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

Abstract

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.

Phanta: A Non-Fluorescent Photochromic Acceptor for pcFRET

“…Phanta will allow increased multiplexing of high-throughput FRET-based assays designed around fluorescence plate readers. Phanta has the potential to improve new super-resolution approaches that rely on FRET [29] by allowing dual channel super-resolution imaging or providing a way to tune fluorescence fluctuation by altering FRET efficiency through photoswitching.…”

Single-molecule fluorescence resonance energy transfer in molecular biology

“…137 Super-resolution based smFRET is also reported recently to get high spatiotemporal resolution. 138 …”

AIE Based Coumarin Chromophore - Evaluation and Correlation Between Solvatochromism and Solvent Polarity Parameters

“…Coumarin constitute the largest class of laser dyes in the 'blue-green' region [21][22][23][24][25]. Coumarin dyes have also been used as blue, green and red dopants in organic light-emitting diodes (OLEDs) [26][27][28], in dye-sensitized solar cells (DSSC) [29][30][31] and tailored to in vivo imaging applications [32][33][34][35]. In recent years Wang and co-workers report AIE active coumarin-based Schiff bases and successively put into applications in cell imaging [36].…”