High-throughput multimodal optofluidic biophysical imaging cytometry.

Area of Science:

• Biophysics
• Cell Biology
• Analytical Chemistry

Background:

• Traditional biophysical cytometry offers limited cellular phenotyping, hindering the study of cellular heterogeneity.
• Existing methods often rely on a few biophysical readouts, restricting comprehensive analysis.
• There is a need for high-throughput platforms capable of detailed single-cell biophysical characterization.

Purpose of the Study:

• To introduce a novel multimodal biophysical cytometry platform, quantitative phase morpho-rheological (QP-MORE) cytometry.
• To enable simultaneous capture of high-resolution biophysical and mechanical phenotypes of single cells at ultrahigh throughput.
• To demonstrate QP-MORE's capability in dissecting drug-induced cellular heterogeneity and classifying drug mechanisms.

Main Methods:

• QP-MORE cytometry integrates ultrafast single-cell quantitative phase imaging (QPI) and high-throughput deformability cytometry.
• Utilizes a microfluidic constriction channel design for label-free, multi-contrast imaging of cells.
• Employs a robust calibration protocol for accurate morpho-rheological measurements.

Main Results:

• QP-MORE achieves subcellular resolution and captures whole-cell rheology in a single pass.
• The platform revealed distinct, drug-specific subcellular biophysical signatures in leukemia and breast cancer cells.
• QP-MORE achieved 99% accuracy in classifying drug mechanisms, outperforming traditional deformability cytometry (78-94%).

Conclusions:

• QP-MORE cytometry significantly expands the capabilities of biophysical analysis for single cells.
• This platform offers unprecedented insights into cellular heterogeneity and drug interactions.
• QP-MORE holds great potential for advancing cell biology research and drug discovery.