High throughput single cell separation and identification using a self-priming isometric and Equant screw valve-based...

Area of Science:

• Biotechnology
• Microfluidics
• Cell Biology

Background:

• Single cell research is advancing rapidly due to new separation and identification platforms.
• Microfluidic chip-based strategies are crucial for single cell analysis.
• Efficient high-throughput methods are needed for isolating and identifying single cells.

Purpose of the Study:

• To develop a novel microfluidic chip for high-throughput single cell isolation and identification.
• To demonstrate the chip's capability in analyzing specific gene expression in cancer cells.
• To assess the potential of the chip for resource-limited and point-of-care settings.

Main Methods:

• A self-priming isometric and Equant screw valve-based microfluidic chip (SIES chip) was designed and fabricated.
• Special features include a peripheral water tank for pressure balance and screw valves for sample loading.
• Multistage branching "T" shape channels ensure even cell distribution for analysis of up to 2000 cells.
• The chip was used to isolate and identify single A549 cells expressing the activated leukocyte cell adhesion molecule (ALCAM) gene.

Main Results:

• The SIES chip successfully isolated and identified single A549 cells.
• Analysis revealed that approximately 5.1% of A549 cells expressed the ALCAM gene.
• This finding could aid in the reclassification of A549 cells.

Conclusions:

• The developed SIES chip offers an inexpensive, user-friendly, and portable solution for high-throughput single cell analysis.
• The technology shows promise for applications in resource-limited environments and point-of-care diagnostics.
• The chip's ability to identify specific gene expression in single cells has implications for cell reclassification and further research.