3D immuno-confocal image reconstruction of fibroblast cytoskeleton and nucleus architecture.
3D reconstruction
confocal imaging
cytoskeleton
mechanical load
sclera
Journal
Journal of biophotonics
ISSN: 1864-0648
Titre abrégé: J Biophotonics
Pays: Germany
ID NLM: 101318567
Informations de publication
Date de publication:
01 2021
01 2021
Historique:
received:
26
05
2020
revised:
28
09
2020
accepted:
07
10
2020
pubmed:
11
10
2020
medline:
13
7
2021
entrez:
10
10
2020
Statut:
ppublish
Résumé
Computational models of cellular structures generally rely on simplifying approximations and assumptions that limit biological accuracy. This study presents a comprehensive image processing pipeline for creating unified three-dimensional (3D) reconstructions of the cell cytoskeletal networks and nuclei. Confocal image stacks of these cellular structures were reconstructed to 3D isosurfaces (Imaris), then tessellations were simplified to reduce the number of elements in initial meshes by applying quadric edge collapse decimation with preserved topology boundaries (MeshLab). Geometries were remeshed to ensure uniformity (Instant Meshes) and the resulting 3D meshes exported (ABAQUS) for downstream application. The protocol has been applied successfully to fibroblast cytoskeletal reorganisation in the scleral connective tissue of the eye, under mechanical load that mimics internal eye pressure. While the method herein is specifically employed to reconstruct immunofluorescent confocal imaging data, it is also more widely applicable to other biological imaging modalities where accurate 3D cell structures are required.
Identifiants
pubmed: 33038053
doi: 10.1002/jbio.202000202
doi:
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e202000202Informations de copyright
© 2020 The Authors. Journal of Biophotonics published by Wiley-VCH GmbH.
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