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2022 ◽  
Vol 19 (2) ◽  
pp. 026001
Author(s):  
N A Smirnov ◽  
S I Kudryashov ◽  
А А Rudenko ◽  
A A Nastulyavichus ◽  
A A Ionin

Abstract A comparison of single-pulse laser ablation of gold target by pulses with a 0.3–10 ps duration and a wavelength of 515 nm in air and in water was performed. The radiation was focused on the sample surface through the objectives with numerical apertures NA = 0.65 and 0.25. The influence of the medium, pulse duration, and spot size on the crater morphology was studied. A significant difference in crater morphology was found for different lenses. The ablation efficiency was studied by measuring the profiles of single-shot pulse craters using scanning force microscopy. The contribution of filamentation to the ablation process is shown quantitatively.


eLife ◽  
2021 ◽  
Vol 10 ◽  
Author(s):  
Maarten W Paul ◽  
Arshdeep Sidhu ◽  
Yongxin Liang ◽  
Sarah E van Rossum-Fikkert ◽  
Hanny Odijk ◽  
...  

BRCA2 is an essential protein in genome maintenance, homologous recombination and replication fork protection. Its function includes multiple interaction partners and requires timely localization to relevant sites in the nucleus. We investigated the importance of the highly conserved DNA binding domain (DBD) and C-terminal domain (CTD) of BRCA2. We generated BRCA2 variants missing one or both domains in mouse ES cells and defined their contribution in HR function and dynamic localization in the nucleus, by single particle tracking of BRCA2 mobility. Changes in molecular architecture of BRCA2 induced by binding partners of purified BRCA2 was determined by scanning force microscopy. BRCA2 mobility and DNA damage-induced increase in the immobile fraction was largely unaffected by C-terminal deletions. The purified proteins missing CTD and/or DBD were defective in architectural changes correlating with reduced homologous recombination function in cells. These results emphasize BRCA2 activity at sites of damage beyond promoting RAD51 delivery.


2021 ◽  
Author(s):  
Maarten W. Paul ◽  
Arshdeep Sidhu ◽  
Yongxin Liang ◽  
Sarah E. van Rossum-Fikkert ◽  
Hanny Odijk ◽  
...  

AbstractBRCA2 is an essential protein in genome maintenance, homologous recombination and replication fork protection. Its function includes multiple interaction partners and requires timely localization to relevant sites in the nucleus. We investigated the importance of the highly conserved DNA binding domain (DBD) and C-terminal domain (CTD) of BRCA2. We generated BRCA2 variants missing one or both domains in mouse ES cells and defined their contribution in HR function and dynamic localization in the nucleus, by single particle tracking of BRCA2 mobility. Changes in molecular architecture of BRCA2 induced by binding partners of purified BRCA2 was determined by scanning force microscopy. BRCA2 mobility and DNA damage-induced increase in the immobile fraction was largely unaffected by C- terminal deletions. The purified proteins missing CTD and/or DBD were defective in architectural changes correlating with reduced homologous recombination function in cells. These results emphasize BRCA2 activity at sites of damage beyond promoting RAD51 delivery.


2021 ◽  
Vol 15 (2) ◽  
Author(s):  
David Hälg ◽  
Thomas Gisler ◽  
Yeghishe Tsaturyan ◽  
Letizia Catalini ◽  
Urs Grob ◽  
...  

2020 ◽  
Vol 32 (3) ◽  
pp. 035706
Author(s):  
Philipp Wagner ◽  
Adam Foster ◽  
Insook Yi ◽  
Masayuki Abe ◽  
Yoshiaki Sugimoto ◽  
...  

2020 ◽  
Author(s):  
Congzhou Chen ◽  
Jin Xu ◽  
Xiaolong Shi

Abstract Background: DNA origami can be applied as a “ruler” for nanoscale calibration or super-resolution fluorescence microscopy with an ideal structure for defining fluorophore arrangement, allowing the distance between fluorophores to be precisely controlled at the nanometer scale. DNA origami can also be used as a nanotag with arbitrary programmable shapes. Result: We formed a hexagonal origami structure embedded with three different fluorescent dyes on the surface. The distance between each fluorescent block was ~120 nm, which is below the diffraction limit of light, allowing for its application as a nano-ruler for super-resolution fluorescence microscopy. The outside edge of the hexagonal structure was redesigned to form three different substructures as topological labels. Atomic and scanning force microscopy demonstrated consistency of the nanoscale distance between morphological and fluorescent labels. Conclusion: We assembled the hexagonal origami platform and confirmed the fluorescent and topological lables, this fluorophore-embedded hexagonal origami platform can be used as a dual nano-ruler for both optical and topological calibration.


Polymers ◽  
2020 ◽  
Vol 12 (9) ◽  
pp. 2118
Author(s):  
Takanori Goto ◽  
Jun Morita ◽  
Yuya Maekawa ◽  
Shinji Kanehashi ◽  
Takeshi Shimomura

We investigated the electrical properties of a composite film loaded with semi-conductive poly(3-hexylthiophene) (P3HT) nanofibers dispersed in poly(styrene-b-butadiene-b-styrene) (SBS). This structure can be regarded as the hybrid of SBS matrix with elastic mechanical properties and P3HT nanofibers with semiconducting properties. The P3HT nanofibers were embedded in the fingerprint pattern of microphase-separated SBS, as observed by scanning force microscopy. Furthermore, the electrical conductivity and field-effect mobility of the composite films were evaluated. The field-effect mobility was estimated to be 6.96 × 10−3 cm2 V−1 s−1, which is consistent with the results of previous studies on P3HT nanofibers dispersed in an amorphous polymer matrix including poly(methyl methacrylate) and polystyrene, and we found that the P3HT nanofiber network was connected in the SBS bulk matrix. The film was stretchable; however, at elongation by two times, the nanofiber network could not follow the elongation of the SBS matrix, and the conductivity decreased drastically. The field-effect transistor of this film was operated by bending deformation with a radius of curvature of 1.75 cm, though we could not obtain an off-state and the device operated in a normally-on state.


2020 ◽  
Vol 48 (17) ◽  
pp. 9649-9659 ◽  
Author(s):  
Arshdeep Sidhu ◽  
Małgorzata Grosbart ◽  
Humberto Sánchez ◽  
Bram Verhagen ◽  
Nick L L van der Zon ◽  
...  

Abstract BRCA2 is a key breast cancer associated protein that is predicted to have interspersed regions of intrinsic disorder. Intrinsic disorder coupled with large size likely allows BRCA2 to sample a broad range of conformational space. We expect that the resulting dynamic arrangements of BRCA2 domains are a functionally important aspect of its role in homologous recombination DNA repair. To determine the architectural organization and the associated conformational landscape of BRCA2, we used scanning force microscopy based single molecule analyses to map the flexible regions of the protein and characterize which regions influence oligomerization. We show that the N- and the C-terminal regions are the main flexible regions. Both of these regions also influence BRCA2 oligomerization and interaction with RAD51. In the central Brc repeat region, Brc 1–4 and Brc 5–8 contribute synergistically to BRCA2 interaction with RAD51. We also analysed several single amino acid changes that are potentially clinically relevant and found one, the variant of F1524V, which disrupts key interactions and alters the conformational landscape of the protein. We describe the overall conformation spectrum of BRCA2, which suggests that dynamic structural transitions are key features of its biological function, maintaining genomic stability.


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