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

2022-01-11 | journal article; research paper. A publication with affiliation to the University of Göttingen.

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​Fluorescence lifetime DNA-PAINT for multiplexed super-resolution imaging of cells​
Oleksiievets, N.; Sargsyan, Y.; Thiele, J. C. ; Mougios, N.; Sograte-Idrissi, S.; Nevskyi, O. & Gregor, I. et al.​ (2022) 
Communications Biology5(1) pp. 38​.​ DOI: https://doi.org/10.1038/s42003-021-02976-4 

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Authors
Oleksiievets, Nazar; Sargsyan, Yelena; Thiele, Jan Christoph ; Mougios, Nikolaos; Sograte-Idrissi, Shama; Nevskyi, Oleksii; Gregor, Ingo; Opazo, Felipe; Thoms, Sven ; Enderlein, Jörg ; Tsukanov, Roman
Abstract
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.
Issue Date
11-January-2022
Journal
Communications Biology 
Project
EXC 2067: Multiscale Bioimaging 
SFB 1002: Modulatorische Einheiten bei Herzinsuffizienz 
SFB 1002 | A10: Peroxisomen als modulatorische Einheiten im Herzstoffwechsel und bei Herzinsuffizienz 
Organization
III. Physikalisches Institut - Biophysik 
Working Group
RG Enderlein 
RG Thoms (Biochemistry and Molecular Medicine) 
ISSN
2399-3642
Language
English
Sponsor
Open-Access-Publikationsfonds 2022

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