Patient-derived tumoroids are widely used in comparative oncology. These translationally relevant models preserve the source tissue’s clinically meaningful features while offering scalable, patient-specific 3D systems suitable for diagnostic discovery, mechanistic study, and therapeutic screening.
Image Credit: Yokogawa Fluid Imaging Technologies, Inc.
These models are especially valuable when sample availability is limited, because 3D culture expansion can yield enough material for repeated downstream studies and further characterization.
Intrinsic heterogeneity across and within tumoroid subtypes necessitates analytical methods that capture representative population-level data, because low-throughput visual inspection is rarely adequate for this task.
However, capturing this data is difficult because tumoroid growth is complex. Cultivating tumoroids in vitro typically requires scaffold-based culture matrices to support tissue-like spatial patterning, organization, and contact-mediated signaling. Extracellular matrix components can adversely affect direct visualization of tumoroid responses in vitro.
Some assays can be performed with tumoroids embedded in scaffold matrices, but evaluating microscopic behavior effectively generally requires matrix removal to reduce imaging artifacts.
Matrix removal creates its own challenges, however, because extracting tumoroids from their supportive 3D microenvironment risks cell loss, disruption of tissue architecture, and morphological deformation. This compromises viability, representativeness, structural integrity, and reproducibility.
This article explores flow imaging microscopy (FIM) with FlowCam as a high-throughput, non-destructive method for characterizing complex 3D samples.
FlowCam was used to analyze low-passage canine tumoroids cultured in Matrigel®. This process quantified particle count, morphology, size, and structural complexity across hundreds of individual 3D structures in a single run.
The process proved highly consistent with the more labor-intensive, lower-throughput manual microscopy approach.
Digital image analysis of four patient-derived subtypes showed distinct size distributions and enabled semi-automated classification of sheet-like structures and heterogeneous clusters, supporting the refinement of culture workflows and the representative assessment of tumoroid heterogeneity.
These findings highlight FIM with FlowCam as a promising new means of analyzing early 3D cell culture responses where there is a need to both generate representative datasets and preserve heterogeneity.
Figure 1. Representative workflow for quantitative analysis of low-passage canine tumoroids cultured in Matrigel® using FlowCam and comparative brightfield microscopy. Image Credit: Yokogawa Fluid Imaging Technologies, Inc.
Read the full application note to explore the methodology and results in greater detail, including comparisons with manual brightfield microscopy, differences across four patient-derived tumoroid subtypes, morphology-based classification, and the potential of FlowCam for quality control, therapeutic screening, diagnostic discovery, and broader cell therapy workflows.
Read the Full App Note here
About Yokogawa Fluid Imaging Technologies, Inc.
At Yokogawa Fluid Imaging Technologies our mission is to bring clarity and focus to the study of subvisible particles in the life sciences. With our FlowCam technology, we are committed to developing innovative hardware and software to monitor Earth’s water bodies and drinking water supply for hazardous algae, to help make vaccines and injectable medicines safer, and to improve product quality in a variety of applications.
FlowCam instruments combine the benefits of digital imaging, flow cytometry, and microscopy into a single solution - Flow Imaging Microscopy (FIM). Beyond traditional particle sizing and counting, FIM's image-based analysis allows for comprehensive characterization of subvisible API aggregates and contaminants in biopharmaceuticals, mammalian cells, microplankton, emulsions, and advanced materials. FlowCam instruments have been deployed in over 50 countries, supporting research, development, and environmental monitoring in the life sciences, materials research, and industrial applications.
In the video below you will learn about flow imaging microscopy and how FlowCam works to provide count, size, characterization, and digital images of subvisible particles.
FlowCam Explainer Video - What is Flow Imaging Microscopy?
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