Multi-Zone Visco-Node-Pore Sensing: A Microfluidic Platform for Multi-Frequency Viscoelastic Phenotyping of Single...

Area of Science:

• Biophysics
• Cell Biology
• Microfluidics

Background:

• Single-cell analysis is crucial for understanding cellular heterogeneity.
• Viscoelastic properties are key indicators of cell state and function.
• Existing methods often lack the throughput or resolution for comprehensive phenotyping.

Purpose of the Study:

• To introduce multi-zone visco-Node-Pore Sensing (mz-visco-NPS), a novel microfluidic platform.
• To enable high-throughput, frequency-dependent viscoelastic phenotyping of individual cells.
• To investigate cellular heterogeneity across epithelial transformation states.

Main Methods:

• Development of an electronic-based microfluidic platform with sinusoidal contraction zones.
• Measurement of cell resistance changes at specific strain frequencies to determine elastic (G') and viscous (G″) moduli.
• Validation using MCF-7 cells with disrupted cytoskeletons.
• Application to diverse human mammary epithelial cell lines, including normal, immortalized, malignant, and metastatic cells.

Main Results:

• mz-visco-NPS accurately measures frequency-dependent viscoelastic moduli (G', G″).
• The platform successfully identified viscoelastic changes upon cytoskeletal disruption.
• Significant single-cell heterogeneity in viscoelastic properties was observed across epithelial cell continuum.
• High throughput (600 cells/hour) and ease-of-use were demonstrated.

Conclusions:

• mz-visco-NPS is an effective tool for single-cell viscoelastic phenotyping.
• The platform uncovers cellular heterogeneity masked by ensemble averaging.
• This technology has potential applications in cancer research and diagnostics by characterizing cell states.