Live-cell labelling techniques to visualize proteins with minimal disturbance are important; however, the currently available methods are limited in their labelling efficiency, specificity and cell permeability. We describe high-throughput protein labelling facilitated by minimalistic probes delivered to mammalian cells by microfluidic cell squeezing. High-affinity and target-specific tracing of proteins in various subcellular compartments is demonstrated, culminating in photoinduced labelling within live cells. Both the fine-tuned delivery of subnanomolar concentrations and the minimal size of the probe allow for live-cell super-resolution imaging with very low background and nanometre precision. This method is fast in probe delivery (∼ 1,000,000 cells per second), versatile across cell types and can be readily transferred to a multitude of proteins. Moreover, the technique succeeds in combination with well-established methods to gain multiplexed labelling and has demonstrated potential to precisely trace target proteins, in live mammalian cells, by super-resolution microscopy.

中文翻译：

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通过细胞挤压以纳米精度对活细胞蛋白质进行标记。

活细胞标记技术以最小的干扰来可视化蛋白质非常重要；然而，目前可用的方法在其标记效率，特异性和细胞渗透性方面受到限制。我们描述了通过微流体细胞挤压传递到哺乳动物细胞的简约探针促进的高通量蛋白质标记。证明了在各种亚细胞区室中蛋白质的高亲和力和靶标特异性追踪，最终导致了活细胞内的光诱导标记。亚纳摩尔浓度的微调递送和探针的最小尺寸都允许以非常低的背景和纳米精度进行活细胞超分辨率成像。这种方法的探针传输速度非常快（每秒约1,000,000个细胞），跨细胞类型通用，可以很容易地转移到多种蛋白质上。此外，该技术成功地与成熟的方法相结合，获得了多重标记，并已显示出通过超分辨率显微镜在活的哺乳动物细胞中精确追踪目标蛋白的潜力。