Cargando…

Stick, stretch, and scan imaging method for DNA and filaments

Biomolecules and organelles usually undergo changes to their structure or form as a result of mechanical stretching or stimulation. It is critical to be able to observe these changes and responses, which trigger mechano-chemical coupling or signal transduction. Advanced techniques have been develope...

Descripción completa

Detalles Bibliográficos
Autores principales: Zeng, Qiuling, Gao, Yuanyuan, Yu, Hong, Zhu, Wei, Wang, Qi, Long, Quan, Fan, Zhuo, Xiao, Botao
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Royal Society of Chemistry 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9043539/
https://www.ncbi.nlm.nih.gov/pubmed/35492749
http://dx.doi.org/10.1039/d1ra07067c
_version_ 1784694903611064320
author Zeng, Qiuling
Gao, Yuanyuan
Yu, Hong
Zhu, Wei
Wang, Qi
Long, Quan
Fan, Zhuo
Xiao, Botao
author_facet Zeng, Qiuling
Gao, Yuanyuan
Yu, Hong
Zhu, Wei
Wang, Qi
Long, Quan
Fan, Zhuo
Xiao, Botao
author_sort Zeng, Qiuling
collection PubMed
description Biomolecules and organelles usually undergo changes to their structure or form as a result of mechanical stretching or stimulation. It is critical to be able to observe these changes and responses, which trigger mechano-chemical coupling or signal transduction. Advanced techniques have been developed to observe structure and form during manipulation; however, these require sophisticated methods. We have developed a simple approach to observe fine structure after stretching without fluorophore labeling. DNAs or molecules on the cell surface were bound to magnetic microbeads, followed by stretching with a magnetic field. After fixing, staining, and drying, the samples were examined by scanning electron microscopy with no need to build a functional surface with complex processes. Straight DNAs were observed rather than random-walk-like loose polymers. In our cellular experiment, the magnetic beads were bound to a Jurkat cell and formed a rosette which was later stuck to the substrate. A 41.3 μm filament on the base of a filopodium was pulled out via integrin from a cell. Therefore, our method can reveal long structures up to hundreds of micrometers at nanometer resolution after stretching or twisting. Our approach could have wide applications in structure–function studies of biomolecules, and in mechanobiology and cell biology when diffraction cannot used.
format Online
Article
Text
id pubmed-9043539
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher The Royal Society of Chemistry
record_format MEDLINE/PubMed
spelling pubmed-90435392022-04-28 Stick, stretch, and scan imaging method for DNA and filaments Zeng, Qiuling Gao, Yuanyuan Yu, Hong Zhu, Wei Wang, Qi Long, Quan Fan, Zhuo Xiao, Botao RSC Adv Chemistry Biomolecules and organelles usually undergo changes to their structure or form as a result of mechanical stretching or stimulation. It is critical to be able to observe these changes and responses, which trigger mechano-chemical coupling or signal transduction. Advanced techniques have been developed to observe structure and form during manipulation; however, these require sophisticated methods. We have developed a simple approach to observe fine structure after stretching without fluorophore labeling. DNAs or molecules on the cell surface were bound to magnetic microbeads, followed by stretching with a magnetic field. After fixing, staining, and drying, the samples were examined by scanning electron microscopy with no need to build a functional surface with complex processes. Straight DNAs were observed rather than random-walk-like loose polymers. In our cellular experiment, the magnetic beads were bound to a Jurkat cell and formed a rosette which was later stuck to the substrate. A 41.3 μm filament on the base of a filopodium was pulled out via integrin from a cell. Therefore, our method can reveal long structures up to hundreds of micrometers at nanometer resolution after stretching or twisting. Our approach could have wide applications in structure–function studies of biomolecules, and in mechanobiology and cell biology when diffraction cannot used. The Royal Society of Chemistry 2021-11-09 /pmc/articles/PMC9043539/ /pubmed/35492749 http://dx.doi.org/10.1039/d1ra07067c Text en This journal is © The Royal Society of Chemistry https://creativecommons.org/licenses/by-nc/3.0/
spellingShingle Chemistry
Zeng, Qiuling
Gao, Yuanyuan
Yu, Hong
Zhu, Wei
Wang, Qi
Long, Quan
Fan, Zhuo
Xiao, Botao
Stick, stretch, and scan imaging method for DNA and filaments
title Stick, stretch, and scan imaging method for DNA and filaments
title_full Stick, stretch, and scan imaging method for DNA and filaments
title_fullStr Stick, stretch, and scan imaging method for DNA and filaments
title_full_unstemmed Stick, stretch, and scan imaging method for DNA and filaments
title_short Stick, stretch, and scan imaging method for DNA and filaments
title_sort stick, stretch, and scan imaging method for dna and filaments
topic Chemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9043539/
https://www.ncbi.nlm.nih.gov/pubmed/35492749
http://dx.doi.org/10.1039/d1ra07067c
work_keys_str_mv AT zengqiuling stickstretchandscanimagingmethodfordnaandfilaments
AT gaoyuanyuan stickstretchandscanimagingmethodfordnaandfilaments
AT yuhong stickstretchandscanimagingmethodfordnaandfilaments
AT zhuwei stickstretchandscanimagingmethodfordnaandfilaments
AT wangqi stickstretchandscanimagingmethodfordnaandfilaments
AT longquan stickstretchandscanimagingmethodfordnaandfilaments
AT fanzhuo stickstretchandscanimagingmethodfordnaandfilaments
AT xiaobotao stickstretchandscanimagingmethodfordnaandfilaments