METHODS article
Front. Bioeng. Biotechnol.
Sec. Organoids and Organ-On-A-Chip
Volume 13 - 2025 | doi: 10.3389/fbioe.2025.1585003
This article is part of the Research TopicAdvancements in Image Processing and Analysis Techniques for Microphysiological SystemsView all 3 articles
Methods for Processing and Analyzing Images of Vascularized Micro-Organ and Tumor Systems
Provisionally accepted
Stephanie J. Hachey1*
Christopher Hatch1
Daniela Gaebler1
Alexander G Forsythe2
Makena Ewald1
Alexander L Chopra1
Jennifer Fang3
Chris Hughes1- 1University of California, Irvine, Irvine, United States
- 2Simon Fraser University, Burnaby, British Columbia, Canada
- 3Tulane University, New Orleans, Louisiana, United States
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Our group has developed and validated an advanced microfluidic platform to improve preclinical modeling of healthy and disease states, enabling extended culture and detailed analysis of tissueengineered miniaturized organ constructs, or "organs-on-chips." Within this system, diverse cell types self-organize into perfused microvascular networks under dynamic flow within tissue chambers, effectively mimicking the structure and function of native tissues. This setup facilitates physiological intravascular delivery of nutrients, immune cells, and therapeutic agents, and creates a realistic microenvironment to study cellular interactions and tissue responses. Known as the vascularized micro-organ (VMO), this adaptable platform can be customized to represent various organ systems or tumors, forming a vascularized micro-tumor (VMT) for cancer studies. The VMO/VMT system closely simulates in vivo nutrient exchange and drug delivery within a 3D microenvironment, establishing a high-fidelity model for drug screening and mechanistic studies in vascular biology, cancer, and organ-specific pathologies.Furthermore, the optical transparency of the device supports high-resolution, real-time imaging of fluorescently labeled cells and molecules within the tissue construct, providing key insights into drug responses, cell interactions, and dynamic processes such as epithelial-mesenchymal transition. To manage the extensive imaging data generated, we created standardized, high-throughput workflows for image analysis. This manuscript presents our image processing and analysis pipeline, utilizing a suite of tools in Fiji/ImageJ to streamline data extraction from the VMO/VMT model, substantially reducing manual processing time. Additionally, we demonstrate how these tools can be adapted for analyzing imaging data from traditional in vitro models and microphysiological systems developed by other researchers.
Keywords: microphysiological system, Tumor Microenvironment, Tumor on chip, microfluidic, Bioengineering, image processing, Therapeutic development
Received: 28 Feb 2025; Accepted: 13 May 2025.
Copyright: © 2025 Hachey, Hatch, Gaebler, Forsythe, Ewald, Chopra, Fang and Hughes. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Stephanie J. Hachey, University of California, Irvine, Irvine, United States
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