Simple super-resolution live-cell imaging based on diffusion-assisted Förster resonance energy transfer

Despite the recent development of several super-resolution fluorescence microscopic techniques, there are still few techniques that can be readily employed in conventional imaging systems. We present a very simple, rapid, general and cost-efficient super-resolution imaging method, which can be direc...

Descripción completa

Detalles Bibliográficos
Autores principales: Cho, Sangyeon, Jang, Jaeduck, Song, Chaeyeon, Lee, Heeyoung, Ganesan, Prabhakar, Yoon, Tae-Young, Kim, Mahn Won, Choi, Myung Chul, Ihee, Hyotcherl, Heo, Won Do, Park, YongKeun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3563037/
https://www.ncbi.nlm.nih.gov/pubmed/23383376
http://dx.doi.org/10.1038/srep01208
Descripción
Sumario:Despite the recent development of several super-resolution fluorescence microscopic techniques, there are still few techniques that can be readily employed in conventional imaging systems. We present a very simple, rapid, general and cost-efficient super-resolution imaging method, which can be directly employed in a simple fluorescent imaging system with general fluorophores. Based on diffusion-assisted Förster resonance energy transfer (FRET), fluorescent donor molecules that label specific target structures can be stochastically quenched by diffusing acceptor molecules, thereby temporally separating otherwise spatially overlapped fluorescence signals and allowing super-resolution imaging. The proposed method provides two- to three-fold-enhancement in spatial resolution, a significant optical sectioning property, and favorable temporal resolution in live-cell imaging. We demonstrate super-resolution live-cell dynamic imaging using general fluorophores in a standard epi-fluorescence microscope with light-emitting diode (LED) illumination. Due to the simplicity of this approach, we expect that the proposed method will prove an attractive option for super-resolution imaging.