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We evolved the brilliant monomeric red fluorescent protein mScarlet3 using a multiparameter screening approach. Owing to a newly engineered hydrophobic patch inside its β-barrel structure, mScarlet3 combines a high quantum yield and high fluorescence lifetime with fast and complete maturation. Consequently, mScarlet3 performs well as a fusion tag in live-cell imaging.
Integration of single-cell molecular profiling with cellular spatial localization has remained an elusive goal. Image-seq leverages high-resolution microscopy to spatially resolve and isolate viable bone marrow and leukemia cells for subsequent state-of-the art, single-cell transcriptomics.
Light-activated drugs and signaling molecules have therapeutic potential and are valuable experimental tools. Photoactivation of a mu opioid receptor agonist in the mouse brain rapidly triggered pain relief and locomotion, demonstrating that in vivo photopharmacology can drive dynamic studies into animal behavior.
A suite of tools including positive-going voltage indicators, a high-speed two-photon microscope, and denoising software enables prolonged imaging of electrical activity in neurons with limited toxicity.
Two mScarlet variants with high brightness and fast maturation times have been evolved. These variants behave favorably as fusion tags and Förster resonance energy transfer acceptors.
An international blind study confirms that smFRET measurements on dynamic proteins are highly reproducible across instruments, analysis procedures and timescales, further highlighting the promise of smFRET for dynamic structural biology.
Although structural variation is less explored than single-nucleotide variation, recent studies have shown it to be associated with several human diseases. Three fresh computational methods might help to elucidate this inadequately understood part of our genetic makeup.
This Review describes advances in cryogenic electron tomography on focused ion beam lamellae, highlighting the key benefits of this technology for in situ structural biology and discussing important future directions.
The nanopore community is stepping toward a new frontier of single-molecule protein sequencing. Here, we offer our opinions on the unique potential for this emerging technology, with a focus on single-cell proteomics, and some challenges that must be overcome to realize it.
The development of mass spectrometry-based single-cell proteomics technologies opens unique opportunities to understand the functional crosstalk between cells that drive tumor development.
