Quantitative Vascular Structure Analysis

The samples were stained and analysed according to established protocols (Kniebs et al., 2019 (link); Kreimendahl et al., 2019 (link)). The cells were stained with the fluorescent Vybrant™ dyes DiO, Dil and DiD (ThermoFisher Scientific) according to manufacturer´s instructions. Additionally, the samples to determine the influence of tumor-specific therapeutics were stained with mouse anti-CD31 (PECAM-1, 1:100; Sigma-Aldrich) and Alexa Fluor^® 594 goat anti-mouse IgG (1:400, Acris Antibodies). The quantitative evaluation was performed with a two-photon laser scanning microscope, images processed and evaluated with Imaris 9.6.0 software (Bitplane Inc. South Winsor). The vascular structures were quantified according to their average volume, surface area, length, total number of vascular structures and branching points.

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Kniebs C., Luengen A.E., Guenther D., Cornelissen C.G., Schmitz-Rode T., Jockenhoevel S, & Thiebes A.L. (2021). Establishment of a Pre-vascularized 3D Lung Cancer Model in Fibrin Gel—Influence of Hypoxia and Cancer-Specific Therapeutics. Frontiers in Bioengineering and Biotechnology, 9, 761846.

Publication 2021

PECAM-1

Alexa 594 Anti igg Antibodies Cells Fluorescent dyes Goat Laser scanning microscope Mouse Pecam 1 Therapeutics Tumor Vascular structures

Corresponding Organization : RWTH Aachen University

Other organizations : Universitätsklinikum Aachen

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Variable analysis

independent variables

Tumor-specific therapeutics

dependent variables

Average volume of vascular structures
Surface area of vascular structures
Length of vascular structures
Total number of vascular structures
Number of branching points

control variables

Established protocols for staining and analysis (Kniebs et al., 2019; Kreimendahl et al., 2019)
Fluorescent Vybrant™ dyes DiO, Dil and DiD
Mouse anti-CD31 (PECAM-1, 1:100; Sigma-Aldrich)
Alexa Fluor® 594 goat anti-mouse IgG (1:400, Acris Antibodies)
Two-photon laser scanning microscope
Imaris 9.6.0 software (Bitplane Inc. South Winsor) for image processing and evaluation

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

Annotations

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