Microfluidic single-cell cultivation chip with controllable immobilization and selective release of yeast cells

Zhu, Zhen; Frey, Olivier; Ottoz, Diana S. M.; Rudolf, Fabian; Hierlemann, Andreas

doi:10.1039/c2lc20911j

Cited by 69 publications

(69 citation statements)

References 33 publications

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“…Several other methods have also been coupled with microfluidics for cell immobilization and conducting controlled, complete cell assays. Flow-based active cell trapping by using control valves [14,15], non-invasive optical trapping [16][17][18], dielectrophoresis [19][20][21], surface chemistry modification techniques [22,23], arrays of physical barriers [24], cell trapping by negative pressure [25,26], and hydrodynamic methods [27][28][29] are some of these successfully established techniques.…”

Section: Introductionmentioning

confidence: 99%

Hydrodynamic Cell Trapping for High Throughput Single-Cell Applications

et al. 2013

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Abstract:The possibility to conduct complete cell assays under a precisely controlled environment while consuming minor amounts of chemicals and precious drugs have made microfluidics an interesting candidate for quantitative single-cell studies. Here, we present an application-specific microfluidic device, cellcomb, capable of conducting high-throughput single-cell experiments. The system employs pure hydrodynamic forces for easy cell trapping and is readily fabricated in polydimethylsiloxane (PDMS) using soft lithography techniques. The cell-trapping array consists of V-shaped pockets designed to accommodate up to six Saccharomyces cerevisiae (yeast cells) with the average diameter of 4 μm. We used this platform to monitor the impact of flow rate modulation on the arsenite (As(III)) uptake in yeast. Redistribution of a green fluorescent protein (GFP)-tagged version of the heat shock protein Hsp104 was followed over time as read out. Results showed a clear reverse correlation between the arsenite uptake and three different adjusted low = 25 nL min −1 , moderate = 50 nL min −1 , and high = 100 nL min −1 flow rates. We consider the presented device as the first building block of a future integrated application-specific cell-trapping array that can be used to conduct complete single cell experiments on different cell types.

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Section: Introductionmentioning

confidence: 99%

Hydrodynamic Cell Trapping for High Throughput Single-Cell Applications

et al. 2013

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“…In this work, we have integrated EIS into a microfluidic device for single-cell analysis. The concept of single-cell manipulation has been characterized and validated previously [35,36] and has been adapted and extended with specifically designed electrodes. The device features reliable immobilization and the possibility to cultivate of single cells under controlled environmental conditions while performing real-time impedance measurements of the immobilized cells.…”

Section: Introductionmentioning

confidence: 99%

Real-time monitoring of immobilized single yeast cells through multifrequency electrical impedance spectroscopy

et al. 2014

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We present a microfluidic device, which enables single cells to be reliably trapped and cultivated while simultaneously being monitored by means of multifrequency electrical impedance spectroscopy (EIS) in the frequency range of 10 kHz-10 MHz. Polystyrene beads were employed to characterize the EIS performance inside the microfluidic device. The results demonstrate that EIS yields a low coefficient of variation in measuring the diameters of captured beads (~0.13 %). Budding yeast, Saccharomyces cerevisiae, was afterwards used as model organism. Single yeast cells were immobilized and measured by means of EIS. The bud growth was monitored through EIS at a temporal resolution of 1 min. The size increment of the bud, which is difficult to determine optically within a short time period, can be clearly detected through EIS signals. The impedance measurements also reflect the changes in position or motion of single yeast cells in the trap. By analyzing the multifrequency EIS data, cell motion could be qualitatively discerned from bud growth. The results demonstrate that single-cell EIS can be used to monitor cell growth, while also detecting potential cell motion in real-time and label-free approach, and that EIS constitutes a sensitive tool for dynamic single-cell analysis.

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“…The immobilization realized by both approaches is a statistical process and does not allow for a specific cell to be taken out of the flow or released again. More active control over the immobilization was achieved by applying negative pressure in order to pull cells into narrow channels arranged along the channel walls [25][26][27][28]. Throughput is typically decreased by better spatial control over the single cells.…”

Section: Introductionmentioning

confidence: 99%

Polydimethylsiloxane (PDMS) Sub-Micron Traps for Single-Cell Analysis of Bacteria

et al. 2013

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Abstract:Microfluidics has become an essential tool in single-cell analysis assays for gaining more accurate insights into cell behavior. Various microfluidics methods have been introduced facilitating single-cell analysis of a broad range of cell types. However, the study of prokaryotic cells such as Escherichia coli and others still faces the challenge of achieving proper single-cell immobilization simply due to their small size and often fast growth rates. Recently, new approaches were presented to investigate bacteria growing in monolayers and single-cell tracks under environmental control. This allows for high-resolution time-lapse observation of cell proliferation, cell morphology and fluorescence-coupled bioreporters. Inside microcolonies, interactions between nearby cells are likely and may cause interference during perturbation studies. In this paper, we present a microfluidic device containing hundred sub-micron sized trapping barrier structures for single E. coli cells. Descendant cells are rapidly washed away as well as components secreted by growing cells. Experiments show excellent growth rates, indicating high cell viability. Analyses of elongation and growth rates as well as morphology were successfully performed. This device will find application in prokaryotic single-cell studies under constant environment where by-product interference is undesired.

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Microfluidic single-cell cultivation chip with controllable immobilization and selective release of yeast cells

Cited by 69 publications

References 33 publications

Hydrodynamic Cell Trapping for High Throughput Single-Cell Applications

Hydrodynamic Cell Trapping for High Throughput Single-Cell Applications

Real-time monitoring of immobilized single yeast cells through multifrequency electrical impedance spectroscopy

Polydimethylsiloxane (PDMS) Sub-Micron Traps for Single-Cell Analysis of Bacteria

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