Abstract
The molecular machinery of life is founded on chiral building blocks, but no experimental technique is currently available to distinguish or monitor chiral systems in live cell bio-imaging studies. Luminescent chiral molecules encode a unique optical fingerprint within emitted circularly polarized light (CPL) carrying information about the molecular environment, conformation, and binding state. Here, we present a CPL Laser Scanning Confocal Microscope (CPL-LSCM) capable of simultaneous chiroptical contrast based live-cell imaging of endogenous and engineered CPL-active cellular probes. Further, we demonstrate that CPL-active probes can be activated using two-photon excitation, with complete CPL spectrum recovery. The combination of these two milestone results empowers the multidisciplinary imaging community, allowing the study of chiral interactions on a sub-cellular level in a new (chiral) light.
Highlights
The molecular machinery of life is founded on chiral building blocks, but no experimental technique is currently available to distinguish or monitor chiral systems in live cell bioimaging studies
circularly polarized light (CPL) spectroscopy is a well-established discipline, with CPL emission maximised for emission transitions which are magnetic dipole (MD) allowed and electric dipole (ED) forbidden
We have demonstrated the CPL-LSCM system, which is capable of rapidly and simultaneously acquiring diffraction-limited enantioselective chiral-contrast based CPLdifferential images for the sub-cellular tracking of emissive chiral sub-cellular probes. This milestone is accompanied by the proofof-concept demonstration of 2PE-CPL spectroscopy, showing that low-energy 2PE-CPL-LSCM is ripe for future development
Summary
The molecular machinery of life is founded on chiral building blocks, but no experimental technique is currently available to distinguish or monitor chiral systems in live cell bioimaging studies. To advance CPL microscopy, we have previously built a proof-ofconcept chiroptical contrast time-resolved epifluorescence microscope and demonstrated enantioselective differential chiral contrast (EDCC) imaging, examining the Λ- and Δ-enantiomers of a bright
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