Improved electrochemical detection of biogenic amines in Aplysia using base-hydrolyzed cellulose-coated carbon fiber microelectrodes.
A major challenge with electrochemical techniques in vivo, using carbon-fiber microelectrodes, is to achieve sufficient sensitivity to detect the low concentrations of transmitters released by neurons. In particular, when an electrode is inserted into living tissue, its sensitivity is usually substantially decreased as a result of the degradation of the carbon surface by proteins. Here, we show that this decrease can be significantly attenuated by coating the electrode with cellulose acetate. The cellulose film offers a steric barrier that prevents macromolecules from diffusing to the carbon surface and its porosity can be progressively increased by controlled hydrolysis. We compared different cellulose-coated electrodes, either non-hydrolysed or hydrolyzed, in 0.08 N KOH for 10-30 min. We found that dopamine and serotonin detection was blocked by non-hydrolysed cellulose films, but that hydrolysis restored optimal detection similar to uncoated electrodes. Moreover, cellulose films (hydrolyzed for 20 min) significantly diminished electrode degradation in vivo and allowed reliable detection of fast concentration changes with <0.5 s delay, compared to uncoated electrodes. Finally, the sensitivity to endogenous 5-HT release in Aplysia central nervous system was more than doubled with these electrodes. We conclude that the optimal hydrolysis time of cellulose-coated electrodes is approximately 20 min with our protocol and carbon fiber electrodes prepared with this method offer improved sensitivity for the detection of biogenic amines.
Marinesco
Stéphane
S
Department of Neurobiology and Behavior and Center for the Neurobiology of Learning and Memory, University of California, Irvine, CA 92697-4550, USA. smarines@uci.edu
Carew
Thomas J
TJ
D000818:Animals; D001048:Aplysia / Q000737:chemistry* / Q000166:cytology; D015306:Biogenic Monoamines / Q000032:analysis*; D002244:Carbon; D000077482:Carbon Fiber; D002482:Cellulose; D002490:Central Nervous System / Q000737:chemistry* / Q000166:cytology; D004298:Dopamine / Q000032:analysis; D004563:Electrochemistry / Q000295:instrumentation / Q000379:methods*; D004566:Electrodes / Q000592:standards* / Q000639:trends; D005465:Fluorocarbon Polymers; D006868:Hydrolysis; D009474:Neurons / Q000737:chemistry* / Q000166:cytology; D011930:Reaction Time; D012701:Serotonin / Q000032:analysis
D015306:Biogenic Monoamines; D000077482:Carbon Fiber; D005465:Fluorocarbon Polymers; D012701:Serotonin; C040402:perfluorosulfonic acid; D002244:Carbon; D002482:Cellulose; D004298:Dopamine