Dual-frequency impedance assays for intracellular components in microalgal cells

Tang, Tao; Liu, Xun; Yuan, Yapeng; Kiya, Ryota; Shen, Yingjia; Zhang, Tianlong; Suzuki, Kengo; Li, Ming; Tanaka, Yo; Hosokawa, Yoichiroh; Yalikun, Yaxiaer

doi:10.1039/d1lc00721a

Cited by 20 publications

(36 citation statements)

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“…In this work, a frequency of 7 MHz is sufficient to penetrate the cell membrane for intracellular component detection. Our previous findings also support this conclusion 18,19 .…”

Section: Electrical Penetrationsupporting

confidence: 87%

“…Herein, E. gracilis cells were simulated as single-shell ellipses (radius: 30 μm long-axis, 10 μm short-axis), and the cell membrane (10 nm) was modeled using contact impedance approximation 23 . Intracellular components were simplified as 2D circles (membrane thickness of 20 nm and diameter of 1 μm) and were closely placed in the left interior of the cell 18,19 , as shown in Fig. 2a.…”

Section: Tilt Indexmentioning

confidence: 99%

“…2a. Other parameters of cells 18 used in the simulation are listed in Table S1 in the Supplementary information.…”

Section: Tilt Indexmentioning

confidence: 99%

“…Our solution is to quantify the tilt level of impedance pulses as a tilt index 13 and then assess the conductivity of the cell membrane through the tilt index of the cell population at different detection frequencies. Based on our previous work, the intracellular component distribution was found to affect the tilting level of high-frequency impedance pulses (6 MHz) 18,19 because a high-frequency current can propagate inside single cells between nonconductive intracellular components. In contrast, a low-frequency current (500 kHz) cannot penetrate the cell membrane, and it mainly propagates around the cell 18,19 .…”

Section: Introductionmentioning

confidence: 99%

“…Based on our previous work, the intracellular component distribution was found to affect the tilting level of high-frequency impedance pulses (6 MHz) 18,19 because a high-frequency current can propagate inside single cells between nonconductive intracellular components. In contrast, a low-frequency current (500 kHz) cannot penetrate the cell membrane, and it mainly propagates around the cell 18,19 . This feature facilitates a novel method for determining the detection frequency of the cell interior and exterior based on the tilt index of impedance pulses.…”

Section: Introductionmentioning

confidence: 99%

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Assessment of the electrical penetration of cell membranes using four-frequency impedance cytometry

et al. 2022

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The electrical penetration of the cell membrane is vital for determining the cell interior via impedance cytometry. Herein, we propose a method for determining the conductivity of the cell membrane through the tilting levels of impedance pulses. When electrical penetration occurs, a high-frequency current freely passes through the cell membrane; thus, the intracellular distribution can directly act on the high-frequency impedance pulses. Numerical simulation shows that an uneven intracellular component distribution can affect the tilting levels of impedance pulses, and the tilting levels start increasing when the cell membrane is electrically penetrated. Experimental evidence shows that higher detection frequencies (>7 MHz) lead to a wider distribution of the tilting levels of impedance pulses when measuring cell populations with four-frequency impedance cytometry. This finding allows us to determine that a detection frequency of 7 MHz is able to pass through the membrane of Euglena gracilis (E. gracilis) cells. Additionally, we provide a possible application of four-frequency impedance cytometry in the biomass monitoring of single E. gracilis cells. High-frequency impedance (≥7 MHz) can be applied to monitor these biomass changes, and low-frequency impedance (<7 MHz) can be applied to track the corresponding biovolume changes. Overall, this work demonstrates an easy determination method for the electrical penetration of the cell membrane, and the proposed platform is applicable for the multiparameter assessment of the cell state during cultivation.

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“…In this work, a frequency of 7 MHz is sufficient to penetrate the cell membrane for intracellular component detection. Our previous findings also support this conclusion 18,19 .…”

Section: Electrical Penetrationsupporting

confidence: 87%

Section: Tilt Indexmentioning

confidence: 99%

“…2a. Other parameters of cells 18 used in the simulation are listed in Table S1 in the Supplementary information.…”

Section: Tilt Indexmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Assessment of the electrical penetration of cell membranes using four-frequency impedance cytometry

et al. 2022

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Microfluidic characterization of single‐cell biophysical properties and the applications in cancer diagnosis

Li,

Xue,

et al. 2023

Electrophoresis

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Single‐cell biophysical properties play a crucial role in regulating cellular physiological states and functions, demonstrating significant potential in the fields of life sciences and clinical diagnostics. Therefore, over the last few decades, researchers have developed various detection tools to explore the relationship between the biophysical changes of biological cells and human diseases. With the rapid advancement of modern microfabrication technology, microfluidic devices have quickly emerged as a promising platform for single‐cell analysis offering advantages including high‐throughput, exceptional precision, and ease of manipulation. Consequently, this paper provides an overview of the recent advances in microfluidic analysis and detection systems for single‐cell biophysical properties and their applications in the field of cancer. The working principles and latest research progress of single‐cell biophysical property detection are first analyzed, highlighting the significance of electrical and mechanical properties. The development of data acquisition and processing methods for real‐time, high‐throughput, and practical applications are then discussed. Furthermore, the differences in biophysical properties between tumor and normal cells are outlined, illustrating the potential for utilizing single‐cell biophysical properties for tumor cell identification, classification, and drug response assessment. Lastly, we summarize the limitations of existing microfluidic analysis and detection systems in single‐cell biophysical properties, while also pointing out the prospects and future directions of their applications in cancer diagnosis and treatment.

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Impedance‐Enabled Camera‐Free Intrinsic Mechanical Cytometry

Feng

Chai

et al. 2022

Small Methods

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and isotopes), requiring tedious sample preparation, invasive operation, and prior knowledge of cell specificity. [3] In contrast, biophysical methods measure the mechanical, electrical, and optical properties to perform quick, label-free, and non-invasive single-cell characterization, [4] so as to facilitate subsequent operations on cells such as sorting, [5] culturing, [6] and omics analysis. [7] Cellular biophysical properties have been proven as effective biomarkers for diagnosing diseases and widely used in cell biology, dynamics, and pathology research. [8] As one of the biophysical properties, mechanical properties of cells reflect the characteristics of cell membranes, and cytoskeletal networks, which influence cellular and subcellular functions, [9] including cell adhesion, migration, polarization, and differentiation, as well as organelle organization and trafficking inside the cytoplasm. [10] At present, there are many classic methods for characterizing the mechanical properties of single cells, such as atomic force microscopy (AFM), micropipette aspiration, parallel-plate rheometry, and optical stretching. [11] These methods evaluate time-resolved cellular responses to external stresses, and offer effective means to obtain the single-cell intrinsic mechanical properties (e.g., Young's modulus and viscosity). However, they suffer from inherent problems with technically demanding and time-consuming procedures, which lead to difficulty in operation and low throughput (<1 cell min −1 ). [11] As an attractive alternative, microfluidics approaches allow detecting cell deformability at high throughput but low complexity for single-cell analysis. [12] However, the characterization relies on expensive and complex high-speed imaging systems with time-consuming image processing algorithms to obtain the dynamic deformation process of the cells, which limits the real-time capability desired for downstream cell manipulation and analysis such as deformability-based sorting.Exploiting electronic means or impedance cytometry for cellular mechanical characterization presents a new alternative approach. [13] Based on direct-current (DC) Coulter-counter, the visco-node-pore sensing system [14] is proposed to characterize the cellular mechanics solely by the current signal. It employs elegant design of straight [14] or sinusoidal [15] constriction channels to correlate the current signal to the mechanical viscoelastic properties, and is applied in biological applications. [16] But the footprint for the series of node-pore structure may limit Mechanical properties of single cells are important label-free biomarkers normally measured by expensive and complex imaging systems. To unlock this limit and allow mechanical properties comparable across different measurement platforms, camera-free intrinsic mechanical cytometry (CFIMC) is proposed for on-the-fly measurement of two major intrinsic mechanical parameters, that is, Young's modulus E and fluidity β, of single cells. CFIMC adopts a framework that couples the impedance ele...

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Dual-frequency impedance assays for intracellular components in microalgal cells

Abstract: Intracellular components (including organelles and biomolecules) at the submicron level are typically analyzed in situ by special preparation or expensive setups. Here, a label-free and cost-effective approach of screening microalgal...

Cited by 20 publications

References 49 publications

Assessment of the electrical penetration of cell membranes using four-frequency impedance cytometry

Assessment of the electrical penetration of cell membranes using four-frequency impedance cytometry

Microfluidic characterization of single‐cell biophysical properties and the applications in cancer diagnosis

Impedance‐Enabled Camera‐Free Intrinsic Mechanical Cytometry

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