KENNEDY, Robert T.,YOON, Hyeun Joong,NGERNSUTIVORAKUL, Thitaphat,LEE, Woong Hee
申请号：
USUS2016/063930
公开号：
WO2017/100028A1
申请日:
2016.11.29
申请国别(地区):
WO
年份:
2017
代理人:
摘要：
Microdialysis sampling is an essential tool for in vivo neurochemical monitoring. Conventional dialysis probes are over 220 µm in diameter and have limited flexibility in design because they are made by assembly using preformed membranes. The probe size constrains spatial resolution and governs the amount of tissue damaged caused by probe insertion. To overcome these limitations, we have developed a method to microfabricate probes in Si that are 45 µm thick 180 µm wide. The probes contain a buried, U-shaped channel that is 30 µm deep 60 µm wide and terminates in ports for external connection. A 4 mm length of the probe is covered with a 5 µm thick nanoporous membrane. The membrane was microfabricated by deep reactive ion etching through a porous aluminum oxide layer. The microfabricated probe has cross-sectional area that is 79% less than that of the smallest conventional microdialysis probes. The probes yield 2-7% relative recovery at 100 nL/min perfusion rate for a variety of small molecules. The probe was successfully tested in vivo by sampling from the striatum of live rats. Fractions were collected at 20 min intervals (2 µL) before and after an injection of 5 mg/ kg, i.p amphetamine. Analysis of fractions by liquid chromatography-mass spectrometry revealed reliable detection of 13 neurochemicals, including dopamine and acetylcholine, at basal conditions. Amphetamine evoked a 43-fold rise in dopamine, a result nearly identical to a conventional dialysis probe in the same animal. The microfabricated probes have potential for sampling with higher spatial resolution and less tissue disruption than conventional probes. It may also be possible to add functionality to the probes by integrating other components, such as electrodes, optics, and additional channels.Léchantillonnage en microdialyse est un outil essentiel pour la surveillance dagents neurochimiques in vivo. Les sondes de dialyse classiques présentent un diamètre supérieur à 220 µm et sont dotées dune flexi
