High-precision, low-complexity, high-resolution microscopy-based cell sorting.

Area of Science:

• Biomedical Engineering
• Cell Biology
• Microfluidics

Background:

• Image-activated cell sorting (IACS) leverages cellular features for isolating specialized cells.
• Current IACS methods face limitations in image quality and experimental complexity, hindering widespread adoption.

Purpose of the Study:

• To develop a simplified, high-performance IACS system.
• To improve image resolution and processing time for enhanced cell sorting accuracy.
• To demonstrate the system's capability in isolating viable T cells based on subcellular localization.

Main Methods:

• A novel microfluidic device integrating high numerical aperture wide-field microscopy.
• Precise dielectrophoretic cell manipulation for controlled handling.
• Advanced image analysis allowing extended processing times (hundreds of milliseconds).

Main Results:

• Achieved unprecedented image resolution of 216 nm in IACS.
• Successfully sorted live T cells based on subcellular fluorescence signals with >80% purity.
• Demonstrated high cell recovery rate (85%) and confirmed cell vitality post-sorting.

Conclusions:

• The developed microfluidic approach offers a low-complexity, high-resolution solution for IACS.
• This system overcomes previous limitations, making IACS a more accessible and powerful tool.
• The method ensures high purity, yield, and vitality of sorted cells, advancing cell isolation technologies.