Highly efficient combination of multiple single cells using a deterministic single-cell combinatorial reactor

Area of Science:

• Biotechnology
• Microfluidics
• Cell Biology

Background:

• Compartmentalization of single cells is vital for applications like single-cell transcriptome analysis and studying cell-cell interactions.
• High efficiency in pairing or combining single cells is essential to prevent analysis errors and misinterpretations.

Purpose of the Study:

• To develop a novel microfluidic device for highly efficient combination of multiple single cells.
• To enable precise trapping and combinatorial manipulation of individual cells for advanced biological assays.

Main Methods:

• Introduction of a deterministic single-cell combinatorial reactor (DSCR) device integrated with a multilayer interconnect Si/SiO2 control circuit.
• Sequential introduction and trapping of distinct cell populations into designated trap-wells using localized electric fields generated by cell-sized wells.
• Demonstration using PC3 cells stained with different fluorescent dyes, sequentially trapped in the DSCR.

Main Results:

• Achieved 93 ± 2% pairing efficiency for two cell populations.
• Demonstrated 82 ± 7% combination efficiency for three cell populations.
• Validated the device's capability for deterministic single-cell trapping and sequential operation.

Conclusions:

• The DSCR device offers a robust platform for the efficient combination of multiple single cells.
• This technology has significant potential for applications in analyzing multiple cell-cell communications and combinatorial indexing.
• The system provides a reliable method for precise manipulation of single cells in microfluidic environments.