High-Throughput 3D Imaging Flow Cytometry of Suspended Adherent 3D Cell Cultures

Area of Science:

• Biomedical Engineering
• Cell Biology
• Microfluidics

Background:

• 3D cell cultures mimic in vivo environments, crucial for biological research.
• Hydrogel beads support adherent cells in spheroids, maintaining viability and function.
• Existing 3D imaging methods lack high-throughput and scalability for spheroid analysis.

Purpose of the Study:

• To develop a high-throughput, scalable 3D imaging flow cytometry platform for spheroid models.
• To enable precise quantification and comparison of cellular and subcellular structures in 3D cultures.
• To facilitate advanced biomedical analyses, including drug screening using spheroids or organoids.

Main Methods:

• Integration of single-objective fluorescence light-sheet microscopy with a microfluidic device.
• Utilizing hydrodynamic and acoustofluidic focusing techniques for cell manipulation.
• Development of an optofluidic platform for high-throughput 3D imaging of spheroids.

Main Results:

• The platform achieved unprecedented high-throughput and scalability, processing 1310 spheroids (28,117 cells/min).
• Enabled precise quantification of nuclear morphology in adhering versus suspended cells.
• Revealed that adhering cells exhibit smaller, less rounded nuclei compared to suspended cells.

Conclusions:

• The developed platform offers high throughput, robustness, and precision for 3D culture model analysis.
• It holds significant potential for image-based phenotypic screening and drug discovery.
• This technology advances the analysis of subcellular structures in complex 3D cell models.