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一种用于轻松收集和质谱表征分泌型神经肽的神经元-毛细血管平台。

A neuron-in-capillary platform for facile collection and mass spectrometric characterization of a secreted neuropeptide.

作者信息

Lee Chang Young, Fan Yi, Rubakhin Stanislav S, Yoon Sook, Sweedler Jonathan V

机构信息

Department of Chemistry and the Beckman Institute, University of Illinois, Urbana, IL 61801, USA.

School of Energy and Chemical Engineering, School of Life Sciences, Ulsan National Institute of Science and Technology (UNIST), Ulsan 44919, Republic of Korea.

出版信息

Sci Rep. 2016 Jun 1;6:26940. doi: 10.1038/srep26940.

Abstract

The integration of microfluidic devices-which efficiently handle small liquid volumes-with separations/mass spectrometry (MS) is an effective approach for profiling the neurochemistry occurring in selected neurons. Interfacing the microfluidic cell culture to the mass spectrometer is challenging because of geometric and scaling issues. Here we demonstrate the hyphenation of a neuron-in-capillary platform to a solid phase extraction device and off-line MS. A primary neuronal culture of Aplysia californica neurons was established directly inside a cylindrical polyimide capillary. The approach also uses a particle-embedded monolith to condition neuropeptide releasates collected from several Aplysia neurons cultured in the capillary, with the subsequent characterization of released peptides via MS. This system presents a number of advances compared to more traditional microfluidic devices fabricated with polydimethylsiloxane. These include low cost, easy access to cell culture, rigidity, ease of transport, and minimal fluid handling. The cylindrical geometry of the platform allows convenient interface with a wide range of analytical tools that utilize capillary columns.

摘要

将能够高效处理少量液体的微流控装置与分离/质谱联用是描绘特定神经元中发生的神经化学特征的有效方法。由于几何形状和缩放问题,将微流控细胞培养与质谱仪连接具有挑战性。在此,我们展示了将毛细管内神经元平台与固相萃取装置和离线质谱联用。在圆柱形聚酰亚胺毛细管内直接建立了加州海兔神经元的原代培养。该方法还使用了颗粒嵌入整体柱来处理从毛细管中培养的多个海兔神经元收集的神经肽释放物,随后通过质谱对释放的肽进行表征。与用聚二甲基硅氧烷制造的更传统的微流控装置相比,该系统有许多进步。这些进步包括低成本、易于进行细胞培养、刚性、易于运输以及最少的流体处理。该平台的圆柱形几何形状便于与广泛使用毛细管柱的分析工具进行连接。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6142/4887886/a43413f0c26a/srep26940-f1.jpg

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