Microfluidics-integrated time-lapse imaging for analysis of cellular dynamics.
Microfluidics-integrated time-lapse imaging for analysis of cellular dynamics.
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DOI：
10.1039/b923699f
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发表时间：
2010-06
期刊：
影响因子：
--
通讯作者：
Shah JV
中科院分区：
文献类型：
--
作者：
Albrecht DR;Underhill GH;Resnikoff J;Mendelson A;Bhatia SN;Shah JV
最近的创新，使观察表型在高时空分辨率的调节细胞反应的机制的理解得到了加强。诸如微流体技术的技术已经寻求扩大这些方法的通量，尽管与先进成像策略的同化受到限制。在这里，我们描述了配对的高分辨率延时成像与微流体多路复用的细胞动力学分析，利用设计选择的简易制造和操作，并与显微镜仪器集成。这种模块化的中等通量平台通过使用保守的96孔、~6 × 5 mm 2成像区域以及根据研究中的细胞类型和微环境数量选择的可变输入/输出通道设计，能够在高数值孔径下（通过油浸）对活细胞进行长期成像。在该系统的验证中，我们研究了细胞周期进展的基本特征，包括有丝分裂动力学和纺锤体取向动力学，通过高分辨率的平行分析模型细胞系进行抗有丝分裂剂。我们还探索了小鼠胚胎干细胞的自我更新动力学，并证明了动态评估和操纵干细胞增殖，检测罕见细胞事件和测量扩展时间尺度相关性的能力。我们实现了>900个细胞/实验的实验通量，每个细胞在>40×放大倍数下观察长达120小时。总的来说，这些研究说明了通过整合一个简单的、模块化的、基于微流体的成像平台来探测细胞功能并产生时间和空间动态信息的能力。
An understanding of the mechanisms regulating cellular responses has recently been augmented by innovations enabling the observation of phenotypes at high spatiotemporal resolution. Technologies such as microfluidics have sought to expand the throughput of these methods, although assimilation with advanced imaging strategies has been limited. Here, we describe the pairing of high resolution time-lapse imaging with microfluidic multiplexing for the analysis of cellular dynamics, utilizing a design selected for facile fabrication and operation, and integration with microscopy instrumentation. This modular, medium-throughput platform enables the long-term imaging of living cells at high numerical aperture (via oil immersion) by using a conserved 96-well, ~6 × 5 mm2 imaging area with a variable input/output channel design chosen for the number of cell types and microenvironments under investigation. In the validation of this system, we examined fundamental features of cell cycle progression, including mitotic kinetics and spindle orientation dynamics, through the high-resolution parallel analysis of model cell lines subjected to anti-mitotic agents. We additionally explored the self-renewal kinetics of mouse embryonic stem cells, and demonstrate the ability to dynamically assess and manipulate stem cell proliferation, detect rare cell events, and measure extended time-scale correlations. We achieved an experimental throughput of >900 cells/experiment, each observed at >40× magnification for up to 120 h. Overall, these studies illustrate the capacity to probe cellular functions and yield dynamic information in time and space through the integration of a simple, modular, microfluidics-based imaging platform.
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影响因子：
64.8
作者：
通讯作者：
--
影响因子：
12.3
作者：
Carpenter AE;Jones TR;Lamprecht MR;Clarke C;Kang IH;Friman O;Guertin DA;Chang JH;Lindquist RA;Moffat J;Golland P;Sabatini DM
通讯作者：
Sabatini DM
影响因子：
48
作者：
Loo, Lit-Hsin;Wu, Lani F.;Altschuler, Steven J.
通讯作者：
Altschuler, Steven J.
影响因子：
3.4
作者：
Botvinick, EL;Venugopalan, V;Berns, MW
通讯作者：
Berns, MW
影响因子：
4
作者：
Alvarez, D;Novac, O;Zannis-Hadjopoulos, M
通讯作者：
Zannis-Hadjopoulos, M