New Method Enables Examination of Individual Immune Cells
Researchers at Karolinska Institutet and SciLifeLab in Sweden have developed a new method that enables examination of individual immune cells in blood samples, allowing scientists to distinguish between healthy and diseased cells. The technique, called spectral biophysical cytometry (SBC), measures multiple physical properties of individual cells simultaneously across very large cell populations, potentially advancing medical diagnostics and personalized treatment approaches.
A Breakthrough in Cellular Analysis
The physical properties of immune cells—such as how the cell membrane is structured or how mitochondria function—affect how cells move, communicate, and respond to inflammation. These properties have previously been difficult to measure on a large scale. The new method combines fluorescent nanosensors, which respond to the cells’ physical environment, with advanced spectral flow cytometry, where thousands of cells are analyzed one by one using laser light in a few minutes.
According to the study published in Nature Nanotechnology, the method measures three key biophysical properties simultaneously: membrane order, mitochondrial potential, and plasma membrane potential. This multiparametric approach provides an integrated picture of each cell’s health status at single-cell resolution.
Testing on Atherosclerosis Patients
The researchers tested the method on blood samples from 38 patients with atherosclerosis and 26 healthy control individuals. The analyses revealed clear differences between the two groups in cell membrane organization and mitochondrial function, particularly in different types of T cells—critical mediators in atherosclerotic disease progression.
“We show that immune cells in atherosclerosis have altered physical properties that can be measured directly in blood samples, without first needing to analyse genes or proteins,” said Erdinc Sezgin, researcher at SciLifeLab and the Department of Women’s and Children’s Health at Karolinska Institutet. “The method provides an integrated picture of the health status of the immune cells and can serve as a complement to more time-consuming and costly analyses.”
Sezgin, who leads the CSI:Nano Lab at Karolinska Institutet and SciLifeLab, added: “This suggests that the physical properties of cells are indications of health and important in disease processes in atherosclerosis and possibly also in other diseases.”
How the Method Works
The SBC platform uses three fluorescent nanosensors: Pro12A (reports membrane order), JC-1 (reports mitochondrial depolarization), and CV1 (reports membrane potential). Unlike traditional flow cytometry, which identifies cell types through selected surface proteins, SBC adds a functional layer by measuring the physical condition of each cell. The platform can process millions of cells while preserving single-cell resolution and uses commercially available instrumentation, making it potentially accessible to many laboratories.
The study also integrated biophysical measurements with lipidomics and transcriptomics to provide mechanistic insight into immune dysfunction in atherosclerosis. T-cell subsets showed significant alterations in membrane order and mitochondrial depolarization, reflecting coordinated changes in lipid composition and metabolic pathways.
Implications for Medical Diagnostics
The ability to distinguish atherosclerotic patients from healthy individuals based on the physical properties of immune cells suggests potential for biomarker discovery and disease monitoring. The method could potentially lead to faster, cheaper screening methods for immune-related diseases and enable monitoring of individual patients’ immune cell health status.
As reported by Xinhua News, researchers say this method can comprehensively reflect the health status of immune cells and can serve as a complement to time-consuming and costly analytical methods such as gene or protein analysis. The research was also covered by Mirage News.
Limitations and Next Steps
The study is a proof-of-concept demonstration, meaning an early step showing that the principle works rather than a validated clinical test. The researchers note that the cohort is not powered for subject-level diagnostic modeling, and the method requires further validation before clinical application.
The team plans to expand patient sample size and test the method in other diseases. Sezgin, who received the Biophysical Society’s Early Independent Career Award in 2025, emphasized that the physical properties of cells could play a role in disease processes beyond atherosclerosis.
“The method provides an integrated picture of the health status of the immune cells,” Sezgin said, “and can serve as a complement to more time-consuming and costly analyses.”
The research was funded by the Swedish Research Council, the Swedish Cancer Society, Wellcome Leap, and Karolinska Institutet. The researchers report no conflicts of interest.
What to Watch For
As the team expands its patient cohorts and explores applications in other diseases, the scientific community will be watching to see whether spectral biophysical cytometry can deliver on its promise of scalable, affordable immune cell health assessment. If validated, the method could transform how clinicians monitor immune function and potentially open new avenues for personalized medicine.