The electroporation system can serve as a tool for the intracellular delivery of foreign cargos. However, this technique is presently limited by the inaccurate electric field applied to the single cells and lack of a real-time electroporation metrics subsystem. Here, we reported a microfluidic system for precise and rapid single-cell electroporation and simultaneous impedance monitoring in a constriction microchannel. When single cells (A549) were continuously passing through the constriction microchannel, a localized high electric field was applied on the cell membrane, which resulted in highly efficient (up to 96.6%) electroporation. During a single cell entering the constriction channel, an abrupt impedance drop was noticed and demonstrated to be correlated with the occurrence of electroporation. Besides, while the cell was moving in the constriction channel, the stabilized impedance showed the capability to quantify the electroporation extent. The correspondence of the impedance variation and electroporation was validated by the intracellular delivery of the fluorescence indicator (propidium iodide). Based on the obtained results, this system is capable of precise control of electroporation and real-time, label-free impedance assessment, providing a potential tool for intracellular delivery and other biomedical applications.
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http://dx.doi.org/10.3390/mi11090856 | DOI Listing |
Proc Natl Acad Sci U S A
December 2024
Mechanobiology Institute, National University of Singapore, 117411, Singapore.
Diverse tissues in vivo present varying degrees of confinement, constriction, and compression to migrating cells in both homeostasis and disease. The nucleus in particular is subjected to external forces by the physical environment during confined migration. While many systems have been developed to induce nuclear deformation and analyze resultant functional changes, much remains unclear about dynamic volume regulation in confinement due to limitations in time resolution and difficulty imaging in PDMS-based microfluidic chips.
View Article and Find Full Text PDFMicrosyst Nanoeng
December 2024
State Key Laboratory of Transducer Technology, Aerospace Information Research Institute, Chinese Academy of Sciences, Beijing, 100190, People's Republic of China.
Microfluidic impedance flow cytometry has been widely used in leukocyte differential and counting, but it faces a bottleneck due to the trade-off between impedance detection throughput and sensitivity. In this study, a microfluidic impedance flow cytometer based on a virtual constriction microchannel was reported, in which the virtual constriction microchannel was constructed by crossflow of conductive sample and insulated sheath fluids with underneath micro-electrodes for impedance measurements. Compared to conventional mechanical constriction microchannels, this virtual counterpart could effectively avoid direct physical contact between cells and the microchannel walls to maintain high throughputs, and significantly reduce the volume of the impedance detection region for sensitivity improvements.
View Article and Find Full Text PDFLab Chip
November 2024
Laboratory of Biosensors and Bioelectronics (LBB), ETH Zürich, 8092 Zürich, Switzerland.
Technologies for axon guidance for disease models and bottom up investigations are increasingly being used in neuroscience research. One of the most prevalent patterning methods is using polydimethylsiloxane (PDMS) microstructures due to compatibility with microscopy and electrophysiology which enables systematic tracking of axon development with precision and efficiency. Previous investigations of these guidance platforms have noted axons tend to follow edges and avoid sharp turns; however, the specific impact of spatial constraints remains only partially explored.
View Article and Find Full Text PDFAnal Chem
November 2024
Institute of Oncology, Cancer Hospital of Dalian University of Technology, Shenyang, Liaoning 110042, P. R. China.
The microfluidic impedance flow cytometer (m-IFC) using constricted microchannels is an appealing choice for the high-throughput measurement of single-cell mechanical properties. However, channels smaller than the cells are susceptible to irreversible blockage, extremely affecting the stability of the system and the throughput. Meanwhile, the common practice of extracting a single quantitative index, i.
View Article and Find Full Text PDFBr J Haematol
December 2024
Dr. Senckenberg Institute of Pathology, Goethe University Frankfurt am Main, Frankfurt am Main, Germany.
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