Extra kinetic dimensions for label discrimination

• DOI: 10.1038/s41467-022-29172-0
• Publication Date: 2022-03-18T11:12:28Z
• Authors: Raja Chouket; Agnès Pellissier-...; Aliénor Lahlou; Ruikang Zhang; Diana Kim; Marie-Aude Plamont; Mingshu Zhang; Xi Zhang; Pingyong Xu; Nicolas Desprat; Dominique Bourgeois; Agathe Espagne; Annie Lemarchand; Thomas Le Saux; Ludovic Jullien
• Keywords: Biomolecule; Chromatic aberration

Due to its sensitivity and versatility, fluorescence is widely used detect specifically labeled biomolecules. However, currently limited by label discrimination, which suffers from the broad full width of absorption/emission bands narrow lifetime distribution bright fluorophores. We overcome this limitation introducing extra kinetic dimensions through illuminations reversibly photoswitchable fluorophores (RSFs) at different light intensities. In expanded space, each RSF characterized a chromatic aberration-free fingerprint photochemical reactivity, can be recovered with hardware, excellent photon budget, minimal data processing. This was identify discriminate up 20 among 22 spectrally similar fluorescent proteins (RSFPs) in less than 1s. strategy opens promising perspectives for expanding multiplexing capabilities imaging.