Clear 3D Cell Culture Models With Minimal Changes to Morphology and Without Compromising Sensitivity
Despite the growing relevance of 3D cell culture as a research model, imaging techniques used to characterize these models are highly limited. Due to the thickness and opacity of 3D cellular structures, most current imaging technologies cannot penetrate to the center of the tissues, resulting in only the outer 2 to 3 layers of cells being detected. This causes the dark centers often seen in images of 3D cell culture models, which proves highly problematic for accurate analysis as these outer cells are most exposed to compounds, nutrients and oxygen and thus do not reflect the entire cell population.
Corning now offers 3D Clear Tissue Clearing reagent that can be used in a tissue clearing technique, designed specifically to support imaging for 3D cell culture models and plate-based high throughput processing. When paired with fluorescent labeling (e.g., fluorescent protein, immunofluorescence, chemical dyes) and high content confocal microscopy, Corning 3D Clear reagent allows for complete 3D cell culture model characterization and more accurate drug screening.
Complete Model Characterization
In this figure it is demonstrated that tissue clearing combined with confocal microscopy allows for a 3-4 times increase in cells detected (C) when compared a 3D cell culture model cleared with Corning 3D Clear reagent (B) and a 3D cell culture model imaged in PBS without clearing (A).
Increased Dose Response Sensitivity
The use of Corning 3D Clear Tissue Clearing reagent allows for a significant increase in dose response sensitivity (left), as the technique allows for the entire population of cells within a 3D cell culture model to be characterized. Furthermore, due to interrogation of the entire cell population, assessment of drug effect is more accurate. Without clearing, only the exterior cells are surveyed, and as such, measurements of cell proliferation are overestimated, since the outer layers of cells show more proliferation than the interior cells.
Organoid Culture Models
Corning 3D Clear can be used for clearing organoid models to enable optimal clarity for sharp, clear cell images.
Disease models support our understanding of how disease develops and progresses, as well as potential treatment approaches.
3D multicellular spheroids can develop metabolic gradients that create heterogeneous cell populations with superior cell-to-cell and cell-to-ECM interactions.
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