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Sensing of vimentin mRNA in 2D and 3D models of wounded skin using DNA-coated gold nanoparticles

Sensing of vimentin mRNA in 2D and 3D models of wounded skin using DNA-coated gold nanoparticles
Sensing of vimentin mRNA in 2D and 3D models of wounded skin using DNA-coated gold nanoparticles
Wound healing is a highly complex biological process, which is accompanied by changes in cell phenotype, variations in protein expression, and the production of active biomolecules. Currently, the detection of proteins in cells is done by immunostaining where the proteins in fixed cells are detected by labeled antibodies. However, immunostaining cannot provide information about dynamic processes in living cells, within the whole tissue. Here, an easy method is presented to detect the transition of epithelial to mesenchymal cells during wound healing. The method employs DNA-coated gold nanoparticle fluorescent nanoprobes to sense the production of Vimentin mRNA expressed in mesenchymal cells. Fluorescence microscopy is used to achieve temporal detection of Vimentin mRNA in wounds. 3D light-sheet microscopy is utilized to observe the dynamic expression of Vimentin mRNA spatially around the wounded site in skin tissue. The use of DNA–gold nanoprobes to detect mRNA expression during wound healing opens up new possibilities for the study of real-time mechanisms in complex biological processes.
1613-6810
Vilela, Patrick
fd15dd30-ae72-40b8-8aac-546fa2bf54f5
Heuer-jungemann, Amelie
48f01a95-9ba4-4248-bd24-0e20d601d2cc
El-Sagheer, Afaf
05b8295a-64ad-4fdf-ad57-c34934a46c04
Brown, Tom
a64aae36-bb30-42df-88a2-11be394e8c89
Muskens, Otto T.
2284101a-f9ef-4d79-8951-a6cda5bfc7f9
Smyth, Neil R.
0eba2a40-3b43-4d40-bb64-621bd7e9d505
Kanaras, Antonios G.
667ecfdc-7647-4bd8-be03-a47bf32504c7
Vilela, Patrick
fd15dd30-ae72-40b8-8aac-546fa2bf54f5
Heuer-jungemann, Amelie
48f01a95-9ba4-4248-bd24-0e20d601d2cc
El-Sagheer, Afaf
05b8295a-64ad-4fdf-ad57-c34934a46c04
Brown, Tom
a64aae36-bb30-42df-88a2-11be394e8c89
Muskens, Otto T.
2284101a-f9ef-4d79-8951-a6cda5bfc7f9
Smyth, Neil R.
0eba2a40-3b43-4d40-bb64-621bd7e9d505
Kanaras, Antonios G.
667ecfdc-7647-4bd8-be03-a47bf32504c7

Vilela, Patrick, Heuer-jungemann, Amelie, El-Sagheer, Afaf, Brown, Tom, Muskens, Otto T., Smyth, Neil R. and Kanaras, Antonios G. (2018) Sensing of vimentin mRNA in 2D and 3D models of wounded skin using DNA-coated gold nanoparticles. Small. (doi:10.1002/smll.201703489).

Record type: Article

Abstract

Wound healing is a highly complex biological process, which is accompanied by changes in cell phenotype, variations in protein expression, and the production of active biomolecules. Currently, the detection of proteins in cells is done by immunostaining where the proteins in fixed cells are detected by labeled antibodies. However, immunostaining cannot provide information about dynamic processes in living cells, within the whole tissue. Here, an easy method is presented to detect the transition of epithelial to mesenchymal cells during wound healing. The method employs DNA-coated gold nanoparticle fluorescent nanoprobes to sense the production of Vimentin mRNA expressed in mesenchymal cells. Fluorescence microscopy is used to achieve temporal detection of Vimentin mRNA in wounds. 3D light-sheet microscopy is utilized to observe the dynamic expression of Vimentin mRNA spatially around the wounded site in skin tissue. The use of DNA–gold nanoprobes to detect mRNA expression during wound healing opens up new possibilities for the study of real-time mechanisms in complex biological processes.

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revised_Kanaras_manuscript_skin (1) - Accepted Manuscript
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Accepted/In Press date: 11 January 2018
e-pub ahead of print date: 21 February 2018

Identifiers

Local EPrints ID: 418278
URI: https://eprints.soton.ac.uk/id/eprint/418278
ISSN: 1613-6810
PURE UUID: dab4f785-184a-4d5c-8ba3-5ae444c73ed2
ORCID for Otto T. Muskens: ORCID iD orcid.org/0000-0003-0693-5504
ORCID for Antonios G. Kanaras: ORCID iD orcid.org/0000-0002-9847-6706

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Date deposited: 27 Feb 2018 17:30
Last modified: 15 Aug 2019 04:20

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Contributors

Author: Patrick Vilela
Author: Amelie Heuer-jungemann
Author: Afaf El-Sagheer
Author: Tom Brown
Author: Otto T. Muskens ORCID iD
Author: Neil R. Smyth

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