Integrating impedance-based growth-rate monitoring into a microfluidic cell culture platform for live-cell microscopy

被引:24
作者
Chawla, Ketki [1 ]
Burgel, Sebastian C. [1 ]
Schmidt, Gregor W. [1 ]
Kaltenbach, Hans-Michael [2 ]
Rudolf, Fabian [2 ]
Frey, Olivier [1 ]
Hierlemann, Andreas [1 ]
机构
[1] Swiss Fed Inst Technol, Dept Biosyst Sci & Engn, Bio Engn Lab, Basel, Switzerland
[2] Swiss Fed Inst Technol, Dept Biosyst Sci & Engn, Computat Syst Biol Grp, Basel, Switzerland
来源
MICROSYSTEMS & NANOENGINEERING | 2018年 / 4卷
基金
瑞士国家科学基金会;
关键词
LONG-TERM; YEAST; FLOW; CHEMOSTAT; CYTOMETRY; BACTERIAL; DYNAMICS; CYCLE; CHIP;
D O I
10.1038/s41378-018-0006-5
中图分类号
TB3 [工程材料学];
学科分类号
0805 ; 080502 ;
摘要
Growth rate is a widely studied parameter for various cell-based biological studies. Growth rates of cell populations can be monitored in chemostats and micro-chemostats, where nutrients are continuously replenished. Here, we present an integrated microfluidic platform that enables long-term culturing of non-adherent cells as well as parallel and mutually independent continuous monitoring of (i) growth rates of cells by means of impedance measurements and of (ii) specific other cellular events by means of high-resolution optical or fluorescence microscopy. Yeast colonies were grown in a monolayer under culturing pads, which enabled high-resolution microscopy, as all cells were in the same focal plane. Upon cell growth and division, cells leaving the culturing area passed over a pair of electrodes and were counted through impedance measurements. The impedance data could then be used to directly determine the growth rates of the cells in the culturing area. The integration of multiple culturing chambers with sensing electrodes enabled multiplexed long-term monitoring of growth rates of different yeast strains in parallel. As a demonstration, we modulated the growth rates of engineered yeast strains using calcium. The results indicated that impedance measurements provide a label-free readout method to continuously monitor the changes in the growth rates of the cells without compromising high-resolution optical imaging of single cells.
引用
收藏
页数:12
相关论文
共 44 条
[31]   Single cell dielectric spectroscopy [J].
Morgan, Hywel ;
Sun, Tao ;
Holmes, David ;
Gawad, Shady ;
Green, Nicolas G. .
JOURNAL OF PHYSICS D-APPLIED PHYSICS, 2007, 40 (01) :61-70
[32]   Long-Term Single Cell Analysis of S. pombe on a Microfluidic Microchemostat Array [J].
Nobs, Jean-Bernard ;
Maerkl, Sebastian J. .
PLOS ONE, 2014, 9 (04)
[33]  
Rines DanielR., 2011, Cold Spring Harb Protoc, V9, P1026, DOI DOI 10.1101/PDB.T0P065482
[34]  
Schmidt GW, 2018, METHODS MOL BIOL, V1672, P537, DOI 10.1007/978-1-4939-7306-4_36
[35]   NIH Image to ImageJ: 25 years of image analysis [J].
Schneider, Caroline A. ;
Rasband, Wayne S. ;
Eliceiri, Kevin W. .
NATURE METHODS, 2012, 9 (07) :671-675
[36]   Coupling among growth rate response, metabolic cycle, and cell division cycle in yeast [J].
Slavov, Nikolai ;
Botstein, David .
MOLECULAR BIOLOGY OF THE CELL, 2011, 22 (12) :1997-2009
[37]   Positional dependence of particles in microfludic impedance cytometry [J].
Spencer, Daniel ;
Morgan, Hywel .
LAB ON A CHIP, 2011, 11 (07) :1234-1239
[38]   Single-cell microfluidic impedance cytometry: a review [J].
Sun, Tao ;
Morgan, Hywel .
MICROFLUIDICS AND NANOFLUIDICS, 2010, 8 (04) :423-443
[39]   Integrated systems for rapid point of care (PoC) blood cell analysis [J].
van Berkel, Cees ;
Gwyer, James D. ;
Deane, Steve ;
Green, Nicolas ;
Holloway, Judith ;
Hollis, Veronica ;
Morgan, Hywel .
LAB ON A CHIP, 2011, 11 (07) :1249-1255
[40]   OPINION Metabolism, cell growth and the bacterial cell cycle [J].
Wang, Jue D. ;
Levin, Petra A. .
NATURE REVIEWS MICROBIOLOGY, 2009, 7 (11) :822-827