Monitoring Dynamic Immune Cell Responses Through Live-Cell Imaging 

A Celloger® Pro User Story from MNElab, Hanyang University

Understanding immune cell behavior requires more than endpoint analysis—it requires observing how cells respond and change over time.

Led by Professor Sung-Young Choi, the Micro & Nano Engineering Laboratory (MNElab) at Hanyang University develops advanced single-cell analysis and diagnostic technologies by integrating bioimaging, microfluidics, molecular engineering, optics, and machine learning.

The team’s current research focuses on microfluidic technologies for isolating CAR-T cells and neutrophils, as well as immune cell diagnostic technologies using NETosis and phagocytosis assays. To investigate these dynamic immune cell responses, the researchers use the Celloger® Pro to continuously monitor cellular behavior and fluorescence signals directly inside the incubator.

We spoke with one of the researchers to learn how the Celloger® Pro supports the lab’s immune cell research and which features have proven most useful in practice.

Observing Cells Without Disrupting the Culture Environment

Studying dynamic immune cell responses requires continuous observation to capture how cellular behavior changes over time under consistent culture conditions.

"We needed a way to observe changes in cell morphology and fluorescence signals over time while maintaining stable culture conditions. While searching for a suitable instrument, our professor recommended the Celloger® to us."

The Celloger® Pro is designed for real-time cell monitoring inside the incubator without disturbing the culture environment. During imaging, the camera moves across multiple positions while the culture vessel remains in place, supporting continuous observation under stable culture conditions.

Monitoring Fluorescence Signals in NETosis and Phagocytosis Assays

The team uses the Celloger® Pro’s dual fluorescence channels in NETosis and phagocytosis assays. By capturing signals from living cells and assay-specific targets simultaneously, the system allows the team to examine the relationship between cellular status and functional responses over time.

"With two fluorescence channels, we can follow changes in both the cells and target signals over time, giving us a more complete view of the response."

NETosis is a process in which neutrophils, a type of immune cell, release web-like structures made of DNA and proteins. These structures, known as neutrophil extracellular traps (NETs), help trap pathogens such as bacteria and limit their spread.

In the NETosis assay, SYTOX Green stains extracellular DNA, including DNA within NETs, as well as DNA in cells with compromised plasma membranes. CellTrace Red labels the cells for tracking, while bright-field imaging reveals changes in cell morphology. By combining these fluorescence signals with bright-field images, live-cell imaging allows the team to follow the cellular changes and DNA release associated with NET formation over time.

Phagocytosis is a process in which immune cells engulf particles, such as bacteria or cellular debris, and break down the internalized material. This helps clear pathogens and unwanted material.

In the phagocytosis assay, E. coli particles labeled with a pH-sensitive dye show increased green fluorescence as they reach acidic intracellular compartments following uptake, while SYTO 64 labels the cells in red for tracking. Monitoring both signals over time allows the team to track the cells and assess particle uptake through changes in green fluorescence.

Ease of Use in Everyday Research

Beyond its imaging capabilities, the team highlighted the system's compact design and intuitive software as practical benefits in daily research.

"The instrument is compact, so it doesn't take up much space in the incubator. The software is also intuitive, making it easy to use even for beginners.”

Its small footprint allows the Celloger® Pro to fit easily into the incubator while leaving enough space for other culture vessels. The software interface is clear and easy to navigate, allowing first-time users to quickly understand the key functions and set up imaging experiments with minimal training.

Sharper Imaging with Z-Stacking

The team also pointed to Z-stacking as a particularly useful feature in their research.

"The Celloger® Pro’s Z-stacking captures images at multiple focal planes along the Z-axis, allowing us to detect and analyze signals at different focal depths more accurately."

To learn more about the Celloger® Pro and its features, visit the product page. [Celloger® Pro product page]

Toward a Deeper Understanding of Immune Cell Responses

Building on this work, MNElab will continue to investigate immune cell responses and translate the insights gained into the further development of its diagnostic technologies.

We thank the MNElab team for sharing their research and their experience using the Celloger® Pro.