研究目的
To develop minimally invasive nanotweezers for the spatial control and extraction of samples from living cells with single-molecule precision, enabling real-time analysis and perturbation of living cells.
研究成果
The developed nanotweezers enable minimally invasive extraction of intracellular molecules with single-molecule precision, offering a powerful tool for real-time analysis of living cells. This technology bridges the gap between single-molecule/organelle manipulation and cell biology, promising advancements in understanding cellular heterogeneity and dynamics.
研究不足
The study demonstrates the potential of nanotweezers for single-cell biopsies but notes the need for further optimization for broader application, including the handling of low copy number mRNAs and the minimization of potential cell damage during extraction.
1：Experimental Design and Method Selection:
The study utilized nanotweezers composed of two closely spaced electrodes for dielectrophoretic trapping of DNA and proteins. The methodology included the fabrication of nanotweezers, their characterization, and application in single-cell biopsies.
2：Sample Selection and Data Sources:
Human osteosarcoma (U2OS) cells and primary human pulmonary artery endothelial cells (HPAECs) were used for nucleic acid extraction. Primary rodent hippocampal neurons were used for mitochondrion extraction.
3：List of Experimental Equipment and Materials:
Double-barrelled quartz capillaries for nanotweezer fabrication, YOYO-1 for DNA labelling, SYTO RNASelect for RNA staining, and MitoTracker Green for mitochondrion visualization.
4：Experimental Procedures and Operational Workflow:
The nanotweezers were inserted into cells, and an a.c. voltage was applied for trapping. The extracted material was then analyzed using qPCR and sequencing.
5：Data Analysis Methods:
Fluorescence microscopy for visualization, qPCR for amplification, and sequencing for confirmation of extracted DNA integrity.
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