Functional Phenotype Flow Cytometry: On Chip Sorting of Individual Cells According to Responses to Stimuli

Area of Science:

• Biotechnology
• Cell Biology
• Microfluidics

Background:

• Cellular heterogeneity is key to understanding biological systems.
• Effective cell separation is vital for studying cell subpopulations.

Purpose of the Study:

• To develop a novel device, the functional phenotype flow cytometer (FPFC), for cell separation.
• To enable cell sorting based on real-time phenotypic responses to stimuli.

Main Methods:

• A microfluidic platform introduces cells to a stimulus reagent.
• Cells are incubated and sorted based on intracellular fluorescence readouts.
• The FPFC processes cells at a throughput of up to 4 cells/min.

Main Results:

• The FPFC successfully sorts cells based on their phenotypic responses.
• Intracellular fluorescence was used as the primary readout method.
• Hundreds of cells were profiled and sorted within hours.

Conclusions:

• The functional phenotype flow cytometer (FPFC) offers a new method for cell subpopulation isolation.
• This technology aids in the study of cellular heterogeneity and responses.
• The FPFC was validated using human Burkitt's lymphoma cells and B cell antigen receptor activation.