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Overview Of Cell Separation And Isolation01:20

Overview Of Cell Separation And Isolation

Cell separation was first achieved in 1964 by S. H. Seal, who separated large tumor cells from the smaller blood cells using filtration. Two years later, Pohl and Hawk performed experiments on how cells respond differently to a nonuniform electric field based on the cell type. Such observations were the inception of cell separation methods, which allow isolating a single cell type from a heterogeneous sample.

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Microfluidics-based High-throughput Circulating Tumor Cell Sorting and Single-cell Sequencing Technology
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Single-cell analysis and sorting using droplet-based microfluidics.

Linas Mazutis1, John Gilbert, W Lloyd Ung

  • 1School of Engineering and Applied Sciences (SEAS), Harvard University, Cambridge, Massachusetts, USA.

Nature Protocols
|April 6, 2013
PubMed
Summary
This summary is machine-generated.

This study introduces droplet microfluidics for rapid single-cell analysis and sorting. The method efficiently detects secreted antibodies from hybridoma cells, enabling high-throughput screening.

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Area of Science:

  • Biotechnology
  • Microfluidics
  • Cell Biology

Background:

  • Traditional methods like flow cytometry have limitations in analyzing secreted cellular products.
  • Single-cell analysis requires efficient methods for high-throughput screening.

Purpose of the Study:

  • To develop a droplet-based microfluidics protocol for high-throughput analysis and sorting of single cells.
  • To demonstrate a binding assay for detecting antibodies secreted from single mouse hybridoma cells.

Main Methods:

  • Single cells were compartmentalized in 50-pl droplets with fluorescent probes and antibody-coated beads.
  • Secreted antibodies were captured by beads, and binding was detected via fluorescence localization.
  • Droplet sorting was performed at approximately 200 Hz.

Main Results:

  • Secreted antibodies were detected within 15 minutes.
  • The system enabled efficient droplet sorting and cell enrichment.
  • Screening approximately 1 million cells took 2-6 hours of microfluidic operation.

Conclusions:

  • Droplet microfluidics offers a powerful platform for analyzing secreted proteins from single cells.
  • This approach overcomes limitations of traditional flow cytometry and cell sorting.
  • The system is adaptable for screening various proteins and cellular activities.