Sensing of Yeast Inactivation by Electroporation

Pintarelli, Guilherme Brasil; Ramos, Corinne; Silva, J. R. da; Rossi, Márcio José; Suzuki, Daniela Ota Hisayasu

doi:10.1109/jsen.2021.3066092

Cited by 6 publications

(8 citation statements)

References 52 publications

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“…Because most of the dispersion is created only by the cell membrane, it tends to have the dispersion center at low frequency due to the thin membrane (the dispersion center, in this case, is near 10 kHz). The σHigh medium is more typical for liquid foods [4,38,46]. The σMed and σHigh buffers are more similar regarding dispersion center location.…”

Section: Discussionmentioning

confidence: 93%

“…COMSOL Multiphysics ® version 4.4 software is used for FEM modeling and 2D electric currents application ('ec', from the COMSOL's AC/DC Module). The electroporation model is solved in the time domain (as described previously [4,31]), giving results of pore density N(t) and average membrane electrical conductivity (see Appendix A Equations (A1) and (A2)). The simulation steps are defined as linear and were 0.5 µs when using 50 µs PEF and 0.01 µs when using 1 µs PEF.…”

Section: Numerical Modelingmentioning

confidence: 99%

“…Pulsed Electric Field (PEF) is a non-thermal electricity-based process to control cells. PEF is used in bio-technology [1], veterinary [2] and medical research [3], and food industry [4,5]. These applications require specific levels of electroporation control.…”

Section: Introductionmentioning

confidence: 99%

“…Thus, PEF systems usually do not have electroporation feedback because there is no adequate PEF probing method. We have recently shown that it is possible to eliminate yeast using irreversible electroporation and the macroscopic current changes due to cell breakage [4]. Yeasts are a problem in the industry, as their contamination can cause financial loss.…”

Section: Introductionmentioning

confidence: 99%

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Dielectric Dispersion Modulated Sensing of Yeast Suspension Electroporation

Pintarelli

Silva

Yang

et al. 2022

Sensors

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A specific pulsed electric field protocol can be used to induce electroporation. This is used in the food industry for yeast pasteurization, in laboratories for generic transfer and the medical field for cancer treatment. The sensing of electroporation can be done with simple ‘instantaneous’ voltage-current analysis. However, there are some intrinsic low-frequency phenomena superposing the electroporation current, such as electrode polarization. The biological media are non-homogeneous, giving them specific characterization in the broad frequency spectrum. For example, the cell barrier, i.e., cell membrane, causes so called β-dispersion in the frequency range of tens to thousands of kHz. Electroporation is a dynamic phenomenon characterized by altering the cell membrane permeability. In this work, we show that the impedance measurement at certain frequencies could be used to detect the occurrence of electroporation, i.e., dielectric dispersion modulated sensing. This approach may be used for the design and implementation of electroporation systems. Yeast suspension electroporation is simulated to show changes in the frequency spectrum. Moreover, the alteration depends on characteristics of the system. Three types of external buffers and their characteristics are evaluated.

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Section: Discussionmentioning

confidence: 93%

Section: Numerical Modelingmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Dielectric Dispersion Modulated Sensing of Yeast Suspension Electroporation

Pintarelli

Silva

Yang

et al. 2022

Sensors

Self Cite

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“…Among these methods, DS is a non-invasive, fast, and promising method for cell screening. DS has been widely used for impedance measurements of yeast cells [10][11][12], breast cancer cells [13], lymphocyte activation profiling [14], monitoring performance of epithelial tissues [15], and hepatology research [16].…”

Section: Introductionmentioning

confidence: 99%

A Microfluidic Dielectric Spectroscopy System for Characterization of Biological Cells in Physiological Media

Bakhtiari

Manshadi

Mansoorifar

et al. 2022

Sensors

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Dielectric spectroscopy (DS) is a promising cell screening method that can be used for diagnostic and drug discovery purposes. The primary challenge of using DS in physiological buffers is the electrode polarization (EP) that overwhelms the impedance signal within a large frequency range. These effects further amplify with the miniaturization of the measurement electrodes. In this study, we present a microfluidic system and the associated equivalent circuit models for real-time measurements of cell membrane capacitance and cytoplasm resistance in physiological buffers with 10 s increments. The current device captures several hundreds of biological cells in individual microwells through gravitational settling and measures the system’s impedance using microelectrodes covered with dendritic gold nanostructures. Using PC-3 cells (a highly metastatic prostate cancer cell line) suspended in cell growth media (CGM), we demonstrate stable measurements of cell membrane capacitance and cytoplasm resistance in the device for over 15 min. We also describe a consistent application of the equivalent circuit model, starting from the reference measurements used to determine the system parameters. The circuit model is tested using devices with varying dimensions, and the obtained cell parameters between different devices are nearly identical. Further analyses of the impedance data have shown that accurate cell membrane capacitance and cytoplasm resistance can be extracted using a limited number of measurements in the 5 MHz to 10 MHz range. This will potentially reduce the timescale required for real-time DS measurements below 1 s. Overall, the new microfluidic device can be used for the dielectric characterization of biological cells in physiological buffers for various cell screening applications.

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Metabolomics of astaxanthin biosynthesis and corresponding regulation strategies in Phaffia rhodozyma

Yang

et al. 2023

Yeast

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Astaxanthin is a valuable carotenoid and is used as antioxidant and health care. Phaffia rhodozyma is a potential strain for the biosynthesis of astaxanthin. The unclear metabolic characteristics of P. rhodozyma at different metabolic stages hinder astaxanthin's promotion. This study is conducted to investigate metabolite changes based on quadrupole time‐of‐flight mass spectrometry metabolomics method. The results showed that the downregulation of purine, pyrimidine, amino acid synthesis, and glycolytic pathways contributed to astaxanthin biosynthesis. Meanwhile, the upregulation of lipid metabolites contributed to astaxanthin accumulation. Therefore, the regulation strategies were proposed based on this. The addition of sodium orthovanadate inhibited the amino acid pathway to increase astaxanthin concentration by 19.2%. And the addition of melatonin promoted lipid metabolism to increase the astaxanthin concentration by 30.3%. It further confirmed that inhibition of amino acid metabolism and promotion of lipid metabolism were beneficial for astaxanthin biosynthesis of P. rhodozyma. It is helpful in understanding metabolic pathways affecting astaxanthin of P. rhodozyma and provides regulatory strategies for metabolism.

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Sensing of Yeast Inactivation by Electroporation

Cited by 6 publications

References 52 publications

Dielectric Dispersion Modulated Sensing of Yeast Suspension Electroporation

Dielectric Dispersion Modulated Sensing of Yeast Suspension Electroporation

A Microfluidic Dielectric Spectroscopy System for Characterization of Biological Cells in Physiological Media

Metabolomics of astaxanthin biosynthesis and corresponding regulation strategies in Phaffia rhodozyma

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