Investigating dielectric properties of different stages of syngeneic murine ovarian cancer cells

Salmanzadeh, Alireza; Sano, Michael B.; Gallo‐Villanueva, Roberto C.; Roberts, Paul; Schmelz, Eva M.; Davalos, Rafael V.

doi:10.1063/1.4788921

Cited by 71 publications

(68 citation statements)

References 49 publications

(72 reference statements)

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“…The crossover frequency of MOSE-L FFLv cells is 11.9 kHz. 55 At 30 kHz the theoretical model with single shell sphere (Eqs. (1)- (3)) shows that the value of the Clausius-Mossotti factor is 0.5.…”

Section: Resultsmentioning

confidence: 99%

Enhanced contactless dielectrophoresis enrichment and isolation platform via cell-scale microstructures

et al. 2016

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We designed a new microfluidic device that uses pillars on the same order as the diameter of a cell (20 lm) to isolate and enrich rare cell samples from background. These cell-scale microstructures improve viability, trapping efficiency, and throughput while reducing pearl chaining. The area where cells trap on each pillar is small, such that only one or two cells trap while fluid flow carries away excess cells. We employed contactless dielectrophoresis in which a thin PDMS membrane separates the cell suspension from the electrodes, improving cell viability for off-chip collection and analysis. We compared viability and trapping efficiency of a highly aggressive Mouse Ovarian Surface Epithelial (MOSE) cell line in this 20 lm pillar device to measurements in an earlier device with the same layout but pillars of 100 lm diameter. We found that MOSE cells in the new device with 20 lm pillars had higher viability at 350 V RMS , 30 kHz, and 1.2 ml/h (control 77%, untrapped 71%, trapped 81%) than in the previous generation device (untrapped 47%, trapped 42%). The new device can trap up to 6 times more cells under the same conditions. Our new device can sort cells with a high flow rate of 2.2 ml/h and throughput of a few million cells per hour while maintaining a viable population of cells for off-chip analysis. By using the device to separate subpopulations of tumor cells while maintaining their viability at large sample sizes, this technology can be used in developing personalized treatments that target the most aggressive cancerous cells. V C 2016 AIP Publishing LLC. [http://dx

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Section: Resultsmentioning

confidence: 99%

Enhanced contactless dielectrophoresis enrichment and isolation platform via cell-scale microstructures

et al. 2016

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“…According to Wang et al, 41 Shim et al, 8 and Gascoyne et al, 42 this increase in surface area would putatively change the ruffles and folds in the cell membrane, increasing their effective surface area and specific membrane capacitance. Furthermore, Salmanzadeh et al 49 and Mulhall et al 50 observed that aggressiveness of cancer cell phenotype correlates with an increase in specific membrane capacitance for mouse ovarian surface epithelial and oral cancer cells, respectively. We, however, observed no significant change in the electrical properties of EMT-induced PANC1 cells.…”

Section: Resultsmentioning

confidence: 99%

Automated electrorotation shows electrokinetic separation of pancreatic cancer cells is robust to acquired chemotherapy resistance, serum starvation, and EMT

Lannin

Gruber

et al. 2016

Biomicrofluidics

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We used automated electrorotation to measure the cytoplasmic permittivity, cytoplasmic conductivity, and specific membrane capacitance of pancreatic cancer cells under environmental perturbation to evaluate the effects of serum starvation, epithelial-to-mesenchymal transition, and evolution of chemotherapy resistance which may be associated with the development and dissemination of cancer. First, we compared gemcitabine-resistant BxPC3 subclones with gemcitabine-naive parental cells. Second, we serum-starved BxPC3 and PANC-1 cells and compared them to untreated counterparts. Third, we induced the epithelial-to-mesenchymal transition in PANC-1 cells and compared them to untreated PANC-1 cells. We also measured the electrorotation spectra of white blood cells isolated from a healthy donor. The properties from fit electrorotation spectra were used to compute dielectrophoresis (DEP) spectra and crossover frequencies. For all three experiments, the median crossover frequency for both treated and untreated pancreatic cancer cells remained significantly lower than the median crossover frequency for white blood cells. The robustness of the crossover frequency to these treatments indicates that DEP is a promising technique for enhancing capture of circulating cancer cells. Published by AIP Publishing. [http://dx

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“…MOSE-L cells had a diameter of 17.7±3.3 μm (n = 268 cells) and the beads have a 4 μm nominal diameter. A full analysis of the dielectric properties of MOSE-L cells compared to MOSE cells in earlier stages of progression can be found in the recent work by Salmanzadeh et al 33 If pDEP and nDEP are not observed at these extremes, various problems could be occurring. The sample conductivity may be too high due to contaminants or cell lysing.…”

Section: Representative Resultsmentioning

confidence: 99%

“…To sort MOSE-L cells and 4 μm beads, operate the microdevice above 11.90±0.63 kHz. the first crossover frequency of MOSE-L cells recently reported by Salmanzadeh et al 33 3. To collect the contents of a sorted sample, remove the outlet tubing upon collecting the target volume, by placing a glove finger over the outlet opening and gently tugging on the tubing.…”

Section: Varying the Experiments To Perform Cell Sorting Or Enrichmentmentioning

confidence: 99%

“…[28][29][30][31] Lower frequency (5-100 kHz) devices operate continuously, and when operated at a frequency at which one population experiences pDEP while the background population experiences nDEP, can redirect particle trajectories to achieve sorting. [32][33][34] These low frequency devices have been used to sort cancer cells from red blood cells, determine the changes in dielectric properties of a progressive MOSE cell line, and to elucidate the effects of non-toxic sphingolipid treatments on reversing aggressive characteristics of aggressive MOSE cells. Additionally, cDEP microdevices can be designed to operate at increased throughput, currently up to 1 ml/hr.…”

Section: Introductionmentioning

confidence: 99%

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Label-free Isolation and Enrichment of Cells Through Contactless Dielectrophoresis

Elvington

Salmanzadeh

Stremler

et al. 2013

JoVE

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Dielectrophoresis (DEP) is the phenomenon by which polarized particles in a non-uniform electric field undergo translational motion, and can be used to direct the motion of microparticles in a surface marker-independent manner. Traditionally, DEP devices include planar metallic electrodes patterned in the sample channel. This approach can be expensive and requires a specialized cleanroom environment. Recently, a contact-free approach called contactless dielectrophoresis (cDEP) has been developed. This method utilizes the classic principle of DEP while avoiding direct contact between electrodes and sample by patterning fluidic electrodes and a sample channel from a single polydimethylsiloxane (PDMS) substrate, and has application as a rapid microfluidic strategy designed to sort and enrich microparticles. Unique to this method is that the electric field is generated via fluidic electrode channels containing a highly conductive fluid, which are separated from the sample channel by a thin insulating barrier. Because metal electrodes do not directly contact the sample, electrolysis, electrode delamination, and sample contamination are avoided. Additionally, this enables an inexpensive and simple fabrication process. cDEP is thus well-suited for manipulating sensitive biological particles. The dielectrophoretic force acting upon the particles depends not only upon spatial gradients of the electric field generated by customizable design of the device geometry, but the intrinsic biophysical properties of the cell. As such, cDEP is a label-free technique that avoids depending upon surface-expressed molecular biomarkers that may be variably expressed within a population, while still allowing characterization, enrichment, and sorting of bioparticles.Here, we demonstrate the basics of fabrication and experimentation using cDEP. We explain the simple preparation of a cDEP chip using soft lithography techniques. We discuss the experimental procedure for characterizing crossover frequency of a particle or cell, the frequency at which the dielectrophoretic force is zero. Finally, we demonstrate the use of this technique for sorting a mixture of ovarian cancer cells and fluorescing microspheres (beads).

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Investigating dielectric properties of different stages of syngeneic murine ovarian cancer cells

Cited by 71 publications

References 49 publications

Enhanced contactless dielectrophoresis enrichment and isolation platform via cell-scale microstructures

Enhanced contactless dielectrophoresis enrichment and isolation platform via cell-scale microstructures

Automated electrorotation shows electrokinetic separation of pancreatic cancer cells is robust to acquired chemotherapy resistance, serum starvation, and EMT

Label-free Isolation and Enrichment of Cells Through Contactless Dielectrophoresis

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