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Live-cell protein labelling with nanometre precision by cell squeezing

January 2016. Direct observation of intracellular processes has the potential to yield insights into fundamental biological pathways and disease mechanisms. Several techniques have been developed to enable high-resolution imaging of live cells. However, the limited ability to trace intracellular components has hindered progress. Two of the persistent challenges are probe design and cellular delivery with minimal toxicity, pivotal for advances in live-cell imaging technologies. A team of scientists at Goethe University Frankfurt in collaboration with scientists at the Massachusetts Institute of Technology (MIT) in Cambridge, USA, have developed a novel approach to tag and image intracellular components in live mammalian cells. Using the microfluidic cell squeezing platform to deliver small fluorescent tris-N-nitrilotriacetic acid (trisNTA) probes (~1 kDa), the scientists demonstrated highly efficient and minimally disruptive light-triggered tracing of native proteins and subsequent super-resolution imaging of live-cell phenomena.

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.  More...





The nuclear envelope protein Lamin A can be stained selectively with the fluorecently labelled probe trisNTA (green) with the aid of the lock-and-key principle. Through the use of orthogonal labelling methods further proteins were visualised in the same cell at the same time: histone 2B (magenta), lysosomes (blue) and microtubuli (red).










Ralph Wieneke (wieneke@em.uni-frankfurt.de) and Robert Tampé (tampe@em.uni-frankfurt.de), Institute of Biochemistry, Riedberg Campus, Goethe University Frankfurt, Frankfurt am Main, Germany


Alina Kollmannsperger, Armon Sharei, Anika Raulf, Mike Heilemann, Robert Langer, Klavs F. Jensen, Ralph Wieneke & Robert Tampé: Live-cell protein labelling with nanometre precision by cell squeezing, Nature Communications 7:10372, DOI: 10.1038/ncomms10372. www.nature.com/naturecommunications. Link