Hi I am recently learning to use fast-scan cyclic voltammetry to measure dopamine (DA) in striatum in vivo, but had no luck. Our system: Chem-Clamp (Dagan Corp) with the headstage gain modified by ourselves. The carbon fiber microelectrode (CFM) was hand-made using glassy carbon fiber (monofilament) purchased from Specialty Materials (Lowell, MA) http://www.specmaterials.com/carbonmonofilament.htm.
We stimulated the MFB with an iso-flex stimulator using a twisted bipolar needle-style stimulating electrode (home-made). The whole system was grounded by an Ag/AgCl wire
We could get reasonable voltammogram curves similar to those in literatures when calibrating the CFM with exogeneous DA at 2 micromolar-and-up range, but had no luck in detecting evoked DA release in vivo, no matter how we tried.
I had suspected that we did not stimulat on the right spot. So we tried various different stimulating and recording coordinates for both the stimulating electrode and CFM, but still had no luck. The most commonly used cooridinates are: AP2.5, ML 2.5, DL 4.5 for CFM; AP -4.6, ML 1.3, DL 7~9 for stimulation.
We had tried so many times, but still could not get it right. I am now at my wits' end. So I am hoping if someone could help me to set thing right. I have a few specific questions:
1. Is the glassy carbon an appropriate material for use of FSCV in measuring dopamine?
2. Is my in-vitro voltammogram curve looking correct?
3. Why couldn't we detect the evoked DA release in vivo ? Did we miss or do anything wrong? Coordinates problems? Any suggestions?
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