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1.
Nano Lett ; 19(2): 1275-1281, 2019 02 13.
Artigo em Inglês | MEDLINE | ID: mdl-30681342

RESUMO

An ideal point light source is as small and as bright as possible. For fluorescent point light sources, homogeneity of the light sources is important as well as that the fluorescent units inside the light source maintain their photophysical properties, which is compromised by dye aggregation. Here we propose DNA origami as a rigid scaffold to arrange dye molecules in a dense pixel array with high control of stoichiometry and dye-dye interactions. In order to find the highest labeling density in a DNA origami structure without influencing dye photophysics, we alter the distance of two ATTO647N dyes in single base pair steps and probe the dye-dye interactions on the single-molecule level. For small distances strong quenching in terms of intensity and fluorescence lifetime is observed. With increasing distance, we observe reduced quenching and molecular dynamics. However, energy transfer processes in the weak coupling regime still have a significant impact and can lead to quenching by singlet-dark-state-annihilation. Our study fills a gap of studying the interactions of dyes relevant for superresolution microscopy with dense labeling and for single-molecule biophysics. Incorporating these findings in a 3D DNA origami object will pave the way to bright and homogeneous DNA origami nanobeads.


Assuntos
DNA/química , Corantes Fluorescentes/química , Nanoestruturas/química , Pareamento de Bases , Dimerização , Fluorescência , Microscopia Confocal , Microscopia de Fluorescência , Nanotecnologia , Espectrometria de Fluorescência
2.
Small ; 11(39): 5200-5, 2015 Oct 21.
Artigo em Inglês | MEDLINE | ID: mdl-26294348

RESUMO

A small and compact DNA cube with zeptoliter volume is constructed by means of a generalized DNA brick concept using short synthetic oligonucleotides with varying lengths. By mimicking design principles from the DNA origami technique, the DNA cube offers higher stability and assembly yields compared to other approaches. Its potential application as nanoscale fluorescent probe is demonstrated using super-resolution imaging.


Assuntos
DNA/química , DNA/ultraestrutura , Corantes Fluorescentes/síntese química , Nanopartículas/química , Nanopartículas/ultraestrutura , Tamanho da Partícula
3.
Nano Lett ; 14(3): 1627-33, 2014 Mar 12.
Artigo em Inglês | MEDLINE | ID: mdl-24517269

RESUMO

The combination of molecular self-assembly based on the DNA origami technique with lithographic patterning enables the creation of hierarchically ordered nanosystems, in which single molecules are positioned at precise locations on multiple length scales. Based on a hybrid assembly protocol utilizing DNA self-assembly and electron-beam lithography on transparent glass substrates, we here demonstrate a DNA origami microarray, which is compatible with the requirements of single molecule fluorescence and super-resolution microscopy. The spatial arrangement allows for a simple and reliable identification of single molecule events and facilitates automated read-out and data analysis. As a specific application, we utilize the microarray to characterize the performance of DNA strand displacement reactions localized on the DNA origami structures. We find considerable variability within the array, which results both from structural variations and stochastic reaction dynamics prevalent at the single molecule level.


Assuntos
DNA/química , Conformação de Ácido Nucleico , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Microscopia de Fluorescência , Análise de Sequência com Séries de Oligonucleotídeos/instrumentação
4.
Angew Chem Int Ed Engl ; 53(29): 7665-8, 2014 Jul 14.
Artigo em Inglês | MEDLINE | ID: mdl-24894973

RESUMO

The arrangement of DNA-based nanostructures into extended higher order assemblies is an important step towards their utilization as functional molecular materials. We herein demonstrate that by electrostatically controlling the adhesion and mobility of DNA origami structures on mica surfaces by the simple addition of monovalent cations, large ordered 2D arrays of origami tiles can be generated. The lattices can be formed either by close-packing of symmetric, non-interacting DNA origami structures, or by utilizing blunt-end stacking interactions between the origami units. The resulting crystalline lattices can be readily utilized as templates for the ordered arrangement of proteins.


Assuntos
DNA/química , Microscopia de Força Atômica , Conformação de Ácido Nucleico , Eletricidade Estática
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