3D Quantification of Vascular-Like Structures in z Stack Confocal Images

Ulrich Bonda; Anna Jaeschke; Anthony Lighterness; Jeremy Baldwin; Carsten Werner; Elena M De-Juan-Pardo; Laura J Bray

doi:10.1016/j.xpro.2020.100180

3D Quantification of Vascular-Like Structures in z Stack Confocal Images

STAR Protoc. 2020 Nov 21;1(3):100180. doi: 10.1016/j.xpro.2020.100180. eCollection 2020 Dec 18.

Authors

Ulrich Bonda¹, Anna Jaeschke^{2

3}, Anthony Lighterness², Jeremy Baldwin², Carsten Werner¹, Elena M De-Juan-Pardo^{3

4

5}, Laura J Bray^{2

3

6}

Affiliations

¹ Leibniz Institute of Polymer Research Dresden, Saxony 01069, Germany.
² Institute of Health and Biomedical Innovation, Queensland University of Technology (QUT), Brisbane, QLD 4059, Australia.
³ School of Mechanical, Medical and Process Engineering, Queensland University of Technology (QUT), Brisbane, QLD 4059, Australia.
⁴ Translational 3D Printing Laboratory for Advanced Tissue Engineering (T3mPLATE), Harry Perkins Institute of Medical Research, QEII Medical Centre, Nedlands and Centre for Medical Research, The University of Western Australia (UWA), Perth, WA 6009, Australia.
⁵ School of Engineering, The University of Western Australia, Perth, WA 6009, Australia.
⁶ ARC Training Centre for Cell and Tissue Engineering Technologies, Queensland University of Technology (QUT), Brisbane, QLD 4059, Australia.

Abstract

Optical slice microscopy is commonly used to characterize the morphometric features of 3D cellular cultures, such as in vitro vascularization. However, the quantitative analysis of those structures is often performed on a single 2D maximum intensity projection image, limiting the accuracy of data obtained from 3D cultures. Here, we present a protocol for the quantitative analysis of z stack images, utilizing Fiji, Amira, and WinFiber3D. This protocol facilitates the in-depth examination of vascular-like structures within 3D cell culture models. For complete details on the use and execution of this protocol, please refer to Koch et al. (2020).

Keywords: Cell culture; Microscopy; Tissue Engineering.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Algorithms
Blood Vessels / diagnostic imaging*
Imaging, Three-Dimensional*
Microscopy, Confocal / methods*
Staining and Labeling