Physics and Astronomy, Faculty of Physical Sciences and Engineering, University of Southampton, Southampton, SO17 1BJ, UK.
Chemistry Research Laboratory, Department of Chemistry, University of Oxford, 12 Mansfield Road, Oxford, OX1 3TA, UK.
Small. 2018 Mar;14(12):e1703489. doi: 10.1002/smll.201703489. Epub 2018 Feb 21.
Wound healing is a highly complex biological process, which is accompanied by changes in cell phenotype, variations in protein expression, and the production of active biomolecules. Currently, the detection of proteins in cells is done by immunostaining where the proteins in fixed cells are detected by labeled antibodies. However, immunostaining cannot provide information about dynamic processes in living cells, within the whole tissue. Here, an easy method is presented to detect the transition of epithelial to mesenchymal cells during wound healing. The method employs DNA-coated gold nanoparticle fluorescent nanoprobes to sense the production of Vimentin mRNA expressed in mesenchymal cells. Fluorescence microscopy is used to achieve temporal detection of Vimentin mRNA in wounds. 3D light-sheet microscopy is utilized to observe the dynamic expression of Vimentin mRNA spatially around the wounded site in skin tissue. The use of DNA-gold nanoprobes to detect mRNA expression during wound healing opens up new possibilities for the study of real-time mechanisms in complex biological processes.
伤口愈合是一个高度复杂的生物学过程，伴随着细胞表型的变化、蛋白质表达的变化和活性生物分子的产生。目前，细胞内蛋白质的检测是通过免疫染色来完成的，即在固定细胞中检测标记抗体的蛋白质。然而，免疫染色不能提供关于整个组织中活细胞内动态过程的信息。在这里，提出了一种简单的方法来检测伤口愈合过程中上皮细胞向间充质细胞的转变。该方法采用 DNA 包裹的金纳米粒子荧光纳米探针来检测间充质细胞中表达的波形蛋白 mRNA。荧光显微镜用于实现对伤口中波形蛋白 mRNA 的时间检测。3D 光片显微镜用于观察皮肤组织中伤口周围 Vimentin mRNA 的动态表达。利用 DNA-金纳米探针在伤口愈合过程中检测 mRNA 的表达，为研究复杂生物过程中的实时机制开辟了新的可能性。
