Reductively Caged, Photoactivatable DNA‐PAINT for High‐Throughput Super‐resolution Microscopy

2020 ◽  
Vol 59 (29) ◽  
pp. 11758-11762
Author(s):  
Soohyun Jang ◽  
Mingi Kim ◽  
Sang‐Hee Shim
Keyword(s):  
High Throughput ◽  
Super Resolution ◽  
Super Resolution Microscopy

scholarly journals Photonic chip-based multimodal super-resolution microscopy for histopathological assessment of cryopreserved tissue sections

2021 ◽  
Author(s):  
Luis E. Villegas-Hernandez ◽  
Vishesh Kumar Dubey ◽  
Mona Nystad ◽  
Jean-Claude Tinguely ◽  
David A. Coucheron ◽  
...  
Keyword(s):  
High Throughput ◽  
Single Molecule ◽  
Operating Cost ◽  
Super Resolution ◽  
Anatomical Variations ◽  
Histopathological Analysis ◽  
Tissue Samples ◽  
Tissue Sections ◽  
Imaging Tool ◽  
Super Resolution Microscopy

Histopathological assessment involves the identification of anatomical variations in tissues that are associated with diseases. While diffraction-limited optical microscopes assist in the diagnosis of a wide variety of pathologies, their resolving capabilities are insufficient to visualize some anomalies at subcellular level. Although a novel set of super-resolution optical microscopy techniques can fulfill the resolution demands in such cases, the system complexity, high operating cost, lack of multimodality, and low-throughput imaging of these methods limit their wide adoption in clinical settings. In this study, we interrogate the photonic chip as an attractive high-throughput super-resolution microscopy platform for histopathology. Using cryopreserved ultrathin tissue sections of human placenta, mouse kidney, and zebrafish eye retina prepared by the Tokuyasu method, we validate the photonic chip as a multi-modal imaging tool for histo-anatomical analysis. We demonstrate that photonic-chip platform can deliver multi-modal imaging capabilities such as total internal reflection fluorescence microscopy, intensity fluctuation-based optical nanoscopy, single-molecule localization microscopy, and correlative light-electron microscopy. Our results demonstrate that the photonic chip-based super-resolution microscopy platform has the potential to deliver high-throughput multimodal histopathological analysis of cryopreserved tissue samples.

scholarly journals High throughput 3D super-resolution microscopy reveals Caulobacter crescentus in vivo Z-ring organization

2014 ◽  
Vol 111 (12) ◽  
pp. 4566-4571 ◽  
Author(s):  
S. J. Holden ◽  
T. Pengo ◽  
K. L. Meibom ◽  
C. Fernandez Fernandez ◽  
J. Collier ◽  
...  
Keyword(s):  
High Throughput ◽  
Caulobacter Crescentus ◽  
Super Resolution ◽  
Z Ring ◽  
Super Resolution Microscopy

scholarly journals AN ECONOMIC, SQUARE-SHAPED FLAT-FIELD ILLUMINATION MODULE FOR TIRF-BASED SUPER-RESOLUTION MICROSCOPY

2021 ◽  
Author(s):  
Jeff Y L Lam ◽  
Yunzhao Wu ◽  
Eleni Dimou ◽  
Ziwei Zhang ◽  
Matthew R Cheetham ◽  
...  
Keyword(s):  
High Throughput ◽  
Total Internal Reflection ◽  
Quantitative Imaging ◽  
Super Resolution ◽  
Cost Effective ◽  
Cellular Systems ◽  
Cost Effective Method ◽  
Flat Field ◽  
Super Resolution Microscopy ◽  
Highly Uniform

Super-resolution (SR) microscopy allows complex biological assemblies to be observed with remarkable resolution. However, the presence of uneven Gaussian-shaped illumination hinders its use in quantitative imaging or high-throughput assays. Methods developed to circumvent this problem are often expensive, hard-to-implement, or not applicable to total internal reflection fluorescence (TIRF) imaging. We herein demonstrate a cost-effective method to overcome these challenges using a small square-core multimodal optical fibre as the coupler. We characterise our method with synthetic, recombinant and cellular systems imaged under TIRF and highly inclined and laminated optical sheet (HILO) illuminations to demonstrate its ability to produce highly uniform images under all conditions.

scholarly journals A novel GSK3-regulated APC:Axin interaction regulates Wnt signaling by driving a catalytic cycle of efficient βcatenin destruction

eLife ◽  
2015 ◽  
Vol 4 ◽  
Author(s):  
Mira I Pronobis ◽  
Nasser M Rusan ◽  
Mark Peifer
Keyword(s):  
Wnt Signaling ◽  
Dynamic Model ◽  
High Throughput ◽  
Mammalian Cells ◽  
Super Resolution ◽  
Negative Regulator ◽  
Functional Tests ◽  
Interaction Site ◽  
Signaling Complex ◽  
Super Resolution Microscopy

APC, a key negative regulator of Wnt signaling in development and oncogenesis, acts in the destruction complex with the scaffold Axin and the kinases GSK3 and CK1 to target βcatenin for destruction. Despite 20 years of research, APC's mechanistic function remains mysterious. We used FRAP, super-resolution microscopy, functional tests in mammalian cells and flies, and other approaches to define APC's mechanistic role in the active destruction complex when Wnt signaling is off. Our data suggest APC plays two roles: (1) APC promotes efficient Axin multimerization through one known and one novel APC:Axin interaction site, and (2) GSK3 acts through APC motifs R2 and B to regulate APC:Axin interactions, promoting high-throughput of βcatenin to destruction. We propose a new dynamic model of how the destruction complex regulates Wnt signaling and how this goes wrong in cancer, providing insights into how this multiprotein signaling complex is assembled and functions via multivalent interactions.

Reductively Caged, Photoactivatable DNA‐PAINT for High‐Throughput Super‐resolution Microscopy

2020 ◽  
Vol 132 (29) ◽  
pp. 11856-11860
Author(s):  
Soohyun Jang ◽  
Mingi Kim ◽  
Sang‐Hee Shim
Keyword(s):  
High Throughput ◽  
Super Resolution ◽  
Super Resolution Microscopy

scholarly journals Metrology of DNA arrays by super-resolution microscopy

Nanoscale ◽  
2017 ◽  
Vol 9 (29) ◽  
pp. 10205-10211 ◽  
Author(s):  
Christopher M. Green ◽  
Kelly Schutt ◽  
Noah Morris ◽  
Reza M. Zadegan ◽  
William L. Hughes ◽  
...  
Keyword(s):  
Quantitative Analysis ◽  
High Throughput ◽  
Self Assembly ◽  
Super Resolution ◽  
Dna Nanostructures ◽  
Dna Arrays ◽  
Resolution Imaging ◽  
Super Resolution Microscopy

Crystal-PAINT super-resolution imaging enables high-throughput metrology of DNA nanostructures for quantitative analysis of arrays formed through self-assembly.

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