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Analyzing Cellular Internalization of Nanoparticles and Bacteria by Multi-spectral Imaging Flow Cytometry

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Published on: June 8, 2012

Intelligent image-activated sorting of large cells enabled by elasto-inertial focusing.

Yuzuki Nagasaka¹, Akihiro Isozaki^1,2, Hiroki Matsumura^1,3

¹Department of Chemistry, Graduate School of Science, The University of Tokyo, Tokyo 113-0033, Japan. tding@g.ecc.u-tokyo.ac.jp.

|February 25, 2026

Summary

This study introduces a new image-activated cell sorting (IACS) method using elasto-inertial focusing to sort large cells and objects at high speeds. This breakthrough overcomes limitations of previous systems, enabling efficient sorting of complex biological samples.

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Area of Science:

Biotechnology
Microfluidics
Cell Sorting

Background:

Image-activated cell sorting (IACS) is crucial for linking cell morphology and function at the single-cell level.
Existing IACS methods struggle with sorting large cells and objects due to focusing instability in microfluidic devices.
Hydrodynamic focusing limits the speed and purity of IACS for larger biological entities.

Purpose of the Study:

To develop an IACS system capable of sorting large cells and objects at high speeds.
To overcome the limitations of hydrodynamic focusing in conventional IACS systems.
To enable high-content, real-time image analysis for sorting large biological entities.

Main Methods:

Developed an IACS system integrating elasto-inertial focusing for stable particle manipulation.
Validated the elasto-inertial focuser with large particles (>20 μm) over extended distances (≥35 mm).
Utilized a convolutional neural network classifier for sorting size-mixed particles and microalgae (Euglena gracilis).

Main Results:

Demonstrated stable focusing of large particles (>20 μm) using elasto-inertial focusing.
Achieved 96.0% purity and 80.5% yield for size-mixed particle sorting at 172 events per second.
Successfully sorted Euglena gracilis based on lipid droplet formation, achieving 4.5-fold enrichment at 128 events per second.

Conclusions:

Elasto-inertial focusing enables high-speed IACS for large cells and objects, overcoming previous limitations.
The developed IACS system maintains high sorting purity, yield, and event rates for complex biological samples.
This technology advances single-cell analysis and sorting capabilities for scientific and industrial applications involving large cellular entities.