OPTICS EXPRESS
·
2025
·Vol. 33
·期 9
·20023-20036
·
IF 3.3
Yuhan Nie,Yuhan Nie,Jie Hu,Siying Zhang,Xiangzhou Meng,Xiangzhou Meng,Guo Fu,Guo Fu,Guo Fu
Single-molecule localization microscopy (SMLM) breaks the diffraction limit, significantly enhancing optical resolution and providing a powerful tool for imaging complex biological structures in life sciences. However, imaging biological structures below 10 nm and studying protein-protein interactions remains a challenge for optical super-resolution microscopy. Here, we introduce a microscopy based on two-color single-molecule localization microscopy, we call it molecular Imaging by two-color imaging (MITI), which improves resolution to the molecular level, reaching several nanometers. We demonstrate MITI is suitable for samples labeled with organic dyes and photoactivatable/photoswichable fluorescent proteins. MITI can resolve two dyes separated by ssDNA with 2-10 nm length as well as ffDronpa and PAmCherry1 fused to the N- and C-terminal of FtsZ (ffDronpa-FtsZ-PAmCherry1). MITI is versatile and easy to realize, and thus, it will expand the applications of molecular scale super-resolution microscopy in life sciences.
研究领域:
生物物理学
·
细胞生物学
·
显微技术
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产品关联
DOI
PubMed
高影响因子
Nanoscale
·
2022
·Vol. 14
·期 17
·6392-6401
·
IF 8.307
Zhaoyan Yang,Yuhui Dong,Shenfei Zong,Lang Li,Kuo Yang,Zhuyuan Wang,Haibo Zeng,Yiping Cui
Single-molecule localization microscopy (SMLM) is one of the most promising super-resolution imaging techniques for visualizing ultrasmall cellular structures. Here, water-dispersed perovskite CsPbBr3 nanocrystals (CsPbBr3 NCs) fabricated by a one-step mechanochemical method are explored as a SMLM fluorophore for bioimaging. Due to their ultrahigh photoluminescence quantum yield (PLQY), inherent frequent fluorescence blinking, proper duty cycle and long-term photostability, an extremely high location precision of ∼3 nm was achieved, a sixfold enhancement than those reported previously. In addition, the spatial resolution of a SMLM image depends on the size of CsPbBr3 NCs, which is approximately 23 nm. Two closely spaced CsPbBr3 NCs with a gap of 40 nm can be clearly distinguished in the SMLM image. More importantly, unlike most perovskite quantum dots (QDs), one-step mechanochemically prepared CsPbBr3 NCs can retain their excellent fluorescence characteristics even after surface biofunctionalization, greatly reducing the current limitations of perovskite QDs on bioimaging. As an example, cell-derived exosomes (30–150 nm in diameter) labeled with CsPbBr3 NCs were easily identified by SMLM. In addition, after being functionalized with biotin, targeted SMLM imaging of the nuclear lamina or cell membranes of cells was achieved with an enhanced resolution. This work may open up a promising avenue to expand the field of perovskite QD-based SMLM to bioimaging with a high location accuracy.
研究领域:
材料科学
·
纳米技术
·
生物医学工程
·
光学
·
细胞生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ACS Applied Materials & Interfaces
·
2022
·
IF 10.383
Zhaoyan Yang,Shenfei Zong,Kuo Yang,Kai Zhu,Na Li,Zhuyuan Wang,Yiping Cui
Single molecule localization microscopy (SMLM) is indispensable in the visualization of cellular microstructures. However, current SMLM imaging materials, from organic fluorophores to quantum dots, still lack the requirement of increasing need for multiple targets of interest due to their broad emission. Here, by one-step encapsulating hydrophilic cesium lead bromide perovskite nanocrystals (CsPbBr3 NCs) into functionalized polyethylene glycol (PEG), a core–shell nanocomposite of CsPb(Cl(1–x)/Brx)3@PEG (0 < x < 1) was presented as a wavelength-tunable fluorescent probe with the narrow full width at half-maximum (fwhm) as 11 nm. The layer of functionalized PEG endows CsPbBr3 NCs with a broad spectral tunability from 521 to 431 nm, superior photostability for several years, and the ability to be further surface functionalized. The CsPb(Cl(1–x)/Brx)3@PEG exhibits a sub-10 nm localization precision and 10-fold enhanced spatial resolution. Using exosomes with small sizes less than 150 nm as the imaging target, CsPb(Cl(1–x)/Brx)3@PEG realized the distinction of two adjacent exosomes by SMLM. Moreover, after being modified with biotin, CsPb(Cl(1–x)/Brx)3@PEG was universally used for SMLM imaging of cellular microstructures. The excellent photostability and narrow fwhm indicated that such a CsPbBr3-based nanoprobe has great potential as a commercial dye for multitarget super-resolution bioimaging applications.
研究领域:
材料科学
·
纳米技术
·
生物医学工程
·
光学
·
生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）