Fluorescence lifetime DNA-PAINT for multiplexed super-resolution imaging of cells.
Commun Biol
; 5(1): 38, 2022 01 11.
Article
em En
| MEDLINE
| ID: mdl-35017652
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.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
DNA
/
Nanotecnologia
/
Imagem Molecular
/
Microscopia de Fluorescência
Idioma:
En
Ano de publicação:
2022
Tipo de documento:
Article