, and robust operation (i.e., lack of instability such as circuit oscillation). With these goals in mind, and the limitations stated above, a two-electrode system was pursued instead of the more complex three-electrode potentiostat. In order to minimize the effects of Rs on measurements, the SweepStat educational experiments have been designed around classical electrochemical reactions using highly conductive ionic solutions, low impedance WEs, and employing a large CE. These precautions eliminate iR drop in the electrochemical cell and its associated errors. While a general reference electrode such as Ag/AgCl may be used in the UME experiments as a counter/reference electrode, using larger electrodes in these schemes may pass too much current through the reference such that its equilibrium is disturbed, and its reference potential drifts. Therefore, for referenced experiments, a redox standard (i.e. a molecule with a known E0’) may be introduced into the solution to “calibrate” the output. In this scheme, inert conducting materials such as glassy carbon or platinum may be used as a counter electrode to facilitate the passage of large amounts of current. For pedagogical purposes, the concepts of electroactive analyte flux to the electrode surface may still be demonstrated without the use of a reference electrode or redox standard. It is important to realize that without a defined chemical equilibrium against which to reference the reaction, the potential response (i.e. the x-axis of the CV) is relative to the experiment and cannot be related to the electrochemical series. In this case, the term quasi-reference electrode (QR) is used to indicate a reference electrode without a defined equilibrium. We elected to simplify the circuitry further by employing a voltage-follower concept combined with the current to voltage converter, eliminating the need for a feedback Op-Amp (A1) and reducing the entire circuit to a single Op-Amp as shown in Figure S2b. In this case, Vramp is applied to the non-inverting input of the left454Figure S3 – (a) Autodesk Fusion 360 3D rendering of customized SweepStat enclosure box. Total print time <10 hours. (b) Printed enclosure containing SweepStat circuitry highlighting two-electrode interface, where the red and black wires correspond to the working and counter electrodes, respectively. (c) Enclosure top removed to reveal internal circuitry.00Figure S3 – (a) Autodesk Fusion 360 3D rendering of customized SweepStat enclosure box. Total print time <10 hours. (b) Printed enclosure containing SweepStat circuitry highlighting two-electrode interface, where the red and black wires correspond to the working and counter electrodes, respectively. (c) Enclosure top removed to reveal internal circuitry.current to voltage converter (A). As such, the Op-Amp will drive Vout to a value which induces the voltage at the non-inverting input to equal the voltage at the inverting input, thus forcing the WE to exactly follow Vramp (hence “voltage-follower”). When the WE is raised to Vramp, i flows from the WE to the CE which is grounded and Vout follows equation S6:Vout=Vramp-(i*Rf)Eq. S6Therefore, to directly plot the resulting i-V or i-t curve, Vramp must be subtracted from Vout, which is accomplished in the circuitry (Figure S6).SweepStat Enclosure DesignThe SweepStat instrumentation box was designed and fabricated using commercial CAD software and 3D-printers (Ultimaker S3 with CPE filament). A simple enclosure was created in Autodesk Fusion 360 as presented in Figure S4a, however, a similar design could be generated using numerous open-source software packages for 3D modelling (e.g. openSCAD). A completed SweepStat device is shown in Figure S4b, highlighting the electrode leads, on/off switch, and USB port for connection to the LabView interface. A labeled device is also included in the Supporting Information. Within this two-electrode framework, the red and black leads represent the WE and combined QCE/RE, respectively. Figure S4c shows the device with the top half of the enclosure removed to reveal the completed SweepStat PCB. Materials for Electrochemical ExperimentsPotassium hexacyanoferrate (II) trihydrate (98.5%), potassium hexacyanoferrate (III) (>99.0%), and potassium chloride (>99.0%) were purchased from Sigma-Aldrich and used without further purification. Hydroxymethylferrocene (97%) was purchased from Alfa Aesar and used without further purification. All solutions were prepared with nanopure water (18.2 mΩ?cm). Macrodisk experiments were conducted with 1.5 mm radius glassy carbon electrodes and UME experiments were conducted with 5.0 ?m radius Pt electrodes purchased from CH Instruments. In all experiments, the combined reference/counter electrode was a glassy carbon rod. SweepStat electrochemical measurements were verified with a Model 601E CH Instruments Electrochemical Workstation with a Ag/AgCl aqueous RE and a glassy carbon CE. Pedagogical Overview of Macro and Microelectrochemical Measurementsleft12791Figure S4 – Schematic representation comparing the diffusion profiles about macro and ultramicroelectrodes. From this representation, a semi-infinite linear diffusion profile will result in a colloquial duck shape voltammogram, whilst an ultramicroelectrode’s radial diffusion profile will result in a sigmoidal voltammogram.00Figure S4 – Schematic representation comparing the diffusion profiles about macro and ultramicroelectrodes. From this representation, a semi-infinite linear diffusion profile will result in a colloquial duck shape voltammogram, whilst an ultramicroelectrode’s radial diffusion profile will result in a sigmoidal voltammogram.Electrochemistry at the interface of large metal electrodes has been investigated for centuries. ADDIN EN.CITE <EndNote><Cite><Author>Faulkner</Author><Year>2001</Year><RecNum>1134</RecNum><DisplayText><style face="superscript">28</style></DisplayText><record><rec-number>1134</rec-number><foreign-keys><key app="EN" db-id="59wzwvrt1ss2wce9v23xvr0yd9wee2zvzsa2" timestamp="0">1134</key></foreign-keys><ref-type name="Book">6</ref-type><contributors><authors><author>Allen J. Bard and Larry R. Faulkner</author></authors></contributors><titles><title>Electrochemical Methods: Fundamentals and Applications</title></titles><edition>1</edition><dates><year>2001</year></dates><pub-location>New York, New York</pub-location><publisher>John Wiley & Sons</publisher><urls></urls></record></Cite></EndNote>28 When used for CV experiments, these macroelectrodes produce a colloquial duck shape in the resultant plot of current as a function of applied potential, which results from the changing concentration profiles of analyte at the electrode surface. For instance, when a potential sufficient to drive the reduction of a freely diffusing analyte O is applied to the electrode, the surface concentration of O decrease as it is converted to R. At the same time, O is continuously diffusing to the electrode surface from the bulk solution. Due to the large, planar area of the electrode, the diffusion path of O is essentially perpendicular to the electrode surface at any given point. This concept is termed semi-infinite linear diffusion, where semi-infinite represents the idea that the electrode extends infinitely in all directions from a molecular perspective as O diffuses to the surface (except when that molecule approaches the very edge of the electrode, hence semi-infinite), and linear indicates the two-dimensional perpendicular diffusion path of O. The concentration profiles about the electrode surface generated by semi-infinite linear diffusion behavior are changing with time (i.e., the exhaustion layer for O is expanding outward from the electrode), giving rise to the oxidation and reduction peak shape in classical voltammograms. As the electrode is made smaller, the fraction of O diffusing to the edge of the electrode increases. Eventually, a new diffusion profile is generated where O may diffuse to the electrode surface radially, greatly accelerating the rate of replenishment for O at the surface following reduction to R. Thus, the concentration profiles adjacent to the electrode surface do not change with time at commonly used scan rates (i.e. 50-100 mV/s), simplifying the mathematical treatment. This new diffusion profile manifests in the voltammogram as a sigmoidal curve, where the final current magnitude is termed the diffusion-limited current. It should be noted that steady-state behavior is not solely dependent on electrode size but also the scan rate of the experiment. At sufficiently fast scan rates, the steady-state voltammogram for a microelectrode will develop “peaks” characteristic of macro electrodes. This is due to the dependence on diffusion layer thickness on scan rate. At sufficiently small diffusion layers, mass transfer resembles semi-infinite linear diffusion. Similarly, at a macro electrode, scanning sufficiently slow may produce steady-state voltammograms as the potential changes so slowly that the reaction becomes diffusion controlled. These essential electrochemical concepts, further illustrated in Figure S4, are readily demonstrated using the SweepStat’s low and high gain capabilities, providing experimental context to the abstract principle of analyte diffusion.Finite-Element Simulation and Commercial Potentiostat Comparison While the SweepStat offers a facile method to perform classical electrochemistry experiments and extract chemical and physical properties with good agreement to literature values, it is imperative that the observed electrochemical measurements agree with industry standard workstations and simulation packages. The CH Instruments 601e electrochemical workstation offers an excellent commercially available instrumentation package for voltammetry and amperometry using a three-electrode cell at a cost of >$5,000 USD. In Figure S5a, an overlay of macrodisk voltammetry of 75 ?M ferrocenemethanol, as discussed in Section 3.1, is presented corresponding to the SweepStat data (green) and the CHI data (blue), demonstrating similar CVs and validating the measurement capabilities of the SweepStat. Additionally, a 1-dimensional COMSOL 5.3a finite element simulation was conducted with the resultant CV plotted in Figure left363Figure S5 – (a) CV of 75 ?M ferrocenemethanol on a 1.5 mm glassy carbon disk using a SweepStat (green) and CHI E600 Workstation (blue) demonstrating good agreement that can be modelled with COMSOL (red). CHI results are shifted -150 mV to account for reference variation. (b) CV of 7.5 mM potassium ferrocyanide on a 5 μm platinum UME on the two potentiostats and with COMSOL. Sweep rate was 50 mV/s. CHI results are shifted -60 mV to account for reference variation.Figure S5 – (a) CV of 75 ?M ferrocenemethanol on a 1.5 mm glassy carbon disk using a SweepStat (green) and CHI E600 Workstation (blue) demonstrating good agreement that can be modelled with COMSOL (red). CHI results are shifted -150 mV to account for reference variation. (b) CV of 7.5 mM potassium ferrocyanide on a 5 μm platinum UME on the two potentiostats and with COMSOL. Sweep rate was 50 mV/s. CHI results are shifted -60 mV to account for reference variation.S5a (red). The parameters used for these simulations are presented in Table S2. It is important to note that due to the lack of a separate reference electrode in the SweepStat design and thus the use of a glassy carbon QCE/RE, the E1/2 values for the SweepStat and CHI CV scans were separated by 150 mV, and Figure S5a demonstrates a -150 mV correction to the potential at the CHI workstation. 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ADDIN EN.CITE.DATA 29-31SweepStat Instructional MeritOne unique advantage of the open-source SweepStat compared to other low-cost potentiostats or commercially available industrial potentiostats is the ease of integration into common chemistry curricula coupled with its capability of making measurements on UMEs. While the SweepStat has a number of instrumental limitations including two narrow signal gain ranges, Teensy Firmware integration for interfacing with LabView, and a two-electrode cell leading to floating reference potentials, the underlying instructional value of experimental electrochemical measurements is comparable to high-end commercial instrumentation. By combining low cost (<$55 USD), minimal prerequisite technical skills for assembly, an open-source box design, open-source LabView GUI, and ample classical electrochemical experiments, the SweepStat offers a unique all-in-one electrochemical package that could be integrated into undergraduate chemistry or secondary education courses. Importantly, the open-source nature of the SweepStat lends itself well to future advancement stemming from student innovation. For example, while the current SweepStat design is capable of macro- and micro-electrode experimentation, modifications to the circuitry could allow significantly higher sampling frequencies, enabling FSCV measurements. Furthermore, several classical electrochemical methods are not currently included in the SweepStat circuitry including potentiometry and variable waveform voltammetry. Although simultaneous integration of these techniques would ultimately increase the price of fabrication, a coupled chemical engineering and electrochemistry class could reasonably design circuitry, prepare LabView programs, and conduct experiments with data output competitive with commercial potentiostat packages. Importantly, the current low-cost fabrication of the SweepStat is attractive as an instructional demonstrative tool within a lecture classroom. This may be accomplished by providing experimental validation for electrochemical relationships studied in class, guiding students through problem-solving techniques, and calculating a wide range of chemical and physical properties of freely diffusing molecules. Beyond demonstrative instruction, the SweepStat could reasonably be used as a low-cost alternative for electrochemical workstations in low-resource settings, thereby facilitating research opportunities for all interested parties. Table S1 – List of SweepStat components. All parts are generic and may be purchased in any hardware store unless otherwise noted. Board LabelQtyDescriptionPricePCB1Printed Circuit Board, 1/2Express PCB$19.48U11Dual OpAmp,$3.16 TI LMC6484B1, B22Coin Cell Holder,Keystone #103$1.71Batt2Battery, 3V, CR2032$0.98J1, P11Connector, 2 pin Jack and plug w/ pigtails, All Electronics CON-242$0.85S11DPST Switch$1.62U21Arduino Teensy$19.95JP1, JP222 pin header jumper$0.12JP1, JP222 pin header$0.10U21Pin Strip Socket 29 pins total$1.20C1. C4, C530.1uF capacitor$0.42C211pF capacitor$0.14C3130pF capacitor$0.14R1130k resistor$0.10R2, R3, R6, R7, R10, R13610k resistor$0.60R4, R82100M resistor$2.44R5, R921M resistor$0.20R1113.6k resistor$0.10R1211.2k resistor$0.10R14, R15, R18, R194100k resistor$0.40R16, R1721k resistor$0.2041/4" Aluminum Standoffs$0.3544-40 Nuts$0.054#4 Lockwashers$0.054#4 X 0.25" Bolt$0.052Alligator Clips$0.37U1114 Pin DIP socket$0.75 LINK Excel.Sheet.12 "Book1" "Sheet1!R1C1:R30C8" \a \f 4 \h \* MERGEFORMAT Figure S6 - The SweepStat is designed to run on two 3V C2302 button-cell batteries. Voltage ramping and profiling is controlled by a Teensy 3.2 microcontroller chip (E), which is interfaced with a GUI written in LabView, allowing for easy use of the device with minimal training. The Teensy also acts as a digital to analog converter and vice versa. The op amp labeled (A) in Fig. 2 above shifts up the voltage output of the ramp being supplied by the Teensy. (B) subtracts and inverts the ramp. (C) is a current/voltage amplifier which supplies a variable current to the working electrode (F) which is connected at the inverting input. JP1 and JP2 (G) allow for the user to vary the gain of this feedback circuit by adding an additional connection to change the total resistance of the feedback loop. (D) is another level shift inverter which outputs to the Teensy for analog-digital conversion, the result of which is sent to the LabView GUI. Figure S7 - Graphic of the PCB layout of the SweepStat circuit. All parts listed in Table S1 can be soldered onto the PCB board shown to create the functional SweepStat device. The two jumpers controlling the gain setting are circled in red, and the Teensy reset button is circled in blue.Figure S8 – Faraday cage constructed from Styrofoam box and aluminum foil. (a) A Styrofoam box was chosen as the frame for the Faraday cage, though cardboard and other scaffold materials may be used. The external walls of the box, as well as the lid, were wrapped in aluminum foil to act as the grounded conductor. Separate wrapping of the lid allows the box to be easily opened between measurements. (b) Constructed Faraday cage with internal electrochemical cell set up for experimentation. The SweepStat is insulated from the top of the box and connected to the aluminum foil via an alligator clip. This connection facilitates grounding to reduce the effect of electromagnetic radiation. (c) Closed Faraday cage grounded to the SweepStat permits electrochemical measurements free of EM-induced noise.Figure S9 – Graphical interface image showing the result of the dummy cell experiment in which as resistor is placed across the working and counter electrode leads. This resistance gives rise to a linear i-E curve that follows Ohm’s law.Step-by-Step Assembly and Testing ProcedureAssembly InstructionsObtain SweepStat parts as indicated in Table S1.Obtain SweepStat PCB board by downloading Express PCB from the following url: the file SweepStatPCB.pcb attached in the supporting information and place the order for PCBs directly within the program.Use standard soldering techniques to prepare and solder components to the PCB. The PCB has silk-screened reference designators for each component that match the descriptor in the parts list. This information is used to place the components on the board in the correct positions. The project photos can be used to verify correct placement. Sweepstat Software Initial Set-upInstall the Arduino IDE: Teensyduino: the SweepStat Teensy to the computer with a USB cable.Load Teensy firmware (files attached in supporting information):Start the Arudino IDE.Import the LinxPjrcTeensy31 library (Sketch -> Include Library -> Add .ZIP Library…)Import the LinxSerialListener library (Sketch -> Include Library -> Add .ZIP Library…)Open PJRC_Teensy31_Serial_DAC.inoUpload the sketch (-> button in Arduino IDE). The Teensy programmer will run, uploaded the sketch. You can manually reprogram the firmware by pressing the button on the Teensy.Install the LabVIEW Run-Time Engine (2018 SP1, 32-bit): en-us/support/downloads/software-products/download.labview.html#305508Install NI-VISA: en-us/support/downloads/drivers/download.ni-visa.html#305862Install the SweepStat software (executable file attached in supporting information)Running the SweepStat Software1. Turn SweepStat switch to “ON” position and connect Teensy USB to computer configured as described in “SweepStat Software Initial Setup”.2. Open LabView SweepStat program and verify DAC port in LabView GUI corresponds to Teensy microUSB.Troubleshooting: If not connecting to the computer, check the batteries, make sure correct COM port is selected, or reprogram the firmware (step 4, above) on the Teensy board. If the SweepStat is left in the “ON” position for extended periods of time when not in use, the batteries will be drained. Additionally, if a part needs to be replaced, consult the parts list.Dummy Cell Check (Figure S9)1. Clip a 1 MΩ resistor between the leads to represent a dummy cell.2. In the SweepStat LabView program, input -0.3 V into the initial potential, +0.4 V for the final potential, and 0.001 as the increment. The resulting CV should be linear and double back on itself. If the output is not linear, check the troubleshooting section. Troubleshooting: Set the CV range to -1.5 V to +1.5 V. Using a digital multimeter (DMM), a voltage measurement should be taken at the Teensy’s DAC output (Pin 19) to ground (ground side of R17 resistor is a convenient test point). The resulting potential should sweep from ~0V to ~3.3 V. Subsequently, a DMM measurement at pin 14 on the LMC6484 op-amp should result in a potential from ~ -1.5 V to ~ +1.5 V corresponding to the maximum potential window of the SweepStat. Third, a range of ~ -3 V to ~ +3 V should be observed at pin 7 on the op-amp. Finally, pin 1 and pin 8 on the op-amp should output ~ -1.4 V to ~ +1.4 V and ~ +3 V to ~ +0.2 V, respectively. Macroelectrode CV1. Attach counter electrode (e.g., Pt wire, graphite rod) to the black alligator clip.2. Attach a polished glassy carbon macrodisk working electrode to the red alligator clip. 3. Place the working and counter electrode in a solution of 75 ?M ferrocene methanol in 250 mM KCl. Ensure the electrodes are submerged, but not in contact with the bottom of the container.4. Ensure that the initial potential (-0.3V), the switching potential (+0.4V), and the potential increment in the LabView GUI are correct. The scan rate (mV s-1) is calculated as 50000 * increment. As such, 0.001, 0.0005, and 0.00025 are equal to 50, 25, and 12.5 mV s-1, respectively. Note: Depending on the counter electrode used, the potential window required to collect the complete CV may vary. Initial values provided might need to be altered slightly.5. Select “Run” from the LabView GUI options and allow the CV to run unperturbed until the CV is complete.6. Following scan completion, save the .txt files to a known directory and adjust the scan rate by changing the increment value for subsequent trials.7. Following experiment completion, remove the electrodes from solution and disconnect them from the leads.8. The relationship between peak current and scan rate can be used to calculate the diffusion coefficient as governed by the Randles-Sevcik equation: ip=268,600n3/2CAD1/2v1/2.Macroelectrode Amperometry1. Repeat steps 1-3 in the macroelectrode CV SOP.2. Select the “Amperometry” tab in the LabView GUI and ensure that the initial potential (0V), initial potential hold duration (5s), step potential (+0.6V) and step potential hold duration (25s) are correct.3. Select “Run” from the LabView GUI options and allow the current-time trace to run until completed.4. Save the resultant current-time plot as a .txt file.5. By fitting the Cottrell equation (it= nFAD1/2Cπ-1/2t-1/2) to the exponential decay of the current, the diffusion coefficient of ferrocene methanol can be calculated. Ultramicroelectrode CV1. Remove the two grey jumpers (Figure S7, red) on the SweepStat printed circuit board and return the SweepStat to the enclosure box. This will alter the signal gain and allow for currents produced from ultramicroelectrodes to be measured.2. Attach counter electrode (e.g., Pt wire, graphite rod) to the black alligator clip.3. Attach a polished Pt ultramicroelectrode (5 ?m dia.) working electrode to the red alligator clip. 4. Place the working and counter electrode in a solution of 7.5 mM potassium ferrocyanide in 250 mM KCl. Ensure the electrodes are submerged, but not in contact with the bottom of the container.5. Ensure that the initial potential (-0.3V), the switching potential (+0.4V), and the potential increment (0.001) in the LabView GUI are correct.Note: Depending on the counter electrode used, the potential window required to collect the complete CV may vary. Initial values provided might need to be altered slightly.6. Select “Run” from the LabView GUI options and allow the CV to run unperturbed until completion in a grounded Faraday Cage.Note: While CV characterization on a macrodisk electrode often does not require a grounded Faraday Cage for satisfactory signal-to-noise ratios, it is imperative that UME characterization occur in a grounded Faraday Cage as observed currents can be on the order of nA or hundreds of pA.7. Save the resultant current-potential plot to a known directory. The SweepStat should be returned to the “OFF” position to not drain the batteries and the grey jumpers should be replaced.8. The CV of a ultramicroelectrode produces a steady-state current (iss), which allows for the calculation of the diffusion coefficient for potassium ferrocyanide using iss=SOL Finite Element ModellingFinite element models of cyclic voltammetry at macrodisk electrodes and ultramicroelectrodes were prepared using COMSOL Multiphysics 5.4 with modified open-source and in-house models. Cyclic voltammetry at a macroelectrode was simulated using a model available for download from the COMSOL support website (application ID: 12849, Cyclic Voltammetry). In short, a 1-D model of diffusion perpendicular to a point at the electrode surface was used to calculate the concentration profiles of the redox species R and O given: O+ne-?RWith a local current density calculated from the Butler-Volmer equation where: jlocal=nFk0CRe1-αFηRT-COe-αFηRTWhere n is the number of electrons transferred (note: Butler-Volmer kinetics in COMSOL are only valid for systems in which n = 1), F is the Faraday constant, k0 is the heterogeneous rate constant, η is the overpotential relative potential (E) to the formal potential (Ef,) as η = E - Ef, CR is the surface concentration of species R, α is the cathodic transfer coefficient, CO is the surface concentration of species O, R is the universal gas constant, and T is temperature. As a macroelectrode is modelled to follow semi-infinite linear diffusion, the total current can be calculated by multiplying the local current density jlocal by the electrode area A. The geometry was chosen to be sufficiently long to model semi-infinite linear diffusion at:xmax=6DtscanWhere xmax defines the geometric domain, D is the diffusion coefficient, tscan is the time required for a scan through the potential range, and (Dtscan)1/2 is the distance travelled over the scan, generating a domain large enough to model the bulk concentration of analyte. Cyclic voltammetry at an ultramicroelectrode was simulated using an in-house model under 2D-axis symmetric conditions under similar Butler-Volmer conditions. Importantly, at an UME, radial diffusion from the bulk solution is observed with a steady-state concentration profile. As species O is reduced to R, O is nearly-instantaneously replenished as R diffuses to the bulk solution. A cyclic voltammetry boundary condition at the electrode surface with electroanalytical Butler-Volmer local current density (vide supra) was employed with a controlled potential range and scan rate, generating sigmoid shaped cyclic voltammograms. The double layer capacitance at the UME surface can be ignored due to the small electroactive surface area. In all simulations, a transfer coefficient of α = 0.5 was employed. Table S2 contains the simulation parameters employed to generate the fits presented in the main text. In both the macroelectrode and UME simulations, the COMSOL 5.4 Electrochemistry module with the Electroanalysis module with support for integrated species transport and heterogenous electron transfer. COMSOL 5.4 models are available upon request. 1657352540Table S2 – COMSOL Multiphysics parameters used for CV modeling00Table S2 – COMSOL Multiphysics parameters used for CV modelingMacroelectrodeMicroelectrodeParameterValueParameterValueScan Rate25 mV/sScan Rate10 mV/sBulk Concentration75 μMBulk Concentration7.5 mMDiffusion Coefficient 2.00E-11 m2/sDiffusion Coefficient 6.00E-10 m2/sElectrode Radius1.5 mmElectrode Radius6.25 μmDouble Layer Capacitance 1.82 μF/cm2CV Range -0.6 to 0.8 VCV Range -0.6 to 0.8 VMesh qualityFinerMesh qualityNormalFormal Potential0.01 VFormal Potential0.01 VHeterogenous Rate Constant1 m/sHeterogenous Rate Constant0.1 cm/sNumber of Electrons Transferred1Number of Electrons Transferred1References: ADDIN EN.REFLIST 1.Zhou, M.; Dick, J. E.; Hu, K. K.; Mirkin, M. V.; Bard, A. J., Ultrasensitive Electroanalysis: Femtomolar Determination of Lead, Cobalt, and Nickel. Anal. Chem. 2018, 90 (2), 1142-1146.2.Percival, S. J.; Dick, J. E.; Bard, A. J., Cathodically Dissolved Platinum Resulting from the O-2 and H2O2 Reduction Reactions on Platinum Ultramicroelectrodes. Anal. Chem. 2017, 89 (5), 3087-3092.3.Zhou, M.; Dick, J. E.; Bard, A. J., Electrodeposition of Isolated Platinum Atoms and Clusters on Bismuth-Characterization and Electrocatalysis. J. Am. Chem. Soc. 2017, 139 (48), 17677-17682.4.Kim, J.; Dick, J. E.; Bard, A. J., Advanced Electrochemistry of Individual Metal Clusters Electrodeposited Atom by Atom to Nanometer by Nanometer. Accounts of Chemical Research 2016, 49 (11), 2587-2595.5.Dick, J. E.; Lebegue, E.; Strawsine, L. M.; Bard, A. J., Millisecond Coulometry via Zeptoliter Droplet Collisions on an Ultramicroelectrode. Electroanalysis 2016, 28 (10), 2320-2326.6.Dick, J. E.; Bard, A. J., Toward the Digital Electrochemical Recognition of Cobalt, Iridium, Nickel, and Iron Ion Collisions by Catalytic Amplification. J. Am. Chem. Soc. 2016, 138 (27), 8446-8452.7.Dick, J. E., Electrochemical detection of single cancer and healthy cell collisions on a microelectrode. Chem. Commun. 2016, 52 (72), 10906-10909.8.Lebegue, E.; Anderson, C. M.; Dick, J. E.; Webb, L. J.; Bard, A. J., Electrochemical Detection of Single Phospholipid Vesicle Collisions at a Pt Ultramicroelectrode. Langmuir 2015, 31 (42), 11734-11739.9.Dick, J. E.; Renault, C.; Bard, A. J., Observation of Single-Protein and DNA Macromolecule Collisions on Ultramicroelectrodes. J. Am. Chem. Soc. 2015, 137 (26), 8376-8379.10.Dick, J. E.; Bard, A. J., Recognizing Single Collisions of PtCl62- at Femtomolar Concentrations on Ultramicroelectrodes by Nucleating Electrocatalytic Clusters. J. Am. Chem. Soc. 2015, 137 (43), 13752-13755.11.Dick, J. E.; Renault, C.; Kim, B. K.; Bard, A. J., Electrogenerated Chemiluminescence of Common Organic Luminophores in Water Using an Emulsion System. J. Am. Chem. Soc. 2014, 136 (39), 13546-13549.12.Dick, J. E.; Renault, C.; Kim, B. K.; Bard, A. J., Simultaneous Detection of Single Attoliter Droplet Collisions by Electrochemical and Electrogenerated Chemiluminescent Responses. Angew. Chem. Int. Ed. 2014, 53 (44), 11859-11862.13.Glasscott, M. W.; Pendergast, A. D.; Dick, J. E., A Universal Platform for the Electrodeposition of Ligand-Free Metal Nanoparticles from a Water-in-Oil Emulsion System. ACS Applied Nano Materials 2018.14.Pendergast, A. D.; Glasscott, M. W.; Renault, C.; Dick, J. E., One-step electrodeposition of ligand-free PdPt alloy nanoparticles from water droplets: Controlling size, coverage, and elemental stoichiometry. Electrochem. Commun. 2019, 98, 1-5.15.Bard, A. J. F., Larry R. , Electrochemical Methods: Fundamentals and Applications, 2nd EditionWiley: 2000.16.Xiao, X.; Bard, A. J., Observing Single Nanoparticle Collisions at an Ultramicroelectrode by Electrocatalytic Amplification. Journal of the American Chemical Society 2007, 129 (31), 9610-9612.17.Bard, A. J.; Zhou, H.; Kwon, S. J., Electrochemistry of Single Nanoparticles via Electrocatalytic Amplification. Israel Journal of Chemistry 2010, 50 (3), 267-276.18.Elgrishi, N.; Rountree, K. J.; McCarthy, B. D.; Rountree, E. S.; Eisenhart, T. T.; Dempsey, J. L., A Practical Beginner’s Guide to Cyclic Voltammetry. Journal of Chemical Education 2018, 95 (2), 197-206.19.Kissinger, P. T.; Heineman, W. R., Cyclic voltammetry. J. Chem. Educ. 1983, 60 (9), 702.20.Robinson, D.; Venton, B.; L A V Heien, M.; Mark Wightman, R., Detecting subsecond dopamine release with fast-scan cyclic voltammetry in vivo. 2003; Vol. 49, p 1763-73.21.Cannes, C.; Kanoufi, F.; Bard, A. J., Cyclic voltammetry and scanning electrochemical microscopy of ferrocenemethanol at monolayer and bilayer-modified gold electrodes. Journal of Electroanalytical Chemistry 2003, 547 (1), 83-91.22.Bard, A. J.; Fan, F.-R. F.; Pierce, D. T.; Unwin, P. R.; Wipf, D. O.; Zhou, F., Chemical Imaging of Surfaces with the Scanning Electrochemical Microscope. Science 1991, 254 (5028), 68.23.Barker, A. L.; Macpherson, J. V.; Slevin, C. J.; Unwin, P. R., Scanning Electrochemical Microscopy (SECM) as a Probe of Transfer Processes in Two-Phase Systems:? Theory and Experimental Applications of SECM-Induced Transfer with Arbitrary Partition Coefficients, Diffusion Coefficients, and Interfacial Kinetics. The Journal of Physical Chemistry B 1998, 102 (9), 1586-1598.24.Dick, J. E.; Bard, A. J., Recognizing Single Collisions of PtCl6 2? at Femtomolar Concentrations on Ultramicroelectrodes by Nucleating Electrocatalytic Clusters. Journal of the American Chemical Society 2015, 137, 13752-13755.25.Zhou, M.; E. Dick, J.; J. Bard, A., Electrodeposition of Isolated Platinum Atoms and Clusters on Bismuth – Characterization and Electrocatalysis. 2017; Vol. 139.26.Glasscott, M. W.; Dick, J. E., Direct Electrochemical Observation of Single Platinum Cluster Electrocatalysis on Ultramicroelectrodes. Anal. Chem. 2018, 90 (13), 7804-7808.27.Newman, J., RESISTANCE FOR FLOW OF CURRENT TO A DISK. J. Electrochem. Soc. 1966, 113 (5), 501-&.28.Faulkner, A. J. B. a. L. R., Electrochemical Methods: Fundamentals and Applications. 1 ed.; John Wiley & Sons: New York, New York, 2001.29.Glasscott, M. W.; Dick, J. E., Fine-Tuning Porosity and Time-Resolved Observation of Nucleation and Growth of Single Platinum Nanoparticles. ACS Nano 2019.30.Glasscott, M. W.; Pendergast, A. D.; Choudhury, M. H.; Dick, J. E., Advanced Characterization Techniques for Evaluating Porosity, Nanopore Tortuosity, and Electrical Connectivity at the Single-Nanoparticle Level. ACS Appl. Nano Mater. 2019, 2 (2), 819-830.31.Glasscott, M. W.; Pendergast, A. D.; Goines, S.; Bishop, A. R.; Hoang, A. T.; Renault, C.; Dick, J. E., Electrosynthesis of high-entropy metallic glass nanoparticles for designer, multi-functional electrocatalysis. Nat. Commun. 2019, 10 (1), 2650. ................
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