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Cell Lysis Based on an Oscillating Microbubble Array.

ABSTRACT: Cell lysis is a process of breaking cell membranes to release intracellular substances such as DNA, RNA, protein, or organelles from a cell. The detection of DNA, RNA, or protein from the lysed cells is of importance for cancer diagnostics and drug screening. In this study, we develop a microbubble array that enables the realization of multiple cell lysis induced by the shear stress resulting from the individual oscillating microbubbles. The oscillating microbubbles in the channel have similar vibration amplitudes, and the intracellular substances can be released from the individual cells efficiently. Moreover, the efficiency of cell lysis increases with increments of input voltage and sonication time. By means of DNA agarose-gel electrophoresis, a sufficient extraction amount of DNA released from the lysed cells can be detected, and there is no significant difference in lysis efficiency when compared to cell lysis achieved using commercial kits. With the advantages of the simple manufacturing process, low cost, high efficiency, and high speed, this device can serve as an efficient and versatile tool for the single-cell sequencing of cell biology research, disease diagnosis, and stem cell therapy.

SUBMITTER: Liu X

PROVIDER: S-EPMC7143388 | biostudies-literature | 2020 Mar

REPOSITORIES: biostudies-literature

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Cell Lysis Based on an Oscillating Microbubble Array.

Liu Xiufang X Li Jinyuan J Zhang Liangyu L Huang Xiaowei X Farooq Umar U Pang Na N Zhou Wei W Qi Lin L Xu Lisheng L Niu Lili L Meng Long L

Micromachines 20200310 3

Cell lysis is a process of breaking cell membranes to release intracellular substances such as DNA, RNA, protein, or organelles from a cell. The detection of DNA, RNA, or protein from the lysed cells is of importance for cancer diagnostics and drug screening. In this study, we develop a microbubble array that enables the realization of multiple cell lysis induced by the shear stress resulting from the individual oscillating microbubbles. The oscillating microbubbles in the channel have similar v ...[more]

PMID: 32164279

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Project description:In this work, we discuss the development of H.O.S.T., a novel hemoglobin microbubble-based electrochemical biosensor for label-free detection of Hydrogen peroxide (H2O2) towards oxidative stress and cancer diagnostic applications. The novelty of the constructed sensor lies in the use of a sonochemically prepared hemoglobin microbubble capture probe, which allowed for an extended dynamic range, lower detection limit, and enhanced resolution compared to the native hemoglobin based H2O2 biosensors. The size of the prepared particles Hemoglobin microbubbles was characterized using Coulter Counter analysis and was found to be 4.4 microns, and the morphology of these spherical microbubbles was shown using Brightfield microscopy. The binding chemistry of the sensor stack elements of HbMbs' and P.A.N.H.S. crosslinker was characterized using Attenuated Total Reflectance Fourier Transform Infrared Spectroscopy and UV-Vis Spectroscopy. The electrochemical biosensor calibration (R2 > 0.95) was done using Electrochemical Impedance Spectroscopy, Cyclic Voltammetry, and Square Wave Voltammetry. The electrochemical biosensor calibration (R2 > 0.95) was done using Electrochemical Impedance Spectroscopy, Cyclic Voltammetry, and Square Wave Voltammetry. The specificity of the sensor for H2O2 was analyzed using cross-reactivity studies using ascorbic acid and glucose as interferents (p < 0.0001 for the highest non-specific dose versus the lowest specific dose). The developed sensor showed good agreement in performance with a commercially available kit for H2O2 detection using Bland Altman Analysis (mean bias = 0.37 for E.I.S. and - 24.26 for CV). The diagnostic potential of the biosensor was further tested in cancerous (N.G.P.) and non-cancerous (H.E.K.) cell lysate for H2O2 detection (p = 0.0064 for E.I.S. and p = 0.0062 for CV). The Michaelis Menten constant calculated from the linear portion of the sensor was found to be [Formula: see text] of 19.44 µM indicating that our biosensor has a higher affinity to Hydrogen peroxide than other available enzymatic sensors, it is attributed to the unique design of the hemoglobin polymers in microbubble.

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Cell Lysis Based on an Oscillating Microbubble Array.

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Cell Lysis Based on an Oscillating Microbubble Array.

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