Nanofluidic platform for single mitochondria analysis using fluorescence microscopy.
Nanofluidic platform for single mitochondria analysis using fluorescence microscopy.
复制标题
使用荧光显微镜进行单个线粒体分析的纳流体平台。
DOI：
10.1021/ac4010088
复制
发表时间：
2013
影响因子：
7.4
通讯作者：
Burke,PeterJ
中科院分区：
文献类型：
--
作者：
Zand,Katayoun;Pham,Ted;DavilaJr,Antonio;Wallace,DouglasC;Burke,PeterJ
使用横截面为500 nm × 2 μm的PDMS中的纳米流体通道，我们展示了单个分离线粒体的捕获和询问。荧光标记显示线粒体固定在沿着通道的离散位置。用不同的电位敏感染料（JC-1和TMRM）询问线粒体膜电位表明被捕获的线粒体在呼吸缓冲液中是至关重要的。在单个线粒体水平上可以观察到膜电位的波动。可以将各种化学挑战递送到纳米流体系统中的每个单独的微扰子。作为样品证明，在将OXPHOS底物引入纳米流体通道中时，观察到膜电位的增加。将Ca 2+引入纳米通道诱导线粒体膜透化（MMP），导致在单个线粒体水平观察到的去极化。可以设想使用该技术在癌症生物学、干细胞生物学、细胞凋亡研究和高通量功能代谢组学研究中的各种应用。
Using nanofluidic channels in PDMS of cross section 500 nm × 2 μm, we demonstrate the trapping and interrogation of individual, isolated mitochondria. Fluorescence labeling demonstrates the immobilization of mitochondria at discrete locations along the channel. Interrogation of mitochondrial membrane potential with different potential sensitive dyes (JC-1 and TMRM) indicates the trapped mitochondria are vital in the respiration buffer. Fluctuations of the membrane potential can be observed at the single mitochondrial level. A variety of chemical challenges can be delivered to each individual mitochondrion in the nanofluidic system. As sample demonstrations, increases in the membrane potential are seen upon introduction of OXPHOS substrates into the nanofluidic channel. Introduction of Ca2+into the nanochannels induces mitochondrial membrane permeabilization (MMP), leading to depolarization, observed at the single mitochondrial level. A variety of applications in cancer biology, stem cell biology, apoptosis studies, and high throughput functional metabolomics studies can be envisioned using this technology.