Continuous monitoring of L-glutamate released from cultured nerve cells by an online sensor coupled with microcapillary sampling

Osamu Niwa, Tsutomu Horiuchi, Keiichi Torimitsu

Research output: Contribution to journalArticlepeer-review

51 Citations (Scopus)

Abstract

A small volume L glutamate online sensor was developed in order to monitor changes in the local concentration of L-glutamate released from cultured nerve cells. Syringe pump in the suction mode is used to sample extracellular fluid continuously from a glass micro-capillary and the concentration of L-glutamate can be determined by using a glassy carbon (GC) electrode modified with an Os-polyvinylpyridine mediator bottom film containing horseradish peroxidase and a bovine serum albumin top layer containing L-glutamate oxidase. The overall efficiency of L-glutamate detection with a sensor is 71% under optimum conditions due to an efficient enzymatic reaction at the modified electrode in the thin layer radial flow cell. As a result, we achieved a detection limit of 7-15 nM and a linear range of 50 nM to 10 μM. In an in vitro experiment, the extracellular fluid near a particular nerve cell can be sampled with this micro-pipet and continuously introduced into the modified GC electrode in the radial flow cell via suction provided by a syringe pump. The nerve cells are stimulated by KCl in a glass capillary and the L-glutamate concentration change can be monitored by changing the distance between the sampling piper and the nerve cells.

Original languageEnglish
Pages (from-to)311-319
Number of pages9
JournalBiosensors and Bioelectronics
Volume12
Issue number4
DOIs
Publication statusPublished - 1997 Jun 19
Externally publishedYes

Keywords

  • L-glutamate sensor
  • capillary
  • enzyme
  • nerve cell
  • online

ASJC Scopus subject areas

  • Biotechnology
  • Biophysics
  • Biomedical Engineering
  • Electrochemistry

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