Label-free hybridoma cell culture quality control by a chip-based impedance flow cytometer

Pierzchalski, Arkadiusz; Hebeisen, Monika; Mittag, Anja; Bocsi, József; Berardino, Marco Di; Tárnok, Attila

doi:10.1039/c2lc40408g

Cited by 13 publications

(15 citation statements)

References 72 publications

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“…At higher scan frequencies the IFC discerns metabolic status [11]–[14], [16]. At a scan frequency of 9 MHz the IFC detects two populations of cells, the right-most population (higher phase values) reflects cells experiencing the initial stages of metabolic stress [11]-[14], [16]–[17].…”

Section: Resultsmentioning

confidence: 99%

“…At a scan frequency of 9 MHz the IFC detects two populations of cells, the right-most population (higher phase values) reflects cells experiencing the initial stages of metabolic stress [11]-[14], [16]–[17]. After three days of exposing MCF7 cells to PEMFs the magnitude of right-most population augmented, the greatest right-shift coinciding exactly with those parameters (3 mT, 20 Hz, for 60 min/day for 3 days) producing the greatest cell death in response to PEMFs (Fig 7 A-D).…”

Section: Resultsmentioning

confidence: 99%

“…For each measurement, 20,000 cells were analyzed at a frequency of 0.5 MHz to monitor apoptosis [11]–[13], [15] or 9 MHz to determine metabolic status [11]–[14], [16]–[17]. Each sample was run in parallel with polystyrene beads (8 µm) (Sigma-Aldrich, St. Louis, MO, USA) to obtain a standard signal response over the entire frequency spectrum, establishing a set point.…”

Section: Methodsmentioning

confidence: 99%

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Low Intensity and Frequency Pulsed Electromagnetic Fields Selectively Impair Breast Cancer Cell Viability

Crocetti

Beyer

Schade³

et al. 2013

PLoS ONE

110

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IntroductionA common drawback of many anticancer therapies is non-specificity in action of killing. We investigated the potential of ultra-low intensity and frequency pulsed electromagnetic fields (PEMFs) to kill breast cancer cells. Our criteria to accept this technology as a potentially valid therapeutic approach were: 1) cytotoxicity to breast cancer cells and; 2) that the designed fields proved innocuous to healthy cell classes that would be exposed to the PEMFs during clinical treatment.MethodsMCF7 breast cancer cells and their normal counterparts, MCF10 cells, were exposed to PEMFs and cytotoxic indices measured in order to design PEMF paradigms that best kill breast cancer cells. The PEMF parameters tested were: 1) frequencies ranging from 20 to 50 Hz; 2) intensities ranging from 2 mT to 5 mT and; 3) exposure durations ranging from 30 to 90 minutes per day for up to three days to determine the optimum parameters for selective cancer cell killing.ResultsWe observed a discrete window of vulnerability of MCF7 cells to PEMFs of 20 Hz frequency, 3 mT magnitude and exposure duration of 60 minutes per day. The cell damage accrued in response to PEMFs increased with time and gained significance after three days of consecutive daily exposure. By contrast, the PEMFs parameters determined to be most cytotoxic to breast cancer MCF-7 cells were not damaging to normal MCF-10 cells.ConclusionBased on our data it appears that PEMF-based anticancer strategies may represent a new therapeutic approach to treat breast cancer without affecting normal tissues in a manner that is non-invasive and can be potentially combined with existing anti-cancer treatments.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Low Intensity and Frequency Pulsed Electromagnetic Fields Selectively Impair Breast Cancer Cell Viability

Crocetti

Beyer

Schade³

et al. 2013

PLoS ONE

110

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“…IFC has been successfully used for cellular characterisation in bacterial and mammalian systems [6,8,9]. Analysing plant cells and in particular the quality of pollen in an efficient and reliable manner is not only of interest of plant breeding and seed production industries, but plays also a role in scientific projects investigating pollen development [30,50], pollen-related heat or cold tolerance [22,23], male-sterility systems [51], the establishment of pollen storage or doubled haploid protocols [13,14].…”

Section: Discussionmentioning

confidence: 99%

“…The analysis of electric properties in single cells by impedance flow cytometry (IFC) has been described in microorganisms to estimate viability, membrane potential, as well as cell size [6,7], in mammalian cells to characterize culture conditions and apoptosis [8] and to screen for methods to kill breast cancer cells [9]. The IFC system is based on the Coulter system [10] but uses a microfluidic chip which permits measurements in the radio frequency range from 0.1 to 30 MHz with alternating current (AC).…”

Section: Introductionmentioning

confidence: 99%

Impedance Flow Cytometry: A Novel Technique in Pollen Analysis

Heidmann

Schade-Kampmann²,

Lambalk

et al. 2016

PLoS ONE

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IntroductionAn efficient and reliable method to estimate plant cell viability, especially of pollen, is important for plant breeding research and plant production processes. Pollen quality is determined by classical methods, like staining techniques or in vitro pollen germination, each having disadvantages with respect to reliability, analysis speed, and species dependency. Analysing single cells based on their dielectric properties by impedance flow cytometry (IFC) has developed into a common method for cellular characterisation in microbiology and medicine during the last decade. The aim of this study is to demonstrate the potential of IFC in plant cell analysis with the focus on pollen.MethodDeveloping and mature pollen grains were analysed during their passage through a microfluidic chip to which radio frequencies of 0.5 to 12 MHz were applied. The acquired data provided information about the developmental stage, viability, and germination capacity. The biological relevance of the acquired IFC data was confirmed by classical staining methods, inactivation controls, as well as pollen germination assays.ResultsDifferent stages of developing pollen, dead, viable and germinating pollen populations could be detected and quantified by IFC. Pollen viability analysis by classical FDA staining showed a high correlation with IFC data. In parallel, pollen with active germination potential could be discriminated from the dead and the viable but non-germinating population.ConclusionThe presented data demonstrate that IFC is an efficient, label-free, reliable and non-destructive technique to analyse pollen quality in a species-independent manner.

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Impedance flow cytometry gauges proliferative capacity by detecting TRPC1 expression

et al. 2014

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When examined, the expansion of many stem cell classes has been shown to be facilitated by mechanically-regulated calcium entry from the extracellular space that also helps direct their developmental programs towards mechanosensitive tissues such as muscle, bone, and connective tissues. Cation channels of the transient receptor potential C class (TRPC) are the predominant conduit for calcium entry into proliferating myoblasts. Nonetheless, methods to non-invasively study this calcium-entry pathway are still in their infancy. Here we show that a microfluidic configuration of impedancebased flow cytometry (IFC) provides a method to detect TRP channel expression in cells at high throughput. Using this technology we discern changes in the IFC signal that correlates with the functional expression of TRPC1 channels and coincides with cell proliferation. Pharmacological agents, mechanical conditions or malignant states that alter the expression of TRPC1 channels are reflected in the IFC signal accordingly, whereas pharmacological agents that alter cation-permeation through TRPC1 channels, or ionophores that independently increase calcium entry across the membrane, have little effect. Our results suggest that IFC detects changes in whole-cell membrane organization associated with TRPC1 activation and surface expression, rather than cation permeation through the channel per se. IFC-based technologies thus have the potential to identify living stem cells in their earliest stages of expansion without staining or chemical fixation. V C 2014 International Society for Advancement of Cytometry

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Label-free hybridoma cell culture quality control by a chip-based impedance flow cytometer

Cited by 13 publications

References 72 publications

Low Intensity and Frequency Pulsed Electromagnetic Fields Selectively Impair Breast Cancer Cell Viability

Low Intensity and Frequency Pulsed Electromagnetic Fields Selectively Impair Breast Cancer Cell Viability

Impedance Flow Cytometry: A Novel Technique in Pollen Analysis

Impedance flow cytometry gauges proliferative capacity by detecting TRPC1 expression

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