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微透析探头可用来探测在体和离体神经传递素和其它代谢物。多数情况下,这些探头被植入脑组织中,或者用于血液微透析中。
可以使用流动体系进行微透析实验,在流动体系中,溶液被泵流过微透析探头以富集代谢物,然后这些代谢物被填进探测器。在多数情况下使用电化学方法探测,因其高度的灵敏性,但从理论上讲也可以使用任何检测器例如荧光检测器或吸光度检测器。
PowerChrom和e-corder连用兼容绝大多数HPLC检测器,这些检测器包括电化学检测器可用于微透析的研究。以下公司生产电化学检测器: |
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Antec Leyden, 'Intro'
电化学检测器 |
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BAS
(Bioanalytical Systems),
LC-4C电化学检测器 |
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Eicom |
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ESA,
Model 5200A电化学检测器 |
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GBC
Scientific, LC1260电化学检测器 |
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e-corder系统可以使用Chart软件或PowerChrom软件来记录数据。PowerChrom
280和285系统使用PowerChrom软件和专用的数据记录硬件。(PowerChrom
285系统甚至具有内置的PC计算机,预装Window XP和PowerChrom色谱工作站软件)。
有些情况下,可以在微透析探头内部进行电化学检测。请参考: |
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Sycopel微透析电极 |
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这种电极有几种不同的配置型号,有离体使用的,也有在体使用的。可以将溶液通过泵驱动流经微透析电极,或者在静止的溶液中使用微透析电极。这些电极连接到: |
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Sycopel生物传感器驱动器,
或者 |
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微电流恒电位仪 |
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在静止溶液中使用微透析电极时,通常使用e-corder与Chart软件。如果溶液被泵流经微透析电极,那么可以使用e-corder主机和Chart软件,或者PowerChrom
280系统记录数据信号。 |
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使用PowerChrom系统研究微透析的实例文献 |
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与脑桥诱导缓驰相关的抑制氨基酸释放的变化:在体微透析研究。
Changes in inhibitory amino acid
release linked to pontine-induced atonia:
An in vivo microdialysis study. |
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| T
Kodama, Y-Y Lai and J M Siegel, Journal of Neuroscience, 23,
1548-1554, 2003. |
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灌流液中的氨基酸浓度由HPLC(EDT-300;
Eicom)测定。HPLC带荧光检测器(在340 nm激励,440 nm发射),用PowerChrom系统进行量化。
The concentration of amino
acid in the perfusate was determined by HPLC (EDT-300;
Eicom) witha
fluorescence detector (excitation at 340 nm, emission at
440 nm) and quantified with a PowerChrom system. |
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用微透析监测自由活动的老鼠脑皮层中葡萄糖和乳酸盐的动态变化。
Dynamic Changes in Glucose and
Lactate in the Cortex of the Freely Moving Rat Monitored Using
Microdialysis. |
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| D. A.
Jones, J. Ros, H. Landolt, M. Fillenz, and M. G. Boutelle,
Journal of Neurochemistry, 75, 1703-1708, 2000. |
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应用双重在线化验方法在线分析透析液中的葡萄糖和乳酸盐。该化验方法基于固定酶床/二茂铁媒介探测技术。用HPLC的泵将包含羧酸二茂铁媒介的缓冲液(pH 7)以0.6 mL/分钟的速率连续泵入阀中。每个化验系统包含一个小的酶床反应器(1
mmx
20 mm,包含葡萄糖氧化酶和山葵过氧化酶,或乳酸盐氧化酶和山葵过氧化酶)和一个下游玻璃碳幅流式电极固定电位于-100
mV相对于Ag/AgCl参比电极。阀的开关和数据的采集使用PowerChrom系统进行。
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. |
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伏隔核中的局部阿尔法-金环蛇毒素敏感烟碱感受器在体调制烟碱诱发多巴胺分泌
Local alpha-bungarotoxin-sensitive
nicotinic receptors in the nucleus accumbens modulate
nicotine-stimulated dopamine secretion in vivo. |
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| Y. Fu,
S.G. Matta, W. Gao, and B.M. Sharp, Neuroscience, 101, 369-375,
2000. |
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NAcc中由系统烟碱对多巴胺(DA)分泌的诱发被阿尔法-金环蛇毒素有选择性地抑制,或者由甲基牛扁碱直接抑制到此区域,此处盐酸-3-甲基氨基异樟脑烷无效。多巴胺分泌的在体微透析和感受器在腹鳍和伏隔核及中脑腹鳍盖处被阻塞。微透析用ESA
Coulochem II
5200A电化学检测器分析,带高灵敏分析池。电化学检测在220 mV电位处10 nA电流增益执行。在这些条件下,DA的检出限为100 fg每次注射。色谱数据用PowerChrom系统采集和分析。
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 ata
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. |
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草胺磷通过老鼠小脑中的N-甲基D-天门冬氨酸盐感受器刺激一氧化氮的生成
Glufosinate
ammonium stimulates nitric oxide production through N-methyl D-aspartate
receptors in rat cerebellum. |
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| Toshio
Nakaki, Akira Mishima, Eiji Suzuki, Futoshi Shintani, and Tomoko
Fujii, Neuroscience Letters, 290, 209-212, 2000. |
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生成的一氧化氮由其非酶氧化产物亚硝酸盐/硝酸盐来定量表示。亚硝酸盐/硝酸盐由配置有液相色谱和Griess反应(ENO-100,
Eicom, Kyoto,
日本)的反应器的自动系统来定量,在反应器中,用一根镉柱将硝酸盐还原成亚硝酸盐。反应物的吸光率由一个经流式的分光光度计(NOD-10,Eicom)在540 nm测定。用PowerChrom系统采集和分析数据。
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 witha
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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