Fluorescent Silica Nanoparticles with Well-Separated Intensity Distributions from Batch Reactions.

Silica chemistry provides pathways to uniquely tunable nanoparticle platforms for biological imaging. It has been a long-standing problem to synthesize fluorescent silica nanoparticles (SNPs) in batch reactions with high and low fluorescence intensity levels for reliable use as an intensity barcode, which would greatly increase the number of molecular species that could be tagged intracellularly and simultaneously observed in conventional fluorescence microscopy. Here, employing an amino-acid catalyzed growth, highly fluorescent SNP probes were synthesized with sizes <40 nm and well-separated intensity distributions, as mapped by single-particle imaging techniques. A seeded growth approach was used to minimize the rate of secondary particle formation. Organic fluorescent dye affinity for the SNP during shell growth was tuned using specifics of the organosilane linker chemistry. This work highlights design considerations in the development of fluorescent probes with well-separated intensity distributions synthesized in batch reactions for single-particle imaging and sensing applications, where heterogeneities across the nanoparticle ensemble are critical factors in probe performance.

A Labeling Strategy for Living Specimens in Long-Term/Super-Resolution Fluorescence Imaging.

Despite the urgent need to image living specimens for cutting-edge biological research, most existing fluorescent labeling methods suffer from either poor optical properties or complicated operations required to realize cell-permeability and specificity. In this study, we introduce a method to overcome these limits-taking advantage of the intrinsic affinity of bright and photostable fluorophores, no matter if they are supposed to be live-cell incompatible or not. Incubated with living cells and tissues in particular conditions (concentration and temperature), some Atto and BODIPY dyes show live-cell labeling capability for specific organelles without physical cell-penetration or chemical modifications. Notably, by using Atto 647N as a live-cell mitochondrial marker, we obtain 2.5-time enhancement of brightness and photostability compared with the most commonly used SiR dye in long-term imaging. Our strategy has expanded the scientist's toolbox for understanding the dynamics and interactions of subcellular structures in living specimens.

Dye-Doped Fluorescent Silica Nanoparticles for Live Cell and In Vivo Bioimaging.

The need for novel design strategies for fluorescent nanomaterials to improve our understanding of biological activities at the molecular level is increasing rapidly. Dye-doped fluorescent silica nanoparticles (SiNPs) emerge with great potential for developing fluorescence imaging techniques as a novel and ideal platform for the monitoring of living cells and the whole body. Organic dye-containing fluorescent SiNPs exhibit many advantages: they have excellent biocompatibility, are non-toxic, highly hydrophilic, optically transparent, size-tunable and easily modified with various biomolecules. The outer silica shell matrix protects fluorophores from outside chemical reaction factors and provides a hydrophilic shell for the insoluble nanoparticles, which enhances the photo-stability and biocompatibility of the organic fluorescent dyes. Here, we give a summary of the synthesis, characteristics and applications of fluorescent SiNPs for non-invasive fluorescence bioimaging in live cells and in vivo. Additionally, the challenges and perspectives of SiNPs are also discussed. We prospect that the further development of these nanoparticles will lead to an exciting breakthrough in the understanding of biological processes.