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Gel microdroplets and flow cytometry: rapid determination of antibody secretion by individual cells within a cell
1Harvard-M.I.T. Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge 02139.
Insights
This study introduces a novel method using flow cytometry and gel microdroplets (GMDs) to rapidly measure macromolecule secretion from individual cells. The technique efficiently identifies antibody-secreting cell subpopulations within hours.
Area of Science:
- Biotechnology
- Cell Biology
- Immunology
Background:
- Measuring macromolecule secretion from individual cells in large populations is crucial for biological research.
- Existing methods can be time-consuming and lack single-cell resolution for secreted products.
Purpose of the Study:
- To develop and validate a rapid, high-throughput method for quantifying macromolecule secretion at the single-cell level.
- To identify and characterize subpopulations of secreting cells within a heterogeneous population.
Main Methods:
- Integration of flow cytometry with gel microdroplets (GMDs) for single-cell encapsulation.
- Utilized agarose microparticles (53-88 μm diameter) for cell containment, ensuring one cell per GMD.
- Employed solid-phase fluorescence immunoassay within GMDs to detect captured secreted molecules.
Main Results:
- Successfully determined the secretion of macromolecules from individual cells within a large population.
- Demonstrated the method's efficacy using a model system of antibody-secreting hybridoma and non-secreting masticytoma cells.
- Identified antibody-secreting cell subpopulations in under fifteen hours.
Conclusions:
- The combined flow cytometry and GMD approach offers a rapid and efficient means to analyze single-cell secretion.
- This method significantly reduces the time required to identify functional cell subpopulations.
- The technique holds potential for applications in antibody discovery, cell line characterization, and therapeutic development.
Abstract:
We report a new method capable of rapidly determining the secretion of biologically important macromolecules from each of many individual cells within a large population. This method combines flow cytometry with gel microdroplets (GMDs), which in this study were agarose particles ranging from about 53 to 88 mu in diameter. The GMDs were formed from a liquid 2.5% agarose suspension with cells at a concentration which yielded mostly zero or one cell per GMD. A large number of extracellular binding sites were also provided within each GMD, allowing the capture of secreted molecules, and their subsequent measurement by solid phase, fluorescence immunoassay. The method was explored using a model system of mouse hybridoma (secreting) and mouse masticytoma (non-secreting) cells. The method was able to determine subpopulations of individual cells that secreted antibody in less than fifteen hours after receipt of a conventional cell suspension.