A high-throughput technique to map cell images to cell positions using a 3D imaging flow cytometer

Area of Science:

• Biotechnology
• Cell Biology
• Imaging Science

Background:

• Existing spatial biology methods are unsuitable for nonadherent cells.
• Accurate spatial mapping of single cells is crucial for understanding biological processes.

Purpose of the Study:

• To develop a high-throughput technique for relating individual nonadherent cell positions to their 3D imaging features.
• To enable single-cell resolution spatial mapping for cells not amenable to traditional tissue-based methods.

Main Methods:

• Utilized a custom 3D imaging flow cytometer (3D-IFC) for high-throughput 3D cell imaging (500-1,000 cells/s).
• Employed a robotic cell placement platform (CPP) for first-in-first-out (FIFO) cell dispensing.
• Developed a marker bead and DNA sequencing method to detect and prevent FIFO errors.

Main Results:

• Successfully mapped 3D scattering, fluorescent, and 2D transmission images of ~100,000 human cancer cells to their locations in under 10 minutes.
• Demonstrated the feasibility of high-throughput spatial mapping for nonadherent cells.
• Validated the accuracy of the cell placement and error detection system.

Conclusions:

• The developed technique provides a robust method for high-throughput spatial mapping of nonadherent cells.
• This technology bridges the gap between single-cell image analysis and single-cell molecular analysis.
• The methodology is adaptable to various imaging modalities beyond the custom 3D-IFC.