Live-cell protein labelling with nanometre precision by cell squeezing.

Area of Science:

• Cell biology
• Microscopy
• Biotechnology

Background:

• Current live-cell labelling methods have limitations in efficiency, specificity, and cell permeability.
• Visualizing proteins in live cells with minimal disturbance is crucial for understanding cellular processes.

Purpose of the Study:

• To develop a high-throughput, efficient, and specific method for live-cell protein labelling.
• To enable super-resolution imaging of proteins in various subcellular compartments.

Main Methods:

• Utilized minimalistic probes delivered via microfluidic cell squeezing for high-throughput protein labelling in mammalian cells.
• Demonstrated high-affinity and target-specific tracing of proteins.
• Achieved photoinduced labelling within live cells.

Main Results:

• Enabled fine-tuned delivery of subnanomolar probe concentrations with minimal probe size.
• Facilitated live-cell super-resolution imaging with low background and nanometre precision.
• Achieved rapid probe delivery (∼1,000,000 cells per second) across diverse cell types.

Conclusions:

• The developed technique offers a versatile and efficient approach for live-cell protein visualization.
• This method shows potential for precise tracing of target proteins in live mammalian cells using super-resolution microscopy.
• The technique is compatible with established methods for multiplexed labelling.