Light sheet based volume flow cytometry (VFC) for rapid volume reconstruction and parameter estimation on the go

Area of Science:

• Biomedical Optics
• Cellular Imaging
• Microfluidics

Background:

• Optical imaging is crucial for disease diagnosis and monitoring progression.
• Existing techniques face limitations in throughput and resolution for live specimens.

Purpose of the Study:

• To introduce and validate the Volumetric Flow Cytometry (VFC) imaging technique for enhanced live cell analysis.
• To demonstrate VFC's capability for high-throughput, organelle-level imaging and parameter estimation.

Main Methods:

• Developed a sheet-based VFC system integrated with a multichannel microfluidic chip.
• Calibrated the system using fluorescent beads to account for flow-induced aberrations.
• Employed computational deconvolution with flow-variant point spread functions for volume reconstruction.
• Analyzed mitochondrial networks in HeLa cancer cells in real-time.

Main Results:

• Achieved single-shot sectional visualization and instant volume reconstruction with organelle-level resolution.
• Enabled parallel visualization of multiple cells at high flow rates (up to 2000 nl/min).
• Successfully quantified real-time cellular parameters including mitochondria count, size, morphology, and cell strain.

Conclusions:

• The VFC system provides a powerful platform for high-throughput, real-time monitoring of sub-cellular organelle organization.
• Quantifiable cellular parameters derived from VFC imaging can reveal disease states and metastatic progression.
• VFC enhances biomedical imaging with high-content capacity for live specimens.