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并行成像微流控细胞仪

Parallel imaging microfluidic cytometer.

作者信息

Ehrlich Daniel J, McKenna Brian K, Evans James G, Belkina Anna C, Denis Gerald V, Sherr David H, Cheung Man Ching

机构信息

Department of Biomedical Engineering, Boston University, Boston, Massachusetts, USA.

出版信息

Methods Cell Biol. 2011;102:49-75. doi: 10.1016/B978-0-12-374912-3.00003-1.

Abstract

By adding an additional degree of freedom from multichannel flow, the parallel microfluidic cytometer (PMC) combines some of the best features of fluorescence-activated flow cytometry (FCM) and microscope-based high-content screening (HCS). The PMC (i) lends itself to fast processing of large numbers of samples, (ii) adds a 1D imaging capability for intracellular localization assays (HCS), (iii) has a high rare-cell sensitivity, and (iv) has an unusual capability for time-synchronized sampling. An inability to practically handle large sample numbers has restricted applications of conventional flow cytometers and microscopes in combinatorial cell assays, network biology, and drug discovery. The PMC promises to relieve a bottleneck in these previously constrained applications. The PMC may also be a powerful tool for finding rare primary cells in the clinic. The multichannel architecture of current PMC prototypes allows 384 unique samples for a cell-based screen to be read out in ∼6-10 min, about 30 times the speed of most current FCM systems. In 1D intracellular imaging, the PMC can obtain protein localization using HCS marker strategies at many times for the sample throughput of charge-coupled device (CCD)-based microscopes or CCD-based single-channel flow cytometers. The PMC also permits the signal integration time to be varied over a larger range than is practical in conventional flow cytometers. The signal-to-noise advantages are useful, for example, in counting rare positive cells in the most difficult early stages of genome-wide screening. We review the status of parallel microfluidic cytometry and discuss some of the directions the new technology may take.

摘要

通过增加多通道流动的额外自由度,并行微流控细胞仪(PMC)结合了荧光激活流式细胞术(FCM)和基于显微镜的高内涵筛选(HCS)的一些最佳特性。PMC(i)适合快速处理大量样本,(ii)增加了用于细胞内定位分析(HCS)的一维成像能力,(iii)具有高稀有细胞灵敏度,以及(iv)具有时间同步采样的非凡能力。传统流式细胞仪和显微镜在组合细胞分析、网络生物学和药物发现中无法实际处理大量样本,这限制了它们的应用。PMC有望缓解这些先前受限应用中的瓶颈。PMC也可能是在临床中寻找稀有原代细胞的有力工具。当前PMC原型的多通道架构允许在约6 - 10分钟内读出用于基于细胞筛选的384个独特样本,速度约为大多数当前FCM系统的30倍。在一维细胞内成像中,PMC可以使用HCS标记策略获得蛋白质定位,其样本通量是基于电荷耦合器件(CCD)的显微镜或基于CCD的单通道流式细胞仪的许多倍。PMC还允许信号积分时间在比传统流式细胞仪更宽的范围内变化。例如,在全基因组筛选最困难的早期阶段对稀有阳性细胞进行计数时,信噪比优势很有用。我们回顾了并行微流控细胞术的现状,并讨论了这项新技术可能的一些发展方向。

相似文献

1
Parallel imaging microfluidic cytometer.并行成像微流控细胞仪
Methods Cell Biol. 2011;102:49-75. doi: 10.1016/B978-0-12-374912-3.00003-1.
3
Image-based cell-resolved screening assays in flow.流动状态下基于图像的细胞分辨筛选分析
Cytometry A. 2015 Jun;87(6):541-8. doi: 10.1002/cyto.a.22609. Epub 2014 Dec 16.

本文引用的文献

5
Microscopic imaging techniques for drug discovery.用于药物发现的显微成像技术。
Nat Rev Drug Discov. 2008 Jan;7(1):54-67. doi: 10.1038/nrd2446.
9
High-content screening moves to the front of the line.高内涵筛选走到了前列。
Drug Discov Today. 2006 Oct;11(19-20):889-94. doi: 10.1016/j.drudis.2006.08.015. Epub 2006 Sep 7.

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