マイクロダイアリシス
脳内マイクロダイアリシスプローブを使用して、インビボ、インビトロで神経伝達物質やその代謝物の検出ができます。通常はダイアリシスプローブを脳組織に埋め込んだり、血管透析用として使用されています。
マイクロダイアリシスはフローシステムで行こなわれます。透析溶液をプローブに流し透析されて溶出した物質を含む透析溶液をHPLCで分離したり直接検出器で分析します。この種の分析ではHPLCを用いた電気化学検出法が一般的ですが、蛍光法や吸光度等の検出器も利用されています。
PowerChrom や e-corder がマイクロダイアリシス分析に使用されています。
主な電気化学検出器のメーカ:
- Antec Leyden, 'Intro' 電気化学検出器
- BAS(Bioanalytical Systems), LC-4C 電気化学検出器
- Eicom
- ESA, Model 5200A 電気化学検出器
- GBC Scientific, LC1260 電気化学検出器
e-corder ではChart ソフトウェアか PowerChrom ソフトウェアを使ってデータを記録します。PowerChrom システム ではクロマトグラム専用のPowerChrom ソフトウェアを使います。
マイクロダイアリシスプローブ内に電極を装着したタイプのプローブもあります:
この電極には幾種類もの形状が提供されています。透析液は通常潅流しますが、電極で測定する時はフローを停止します。使用する測定器:
- EA362 デュアルピコスタット
- EA164 ４連ポテンショスタット
測定データの記録は e-corder で Chart か PowerChrom ソフトウェアを使います。
引用文献
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Changes in inhibitory amino acid release linked to pontine-induced atonia: An in vivo microdialysis study. T Kodama, Y-Y Lai and J M Siegel, Journal of Neuroscience, 23, 1548-1554, 2003.
"The concentration of amino acid in the perfusate was determined by HPLC (EDT-300; Eicom) with a fluorescence detector (excitation at 340 nm, emission at 440 nm) and quantified with a PowerChrom system." -
Dynamic Changes in Glucose and Lactate in the Cortex of the Freely Moving Rat Monitored Using Microdialysis. D. A. Jones, J. Ros, H. Landolt, M. Fillenz, and M. G. Boutelle, Journal of Neurochemistry, 75, 1703-1708, 2000.
"Dialysate was analysed on-line for glucose and lactate using a dual on-line assay. The assay is based on immobilised enzyme bed/ferrocene-mediated detection technology. A buffer (pH 7) containing the ferrocene monocarboxylic acid mediator is continuously pumped into the valve at a flow rate of 0.6 mL/min using an HPLC pump. Each assay system consists of a small enzyme bed reactor (1 mm x 20 mm, containing either glucose oxidase and horseradish peroxidase or lactate oxidase and horseradish peroxidase) and a downstream glassy carbon radial flow electrode held at -100 mV relative to an Ag/AgCl reference electrode. Valve switching and data collection were done using PowerChrom software." -
Local alpha-bungarotoxin-sensitive nicotinic receptors in the nucleus accumbens modulate nicotine-stimulated dopamine secretion in vivo. Y. Fu, S.G. Matta, W. Gao, and B.M. Sharp, Neuroscience, 101, 369-375, 2000.
"Stimulation of dopamine (DA) secretion in the NAcc by systemic nicotine was inhibited by selectively administering a-bungarotoxin or methyllycaconitine directly into this region, whereas mecamylamine was ineffective. In vivo microdialysis of accumbal dopamine secretion and receptor antagonist blockade in both the ventral striatal nucleus accumbens and the midbrain ventral tegmental area were used. Microdialysates were analysed by an ESA Coulochem II 5200A electrochemical detector with an ESA 5041 high-sensitivity analytical cell. Electrochemical detection was performed at a potential of 220 mV with the current gain at 10 nA. Under these conditions, the limit of detection for DA was 100 fg per injection. Chromatographic data were collected and analysed with a PowerChrom system." -
Glufosinate ammonium stimulates nitric oxide production through N-methyl D-aspartate receptors in rat cerebellum. Toshio Nakaki, Akira Mishima, Eiji Suzuki, Futoshi Shintani, and Tomoko Fujii, Neuroscience Letters, 290, 209-212, 2000.
"Nitric oxide production was estimated by the quantitation of nitrite/nitrate, the non-enzymatic oxidative products of nitric oxide. The amount of nitrite/nitrate was quantitated with an automatic system equipped with a liquid chromatograph and a reactor for the Griess reaction (ENO-100, Eicom, Kyoto, Japan), whereby nitrate was reduced to nitrite with a cadmium column. The absorbance of the reactant was measured at 540 nm with a flow-through spectrophotometer (NOD-10, Eicom). Data were collected and analysed with a Powerchrom system."
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This page was last modified on: 08 Sep 2020 18:59:47
