Hydrodynamic stretching of single cells for large population mechanical phenotyping

TLDR

Cell state is commonly assessed via biochemical and biophysical markers, yet while biophysical deformability offers a label‑free advantage, existing methods for measuring it have limited clinical adoption. The study demonstrates an automated microfluidic platform that probes single‑cell deformability at approximately 2,000 cells per second. The platform employs inertial focusing to deliver cells into a high‑strain extensional flow, where high‑speed imaging and computational analysis extract quantitative deformability parameters. The system achieves throughput orders of magnitude higher than prior tools, accurately predicts disease state in cancer and immune‑activation patients with 91 % sensitivity and 86 % specificity, and identifies early stem‑cell differentiation biomarkers linked to nuclear changes, thereby enabling clinical diagnostics, stem‑cell characterization, and single‑cell biophysics.

Abstract

Cell state is often assayed through measurement of biochemical and biophysical markers. Although biochemical markers have been widely used, intrinsic biophysical markers, such as the ability to mechanically deform under a load, are advantageous in that they do not require costly labeling or sample preparation. However, current techniques that assay cell mechanical properties have had limited adoption in clinical and cell biology research applications. Here, we demonstrate an automated microfluidic technology capable of probing single-cell deformability at approximately 2,000 cells/s. The method uses inertial focusing to uniformly deliver cells to a stretching extensional flow where cells are deformed at high strain rates, imaged with a high-speed camera, and computationally analyzed to extract quantitative parameters. This approach allows us to analyze cells at throughputs orders of magnitude faster than previously reported biophysical flow cytometers and single-cell mechanics tools, while creating easily observable larger strains and limiting user time commitment and bias through automation. Using this approach we rapidly assay the deformability of native populations of leukocytes and malignant cells in pleural effusions and accurately predict disease state in patients with cancer and immune activation with a sensitivity of 91% and a specificity of 86%. As a tool for biological research, we show the deformability we measure is an early biomarker for pluripotent stem cell differentiation and is likely linked to nuclear structural changes. Microfluidic deformability cytometry brings the statistical accuracy of traditional flow cytometric techniques to label-free biophysical biomarkers, enabling applications in clinical diagnostics, stem cell characterization, and single-cell biophysics.