The VersaLive platform enables pipette-compatible microfluidic mammalian cell culture for versatile applications

Microfluidic-based cell culture allows for precise spatio-temporal regulation of microenvironment, live cell imaging and better recapitulation of physiological conditions, while minimizing reagents consumption. Despite their usefulness, most microfluidic systems are designed with one specific application in mind and require specialized equipment and expertise for their operation. All these requirements prevent microfluidic- based cell culture to be widely adopted. Here, we designed and implemented a versatile and easy-to-use perfusion cell culture microfluidic platform for multiple application (VersaLive) requiring only standard pipettes. Here, we showcase the multiple uses of VersaLive (e.g., time-lapse live cell imaging, immunostaining, cell recovery, cell lysis) on mammalian cell lines and primary cells. VersaLive can replace standard cell culture formats in several applications, thus decreasing costs and increasing reproducibility across laboratories. The layout, documentation and protocols are open-source and available online at https://versalive.tigem.it/.

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Time-Lapse, Live-Cell Imaging of Potassium Efflux in Cell Exposed to Nanosecond Pulsed Electric Fields

Biophysical Journal ◽

10.1016/j.bpj.2020.11.1493 ◽

2021 ◽

Vol 120 (3) ◽

pp. 223a

Author(s):

Flavia Mazzarda ◽

Esin B. Sozer ◽

Julia L. Pittaluga ◽

Claudia Muratori ◽

P. Thomas Vernier

Keyword(s):

Electric Fields ◽

Live Cell Imaging ◽

Cell Imaging ◽

Pulsed Electric Fields ◽

Time Lapse ◽

Live Cell ◽

Potassium Efflux ◽

Nanosecond Pulsed Electric Fields

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Dye selection for live cell imaging of intact siRNA

Biological Chemistry ◽

10.1515/bc-2011-256 ◽

2012 ◽

Vol 393 (1-2) ◽

pp. 23-35 ◽

Cited By ~ 10

Author(s):

Markus Hirsch ◽

Dennis Strand ◽

Mark Helm

Keyword(s):

Live Cell Imaging ◽

Cell Imaging ◽

Fluorescent Dyes ◽

Resonance Energy Transfer ◽

Resonance Energy ◽

Time Lapse ◽

Live Cell ◽

High Yield ◽

Ph Variation ◽

Rnai Efficiency

Abstract Investigations into the fate of small interfering RNA (siRNA) after transfection may unravel new ways to improve RNA interference (RNAi) efficiency. Because intracellular degradation of RNA may prevent reliable observation of fluorescence-labeled siRNA, new tools for fluorescence microscopy are warranted to cover the considerable duration of the RNAi effect. Here, the characterization and application of new fluorescence resonance energy transfer (FRET) dye pairs for sensing the integrity of duplex siRNA is reported, which allows an assessment of the degradation status of an siRNA cell population by live cell imaging. A panel of high-yield fluorescent dyes has been investigated for their suitability as FRET pairs for the investigation of RNA inside the cell. Nine dyes in 13 FRET pairs were evaluated based on the performance in assays of photostability, cross-excitation, bleed-through, as well as on quantified changes of fluorescence as a consequence of, e.g., RNA strand hybridization and pH variation. The Atto488/Atto590 FRET pair has been applied to live cell imaging, and has revealed first aspects of unusual trafficking of intact siRNA. A time-lapse study showed highly dynamic movement of siRNA in large perinuclear structures. These and the resulting optimized FRET labeled siRNA are expected to have significant impact on future observations of labeled RNAs in living cells.

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Live Cell Imaging of Bacillus subtilis and Streptococcus pneumoniae using Automated Time-lapse Microscopy

Journal of Visualized Experiments ◽

10.3791/3145 ◽

2011 ◽

Cited By ~ 35

Author(s):

Imke G. de Jong ◽

Katrin Beilharz ◽

Oscar P. Kuipers ◽

Jan- Willem Veening

Keyword(s):

Bacillus Subtilis ◽

Streptococcus Pneumoniae ◽

Live Cell Imaging ◽

Cell Imaging ◽

Time Lapse ◽

Live Cell ◽

Time Lapse Microscopy

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Time-lapse live cell imaging to monitor doxorubicin release from DNA origami nanostructures

Journal of Materials Chemistry B ◽

10.1039/c7tb03223d ◽

2018 ◽

Vol 6 (11) ◽

pp. 1605-1612 ◽

Cited By ~ 19

Author(s):

Yun Zeng ◽

Jiajun Liu ◽

Shuo Yang ◽

Wenyan Liu ◽

Liang Xu ◽

...

Keyword(s):

Drug Delivery ◽

Live Cell Imaging ◽

Cell Imaging ◽

Time Lapse ◽

Dna Origami ◽

Live Cell ◽

Doxorubicin Release

DNA origami nanostructures can serve as a promising carrier for drug delivery due to the outstanding programmability and biocompatibility.

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Live cell imaging of macrophage/bacterium interaction demonstrates cell lysis induced by Corynebacterium diphtheriae and Corynebacterium ulcerans

BMC Research Notes ◽

10.1186/s13104-019-4733-y ◽

2019 ◽

Vol 12 (1) ◽

Author(s):

Dulanthi Weerasekera ◽

Jonas Hahn ◽

Martin Herrmann ◽

Andreas Burkovski

Keyword(s):

Fluorescence Microscopy ◽

Live Cell Imaging ◽

Cell Imaging ◽

Time Lapse ◽

Corynebacterium Diphtheriae ◽

Live Cell ◽

Human Macrophage ◽

Data Description ◽

Corynebacterium Ulcerans ◽

Microscopy Data

Abstract Objectives In frame of a study to characterize the interaction of human macrophage-like cells with pathogenic corynebacteria, Corynebacterium diphtheriae and Corynebacterium ulcerans, live cell imaging experiments were carried out and time lapse fluorescence microscopy videos were generated, which are presented here. Data description The time lapse fluorescence microscopy data revealed new insights in the interaction of corynebacteria with human macrophage-like THP-1 cells. In contrast to uninfected cells and infections with non-pathogenic C. glutamicum used as a control, pathogenic C. diphtheriae and C. ulcerans showed highly detrimental effects towards human cells and induction of cell death of macrophages.

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