Simultaneous detection of the release of glutamate and nitric oxide from adherently growing cells using an array of glutamate and nitric oxide selective electrodes.
The simultaneous detection of nitric oxide and glutamate using an array of individually addressable electrodes, in which the individual electrodes in the array were suitably modified with a highly sensitive nitric oxide sensing chemistry or a glutamate oxidase/redox hydrogel-based glutamate biosensor is presented. In a sequence of modification steps one of the electrodes was covered first with a positively charged Ni porphyrin entrapped into a negatively charged electrodeposition paint followed by the manual modification of the second working electrode by a bienzyme sensor architecture based on crosslinked redox hydrogels with entrapped peroxidase and glutamate oxidase. Adherently growing C6-glioma cells were grown on membrane inserts and placed in close distance to the modified sensor surfaces. The current responses recorded at each electrode after stimulation of glutamate and NO release by means of K+ and bradykinin clearly demonstrate the ability of the individual electrode in the array to detect the analyte towards which its sensitivity and selectivity was targeted without interference from the neighbouring electrode or other analytes present in the test mixture.
Castillo
Jaime
J
Department of Biotechnology, Lund University, S-22100 Lund, Sweden.
Isik
Sonnur
S
Blöchl
Andrea
A
Pereira-Rodrigues
Nazaré
N
Bedioui
Fethi
F
Csöregi
Elizabeth
E
Schuhmann
Wolfgang
W
Oni
Joshua
J
D000818:Animals; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D002448:Cell Adhesion; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods; D045744:Cell Line, Tumor; D018914:Cells, Immobilized; D020099:Coated Materials, Biocompatible / Q000737:chemistry; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005910:Glioma / Q000378:metabolism*; D018698:Glutamic Acid / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism*; D008839:Microelectrodes; D009569:Nitric Oxide / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism*; D051381:Rats; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D020099:Coated Materials, Biocompatible; D009569:Nitric Oxide; D018698:Glutamic Acid