Nanoscale tweezers for spatially resolved single-cell analysis

Quantitatively assessing the spatial distribution of biomolecules in individual cells can aid our understanding of the roles that they have within a cell and therefore help us elucidate their molecular behaviour under different cellular conditions and in disease. Subcellular extraction methods have the potential to achieve this via multi-omics analyses from single cells, without the need for lysing or fixing whole cells. However, current intracellular analysis tools are limited in their ability to perform repeated spatially controlled measurements of the same cell because they involve the aspiration of cytoplasmic fluid, cellular alterations, or the destruction of the cell. They also have costly and complicated fabrication procedures. The nanotweezer is a tool that circumvents these issues via direct trapping of biomolecules using dielectrophoresis, allowing for minimally invasive extraction of DNA, RNA and single organelles from living cells, whilst utilising a simple fabrication procedure. Here, we show that the nanotweezer can be used to extract nucleic acids and organelles from subcellular compartments of live single neurons with high spatial precision. Repeat sampling of the same neuron is achieved without affecting the cells’ viability, microenvironment, and neuronal activity. Further, we show that single organelle and mRNA analysis from individual biopsies can be achieved, highlighting the prominence of this technique for precise quantification and mapping of biomolecules in the same neuron.