Single cell dielectrophoresis study of apoptosis progression induced by controlled starvation

Salimi, Elham; Braasch, Katrin; Fazelkhah, Azita; Afshar, Samaneh; Rizi, Bahareh Saboktakin; Mohammad, Kaveh; Butler, Michael; Bridges, Greg E.; Thomson, D. J.

doi:10.1016/j.bioelechem.2018.07.003

Cited by 13 publications

(10 citation statements)

References 39 publications

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“…The decrease in the FI occurred after 72–96 hr of the culture where the nutrients in the medium had been mostly consumed by the cells and the decline progressed quickly over the next 24 hr. This behavior is similar to nutrient deprivation results (Salimi et al, ). This suggests that the threshold of 0.3 S/m can be used as the minimum cytoplasm conductivity that allows restoration of the viability following nutrient reintroduction.…”

Section: Resultssupporting

confidence: 91%

“…If the nutrient deprivation is extended beyond 36 hr, cells become apoptotic and are no longer able to restore the cytoplasm conductivity to the normal viable level with the reintroduction of nutrients. A much greater loss in ions and hence decrease in cytoplasm conductivity occurs for apoptotic cells (Afshar et al, ; Hughes, Bortner, Purdy, & Cidlowski, ; Labeed, Coley, & Hughes, ; McCarthy & Cotter, ; Mulhall, Cardnell, Hoettges, Labeed, & Hughes, ; Salimi et al, ; Yu, Canzoniero, & Choi, ). At 48 hr when glucose is reintroduced to the medium, no significant change in the status of cells is observed and the increase in conductivity is minimal.…”

Section: Resultsmentioning

confidence: 99%

“…Electromagnetic models that are employed to extract the properties of the cells often neglect the impact of cell population heterogeneity and provide the average properties of all cells (Ansorge et al, ; Opel et al, ). In our previous work, we employed a single cell DEP technique to study the change in the cytoplasm conductivity of cells (Salimi et al, ). We showed that nutrient depletion in the culture was associated with a decrease in dielectric response of the cells and closely correlated with the cells’ cytoplasm conductivity (Afshar et al, ; Salimi et al, ).…”

Section: Introductionmentioning

confidence: 99%

“…In our previous work, we employed a single cell DEP technique to study the change in the cytoplasm conductivity of cells (Salimi et al, ). We showed that nutrient depletion in the culture was associated with a decrease in dielectric response of the cells and closely correlated with the cells’ cytoplasm conductivity (Afshar et al, ; Salimi et al, ).…”

Section: Introductionmentioning

confidence: 99%

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Cytoplasmic conductivity as a marker for bioprocess monitoring: Study of Chinese hamster ovary cells under nutrient deprivation and reintroduction

Fazelkhah

Afshar

Braasch

et al. 2019

Biotech & Bioengineering

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The ability to monitor the status of cells during nutrient limitation is important for optimizing bioprocess growth conditions in batch and fed‐batch cultures. The activity level of Na+/K+ ATPase pumps and cytoplasm ionic concentrations are directly influenced by the nutrient level, and thus, cytoplasm conductivity can be used as a markerless indicator of cell status. In this work, we monitored the change in cytoplasm conductivity of Chinese hamster ovary (CHO) cells during nutrient deprivation and reintroduction. Employing single cell dielectrophoresis, the change in cytoplasm conductivity was measured over a 48‐hr period. The conditions under which the cytoplasm conductivity would recover to a normal level after nutrient reintroduction was determined. In addition, numerical simulations of cell ion flux, for different levels of Na+/K+ ATPase pump inhibition, were used to predict the minimum conductivity expected for nutrient‐deprived CHO cells. This predicted value is close to the minimum observed experimental cytoplasm conductivity for CHO cells that maintain the ability to restore the cytoplasm conductivity to the normal viable levels when nutrients are reintroduced. The recovery of starved cells was verified by reintroducing them to nutrient for 36 hr and measuring their proliferation using trypan blue exclusion assay. We conclude that cytoplasm conductivity can be used as a marker to indicate whether cells are in a recoverable state, such that the reintroduction of nutrients results in cells returning to a normal healthy state.

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Section: Resultssupporting

confidence: 91%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Cytoplasmic conductivity as a marker for bioprocess monitoring: Study of Chinese hamster ovary cells under nutrient deprivation and reintroduction

Fazelkhah

Afshar

Braasch

et al. 2019

Biotech & Bioengineering

Self Cite

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“…Viability assessment of some mammalian cells like human B cells , human sperm , chondrocytes from steers , Chinese hamster ovary cells , human epithermal breast cells , and human stromal cells were also investigated with DEP.…”

Section: Cell Viability Assessmentmentioning

confidence: 99%

Recent advances in dielectrophoresis‐based cell viability assessment

et al. 2020

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Recent advances in dielectrophoresis-based cell viability assessmentDielectrophoresis (DEP) is a non-destructive, accurate, and label-free cell manipulating technique and DEP applications have been found in various fields. Assessment of cell viability is one of the important applications and many investigations have been reported. In this paper, cell polarization and its modeling, some key parameters employed for living/dead cell separation, as well as electrode configurations are reviewed. Focus is given to the latest development of DEP devices employed for the assessment of cell viability. Experimentally determined factors for separating living/dead cells, such as the conductivity of suspending medium and the frequency of applied electric field, are summarized. The future directions and potential challenges in this field are also outlined. Theory of cell DEPThe fundamentals of DEP were systematically reviewed in the past [35,36]. In this section, we only focus on the theory concerning cellular DEP. Based on the spherical particle model, single-shell model and multi-shell model of biological cells are extended. Color online: See the article online to view Figs. 1-7 in color. J. Zhang et al. Electrophoresis 2020, 41, 917-932

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Parallel single‐cell optical transit dielectrophoresis cytometer

et al. 2020

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In this work, we present an optical transit DEP flow cytometer for parallel single-cell analysis. Each cell's dielectric property is inferred from velocity perturbations due to DEP actuation in a microfluidic channel. Dual LED sources facilitate velocity measurement by producing two transit shadows for each cell passing through the channel. These shadows are detected using a 256-pixel linear optical array detector. Massively parallel analysis is possible as each pixel of the detector can independently analyze the passing cells. A wide channel (∼18 mm) was employed to carry many particles simultaneously, and the system was capable of detecting the velocity of over 200 cells simultaneously. We have achieved analysis rates for 10 µm diameter polystyrene spheres response exceeding 250 per second. With appropriate calibration, this DEP cytometer can quantitatively measure the dielectric response. The dielectric response (Clausius-Mossotti factor) of viable CHO cells was measured over the frequency range of 100 kHz to 6 MHz, and the obtained response matches the previously measured values by our group. The DEP cytometer uses simple modular components to achieve high throughput label-free single-cell dielectric analysis and can begin analyzing particles within 10 s after starting to pump the sample into the channel.

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Single cell dielectrophoresis study of apoptosis progression induced by controlled starvation

Cited by 13 publications

References 39 publications

Cytoplasmic conductivity as a marker for bioprocess monitoring: Study of Chinese hamster ovary cells under nutrient deprivation and reintroduction

Cytoplasmic conductivity as a marker for bioprocess monitoring: Study of Chinese hamster ovary cells under nutrient deprivation and reintroduction

Recent advances in dielectrophoresis‐based cell viability assessment

Parallel single‐cell optical transit dielectrophoresis cytometer

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