Microscopic impedance cytometry for quantifying single cell shape

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

doi:10.1016/j.bios.2021.113521

Cited by 27 publications

(24 citation statements)

References 43 publications

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“…The detection mechanism is supported by numerous previous works. Our previous work has shown that low-frequency detection is applicable for analyzing cell morphology, as the low-frequency current mainly propagates around the cell 13,18,19 . The dependence of low-frequency impedance on cell volume has also been verified 8 .…”

Section: Discussionmentioning

confidence: 99%

“…Each electrode is coated with a 70 nm thick layer of gold (Au) over a 70 nm thick layer of chromium (Cr). The complete fabrication procedure was described in detail in our previous work 13,18 .…”

Section: Impedance Microfluidic Devicesmentioning

confidence: 99%

“…As shown in Fig. 1c, a field-programmable gate array (FPGA)-based lock-in amplifier (Diligent Eclypse Z7, USA) was used to generate a 1 V alternative current (AC) signal at four detection frequencies (i.e., 500 kHz, 4 MHz, 7 MHz, and 10 MHz) and perform the real-time processing of the impedance signals 13,20 from the detection area. The current signals from two detection electrodes were converted into voltage signals with self-developed transimpedance amplifiers (I/V converters) and then compared with differential amplifiers (Diff).…”

Section: Impedance Detection and Calibrationmentioning

confidence: 99%

“…To date, impedance cytometry has been successfully employed to analyze the morphology 9 , stiffness 10 , and states 11 of single cells. The magnitude and morphology of impedance pulses have been shown to be dependent on the volume 12 and shape 13 of single cells, respectively. In addition, research has found that high-frequency impedance detection is applicable for characterizing membrane properties 14,15 .…”

Section: Introductionmentioning

confidence: 99%

“…When measuring intracellular biomass with impedance cytometry, it is necessary to determine the detection frequency that can penetrate the cell membrane. 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.…”

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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Section: Discussionmentioning

confidence: 99%

Section: Impedance Microfluidic Devicesmentioning

confidence: 99%

Section: Impedance Detection and Calibrationmentioning

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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Robust Spontaneous Raman Flow Cytometry for Single‐Cell Metabolic Phenome Profiling via pDEP‐DLD‐RFC

et al. 2023

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A full-spectrum spontaneous single-cell Raman spectrum (fs-SCRS) captures the metabolic phenome for a given cellular state of the cell in a label-free, landscape-like manner. Herein a positive dielectrophoresis induced deterministic lateral displacement-based Raman flow cytometry (pDEP-DLD-RFC) is established. This robust flow cytometry platform utilizes a periodical positive dielectrophoresis induced deterministic lateral displacement (pDEP-DLD) force that is exerted to focus and trap fast-moving single cells in a wide channel, which enables efficient fs-SCRS acquisition and extended stable running time. It automatically produces deeply sampled, heterogeneity-resolved, and highly reproducible ramanomes for isogenic cell populations of yeast, microalgae, bacteria, and human cancers, which support biosynthetic process dissection, antimicrobial susceptibility profiling, and cell-type classification. Moreover, when coupled with intra-ramanome correlation analysis, it reveals state-and cell-type-specific metabolic heterogeneity and metabolite-conversion networks. The throughput of ≈30-2700 events min −1 for profiling both nonresonance and resonance marker bands in a fs-SCRS, plus the >5 h stable running time, represent the highest performance among reported spontaneous Raman flow cytometry (RFC) systems. Therefore, pDEP-DLD-RFC is a valuable new tool for label-free, noninvasive, and high-throughput profiling of single-cell metabolic phenomes.

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Electrical micro flow cytometry with LSTM and its application in leukocyte differential

Tan,

Chen,

Huang

et al. 2023

Cytometry Pt A

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This paper developed an electrical micro flow cytometry to realize leukocyte differentials leveraging a constrictional microchannel and a deep neural network. Firstly, purified granulocytes, lymphocytes or monocytes traveled through the constrictional microchannel with a cross‐sectional area marginally larger than individual cells and produced large impedance variations by blocking focused electric field lines. By optimizing key elements (e.g., normalization, learning rate, batch size and neuron number) of the recurrent neural network (RNN), electrical results of purified leukocytes were analyzed to establish a leukocyte differential system with a classification accuracy of 95.2%. Then the leukocyte mixtures were forced to travel through the same constrictional microchannel, producing mixed impedance profiles which were classified into granulocytes, lymphocytes and monocytes based on the aforementioned differential system. As to the classification results, two leukocyte mixtures from the same donor were processed, producing comparable classification results, which were 57% versus 59% of granulocytes, 37% versus 34% of lymphocytes and 6% versus 7% of monocytes. These results validated the established classification system based on the constrictional microchannel and the recurrent neural network, providing a new perspective of differentiating white blood cells by electrical flow cytometry.

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Microscopic impedance cytometry for quantifying single cell shape

Cited by 27 publications

References 43 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

Robust Spontaneous Raman Flow Cytometry for Single‐Cell Metabolic Phenome Profiling via pDEP‐DLD‐RFC

Electrical micro flow cytometry with LSTM and its application in leukocyte differential

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