Single-molecule localization microscopy(SMLM)surpasses the diffraction limit by randomly switching fluorophores between fluorescent and dark states,precisely pinpointing the resulted isolated emission patterns,thereby...Single-molecule localization microscopy(SMLM)surpasses the diffraction limit by randomly switching fluorophores between fluorescent and dark states,precisely pinpointing the resulted isolated emission patterns,thereby reconstructing the superresolution images based on the accumulated locations of thousands to millions of single molecules.This technique achieves a ten-fold improvement in resolution,unveiling the intricate details of molecular activities and structures in cells and tissues.Multicolor SMLM extends this capability by imaging distinct protein species labeled with various fluorescent probes,providing insights into structural intricacies and spatial relationships among different targets.This review explores recent advancements in multicolor SMLM,evaluates the strengths and limitations of each variant,and discusses the future prospects.展开更多
基金funded by the Natural Science Foundation of Liaoning Province(2023-MS-103),and the National Natural Science Foundation of China(62305041).
文摘Single-molecule localization microscopy(SMLM)surpasses the diffraction limit by randomly switching fluorophores between fluorescent and dark states,precisely pinpointing the resulted isolated emission patterns,thereby reconstructing the superresolution images based on the accumulated locations of thousands to millions of single molecules.This technique achieves a ten-fold improvement in resolution,unveiling the intricate details of molecular activities and structures in cells and tissues.Multicolor SMLM extends this capability by imaging distinct protein species labeled with various fluorescent probes,providing insights into structural intricacies and spatial relationships among different targets.This review explores recent advancements in multicolor SMLM,evaluates the strengths and limitations of each variant,and discusses the future prospects.
