Title: In vivo voltammetry for catecholamine analysis
1In vivo voltammetry for catecholamine
analysis
2Outline of this talk
- Why in vivo voltammetry
- What is in vivo voltammetry
- Hows this technique been developed
- What can be achieved in future
3Why in vivo voltammetry
- Subjective reason
- People want to know more about brain
- by studying neurotransmitters.
- Objective realities
- a. neurotransmitters can be studied.
- b. in vivo voltammetry can study brain.
4Study in Brain chemistry
Jonathan A. Stamford, etal Analytical Chemistry
1996, (68) 359A-366A
5Structures of typical catecholamine compounds
dopamine epinephrine, norepinephrine
6Why in vivo voltammetry can study brain?
- 1. microelectrodes are small enough to access the
brain. - 2.carbon fiber electrodes are
- robust against surface degradation.
- 3.fast cyclic voltammetry can provide fast
measurement in ms scale.
7What is in vivo voltammetry
- Basics in cyclic voltammetry(CV)
- Voltammetric analysis of cacholamines in brain.
8Basics in voltammetry
electron flow
How electrodes in electrochemical cell works
At the negative electrode, the cathode,
electrons are given off, and reduction takes
place. At the positive electrode, the
anode, the excess electrons are collected, and
oxidation occurs. This giving-and-taking of
electrons creates an electric current.
anode
cathode
http//nanonet.rice.edu/research/CVtutorial2/
9- How cyclic potential sweep applied
in CV?
The system starts off with an initial potential
at which no redox can take place. At a critical
potential during the forward scan, the
electroactive species will begin to be
oxidized. After reversal of potential scan
direction and depletion of the oxidized species,
the reverse reduction reaction takes place.
http//www-biol.paisley.ac.uk/marco/Enzyme_Electro
de/Chapter1/Ferrocene_animated_CV1.htm
10 Cyclic voltammogram
In CV, the current in the cell is measured
as a function of potential. The potential of an
electrode in solution is linearly cycled from a
starting potential to the final potential and
back to the starting potential. This process, in
turn, cycles the redox reaction. Multiple cycles
can take place. A plot of potential versus
current is then produced.
http//www-biol.paisley.ac.uk/marco/Enzyme_Electro
de/Chapter1/Ferrocene_animated_CV1.htm
11- CV can identify and quantify analytes
Kirk T. Kawagoe, etal Journal of Neuroscience
Methods, 48 (1993) 225-240
12Other voltammetry techniques
Robert D. ONeill, etal Analyst,
May 1994, Vol. 119. 767.
13 Catecholamine analysis by in vivo CV
- How this experiment be carried out in brain?
- Special considerations for in vivo voltammetry
compared with CV in beaker system.
14Experimental set up in vivo voltammetry
Jonathan A. Stamford, etal TINS, Vol. 12,
No. 10, 1989.
15Considerations to In Vivo CV
- In vivo working environment
- ECF (extracellular fluid)
- Requirements for electrodes
- a. minimum tissue damage
- b. stability
- Interferences in measurement
- a. ascorbic acid (AA vitamin C)
- b. DOPAC
16The big concern of in vivo voltammetry
- The Selectivity of catecholamine over
interfering compounds MUST be guaranteed!
Kirk T. etal Journal of
Neuroscience Methods, 48 (1993) 225-240.
17 Hows this technique been developed
- Improvements made based on
- High selectivity
- Good Sensitivity
- Less disturbance imposed on brain function
- Stability of electrodesless poisoning
- Three electrodes are developed carbon paste
electrodes carbon epoxy electrodes carbon fibre
electrodes.
18How high selectivity be achieved?
- Optimization of potential waveform
Crespi, F. etal. (1984) Brain Res. 322,
135-138.
19How high selectivity be achieved?
- Modification on electrodes
- a. coating the electrode surface with selectively
permeable film or membrane. - 1.enzyme coating
- 2.Nafion as coating is a breakthrough in
selectivity - b. electrochemical modification on working
electrode - applying high frequency triangular voltage
waveform onto electrodes in vitro. - c. adding chemicals to alter the composition of
the electrode - stearic acid modified carbon paste electrode
20How good sensitivity be made?
- Low sensitivity caused by biofoulingsuch as
protein deposition on electrode, impeding
electron transfer. - Solution to circumvent this problem
- electrode surface coated with permselective
coatings such as Nafion.
21 Less disturbance and good stability of
electrodes
- Carbon fiber as starting materials for working
electrodes. - stable, strong and compatible with various
modification. - Fabrication of microelectrodes
- carbon fiber electrode with small size up to
10-100µm. -
22what else can be done by in vivo CV?
- Voltammetry
- with brain slices
Jonathan A. Stamford, etal Analytical Chemistry
1996, (68) 359A-366A
23 What can be done in future
- Smaller electrodes needed (submicrometer ) to
study inferior structural and message
transmission information in single cell. - Development of integration and array of
microelectrodes technology can facilitate the
brain study in future. - Electrode modification will be further developed
in order to achieve better selectivity.
24We do help !!!