Team:UC Davis/Electrochemistry Electrode Choice

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UC Davis iGEM 2014

Electrode Choice

Electrode Choice

System Optimization

System Optimization

Enzyme Tests

Enzyme Tests

Which electrode to use?

Selecting the electrode was the first step in building our electrochemical system. In the selection process, many features were considered including price, portability, accuracy, solution volume, and compatibility with our hardware and software signal processing device. Our initial idea was to build our own electrode. However, after reviewing many literatures, we have concluded that purchasing one would be the smarter choice for our short time constraints and enhancing accuracy of chemical detection. We narrowed down our choice to one of the commercial electrodes which already has been proved to detect small concentration of NADH. One was DropSens, and the other was Pine Instrument. These two electrode have features which may or may not be suitable for our system.

Electrode Schema and Diagram

DropSens Vs. Pine Instrument:

DropSens Pine Instrument
Pros:
  1. Size and Portability:
    1. Electrode has dimensions of only 3.4 x 1.0 x 0.05cm
    2. Working and counter electrodes are 4mm diameter, made in carbon surface
  2. Price:
    1. Requires 50-60µL volume of solution
    2. Inexpensive than build our own
  3. Disposability
    1. Easily disposable after run experiment
  4. Accuracy
    1. proved to detect NADH in a smaller concentration (see figure(1) - excerpt from DropSens website)


Cons:
  1. Less flexibility:
    1. electrode comes with pre-polymerized surface

Pros:
  1. Flexibility:
    1. gives up an option to self-polymerize electrode with chemicals of our choice
    2. add more adjustment for working/counter/reference electrode which may affect the sensitivity of detection
  2. Re-usable solution cap
    1. closed solution cap prevents contamination of solution
    2. helps portability with its closed-solution cap


Cons:
  1. Price:
    1. Consumes 200 times more volume of solution than Dropsens (10mL vs. 50uL)
    2. closed-solution cap was pricy
  2. Complex electrode pre-treatment
    1. necessity of separate working electrode and counter electrode by gluing surface and wait until it dries up
    2. need to polymerize electrode each time of experiment
  3. Accuracy:
    1. due to complex steps for pre-treatment electrode, more possibility of errors could be caused(i.e: non-complete separation of working and counter electrode results in different signals)

Importance of Mediator & Polymerization

The electrode system consists of a screen-printed chip embedded with three electrodes: the counter, working, and reference electrodes. Though the potential of the reference electrode is kept constant, a voltage bias is applied across the working and counter electrodes to facilitates a buildup of excess positive charge on the working electrode. This buildup induces the directional diffusion of NADH toward the working electrode. The working electrode consists of a carbon ink infused with polymer. Since some polymers have selective oxidative affinity for NADH, we decided to test two polymers that would work the best for our electrode.
Azure A vs. Meldola's Blue comparison Table goes here
Comparison experiment result:

    Polymerization with both compounds, Azure A and Meldola’s Blue both gives some selective oxidative affinity for NADH by lowering the potential required to oxidize NADH at the electrode; therefore, expanding the voltage range favorable for us to conduct experiments at. Both AA and MB has a selective oxidative affinity for NADH, reducing the over potential necessary for NADH oxidation at the working electrode. Although both are advantageous in terms of selectivity for detecting NADH, we have decided to use a polycyclic aromatic monomer dye, Meldola’s Blue (MB) for the following reason:
  1. MB facilitates greater electron deposition and flow.
  2. Azure A demonstrated second order responses, which can be possible indicative of non-trivial chemical interactions at the electrode surface.
  3. MB gave us more reliable readings, especially showed no second order responses.
  4. We can specifically ordered MB-infused electrodes from Dropsens.