Fluorescence lifetime DNA-PAINT for multiplexed super-resolution imaging of cells.

Autor: Oleksiievets N; III. Institute of Physics - Biophysics, Georg August University, 37077, Göttingen, Germany., Sargsyan Y; Department of Child and Adolescent Health, University Medical Center Göttingen, 37073, Göttingen, Germany., Thiele JC; III. Institute of Physics - Biophysics, Georg August University, 37077, Göttingen, Germany., Mougios N; Institute of Neuro- and Sensory Physiology, University Medical Center Göttingen, 37073, Göttingen, Germany.; Center for Biostructural Imaging of Neurodegeneration (BIN), University Medical Center Göttingen, 37075, Göttingen, Germany., Sograte-Idrissi S; Institute of Neuro- and Sensory Physiology, University Medical Center Göttingen, 37073, Göttingen, Germany.; Center for Biostructural Imaging of Neurodegeneration (BIN), University Medical Center Göttingen, 37075, Göttingen, Germany., Nevskyi O; III. Institute of Physics - Biophysics, Georg August University, 37077, Göttingen, Germany., Gregor I; III. Institute of Physics - Biophysics, Georg August University, 37077, Göttingen, Germany., Opazo F; Institute of Neuro- and Sensory Physiology, University Medical Center Göttingen, 37073, Göttingen, Germany.; Center for Biostructural Imaging of Neurodegeneration (BIN), University Medical Center Göttingen, 37075, Göttingen, Germany.; NanoTag Biotechnologies GmbH, 37079, Göttingen, Germany., Thoms S; Department of Child and Adolescent Health, University Medical Center Göttingen, 37073, Göttingen, Germany.; Biochemistry and Molecular Medicine, Medical School, Bielefeld University, 33615, Bielefeld, Germany., Enderlein J; III. Institute of Physics - Biophysics, Georg August University, 37077, Göttingen, Germany. jenderl@gwdg.de.; Cluster of Excellence 'Multiscale Bioimaging: from Molecular Machines to Networks of Excitable Cells' (MBExC), Georg August University, Göttingen, Germany. jenderl@gwdg.de., Tsukanov R; III. Institute of Physics - Biophysics, Georg August University, 37077, Göttingen, Germany. rtsukan@gwdg.de.
Jazyk: angličtina
Zdroj: Communications biology [Commun Biol] 2022 Jan 11; Vol. 5 (1), pp. 38. Date of Electronic Publication: 2022 Jan 11.
DOI: 10.1038/s42003-021-02976-4
Abstrakt: DNA point accumulation for imaging in nanoscale topography (DNA-PAINT) is a powerful super-resolution technique highly suitable for multi-target (multiplexing) bio-imaging. However, multiplexed imaging of cells is still challenging due to the dense and sticky environment inside a cell. Here, we combine fluorescence lifetime imaging microscopy (FLIM) with DNA-PAINT and use the lifetime information as a multiplexing parameter for targets identification. In contrast to Exchange-PAINT, fluorescence lifetime PAINT (FL-PAINT) can image multiple targets simultaneously and does not require any fluid exchange, thus leaving the sample undisturbed and making the use of flow chambers/microfluidic systems unnecessary. We demonstrate the potential of FL-PAINT by simultaneous imaging of up to three targets in a cell using both wide-field FLIM and 3D time-resolved confocal laser scanning microscopy (CLSM). FL-PAINT can be readily combined with other existing techniques of multiplexed imaging and is therefore a perfect candidate for high-throughput multi-target bio-imaging.
(© 2022. The Author(s).)
Databáze: MEDLINE
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