# Label‐Free Leukocyte Biophysical Profiling Using Impedance‐Deformability Cytometry for Rapid Cardiovascular Risk Stratification

## TL;DR

A new label-free method profiles neutrophil biophysical traits to assess cardiovascular risk in type 2 diabetes patients.

## Contribution

Development of a high-throughput, label-free microfluidic platform for rapid cardiovascular risk stratification in T2DM using neutrophil biophysical profiling.

## Key findings

- DM-CVD neutrophils show distinct impedance signatures and pro-inflammatory transcriptomic profiles.
- Principal component analysis accurately identifies vascular dysfunction in T2DM patients.
- Impedance-deformability cytometry reveals biophysical changes linked to inflammation and hyperglycemia.

## Abstract

Type 2 diabetes mellitus (T2DM) presents a global health burden with cardiovascular disease (CVD) as its leading cause of mortality. A rapid clinical‐adaptable microfluidic workflow for label‐free CVD risk profiling based on neutrophil biophysical abnormalities is developed. This high‐throughput single cell (>1,000 cells min−1) “electro‐mechano‐phenotyping” method integrates Uniform Manifold Approximation and Projection analysis to assess leukocyte biophysical changes (size, deformability, and impedance properties) linked to inflammation, thrombosis, and hyperglycemia in vitro, and in diabetic and diabetic atherosclerosis‐prone mouse models. In a clinical study of healthy, pre‐diabetes, diabetes, and diabetic patients with CVD (DM‐CVD) subjects (n = 10–11 per group), DM‐CVD neutrophils exhibited a distinct impedance signature and pro‐inflammatory transcriptomic profile marked by cytoskeletal dysregulation and altered RhoA signaling. Principal component analysis (area under the curve = 0.971) identifies individuals with vascular dysfunction exhibiting increased carotid intima‐media thickness and reduced reactive hyperemia index. These findings support impedance‐based neutrophil profiling as a promising, cost‐effective strategy for cardiovascular risk stratification in T2DM.

This study presents an impedance‐based single‐cell profiling platform that quantifies the electrical and mechanical properties of neutrophils across in vitro, in vivo, and clinical samples. The approach reveals distinct biophysical alterations associated with type 2 diabetes (T2DM) and cardiovascular complications, suggesting its potential utility for functional, label‐free assessment of immune dysregulation and cardiovascular risk stratification in T2DM.