Longfang Yao , Li Zhang , Liwen Chen , Yiyan Fei , Simone Lamon , Min Gu , Lan Mi , Jing Wang , Jiong Ma
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引用次数: 0
摘要
膨胀显微镜(ExM)是一种广泛使用的超分辨率技术,可对光衍射极限以外的结构成像。然而,ExM 存在标记信号弱和膨胀失真等问题,限制了其适用性。在这里,我们提出了一种名为四面体DNA纳米结构膨胀显微镜(TDN-ExM)的创新方法,通过使用四面体DNA纳米结构(TDNs)进行荧光标记来解决这些局限性。由于四维 DNA 纳米结构具有多维性,允许对多种染料进行修饰,因此我们的方法可将信号放大 3 到 10 倍。以前的研究已经证实,大范围的失真极小,而我们的策略可以减少样品超微结构层面的失真,因为它不依赖锚定剂,也不受消化的影响。这使得荧光更明亮、均匀度更高,并与不同的标记策略和光学超分辨率技术兼容。我们通过对各种生物结构进行成像,提高了分辨率和信噪比,从而验证了 TDN-ExM 的实用性。
Visualizing highly bright and uniform cellular ultrastructure by expansion-microscopy with tetrahedral DNA nanostructures
Expansion Microscopy (ExM) is a widely used super-resolution technique that enables imaging of structures beyond the diffraction limit of light. However, ExM suffers from weak labeling signals and expansion distortions, limiting its applicability. Here, we present an innovative approach called Tetrahedral DNA nanostructure Expansion Microscopy (TDN-ExM), addressing these limitations by using tetrahedral DNA nanostructures (TDNs) for fluorescence labeling. Our approach demonstrates a 3- to 10-fold signal amplification due to the multivertex nature of TDNs, allowing the modification of multiple dyes. Previous studies have confirmed minimal distortion on a large scale, and our strategy can reduce the distortion at the ultrastructural level in samples because it does not rely on anchoring agents and is not affected by digestion. This results in a brighter fluorescence, better uniformity, and compatibility with different labeling strategies and optical super-resolution technologies. We validated the utility of TDN-ExM by imaging various biological structures with improved resolutions and signal-to-noise ratios.
期刊介绍:
The Journal of Photochemistry and Photobiology B: Biology provides a forum for the publication of papers relating to the various aspects of photobiology, as well as a means for communication in this multidisciplinary field.
The scope includes:
- Bioluminescence
- Chronobiology
- DNA repair
- Environmental photobiology
- Nanotechnology in photobiology
- Photocarcinogenesis
- Photochemistry of biomolecules
- Photodynamic therapy
- Photomedicine
- Photomorphogenesis
- Photomovement
- Photoreception
- Photosensitization
- Photosynthesis
- Phototechnology
- Spectroscopy of biological systems
- UV and visible radiation effects and vision.