Abstract
Label-free microfluidic cytometry is of increasing interest for single cell analysis due to its advantages of high-throughput, miniaturization, as well as noninvasive detection. Here we develop a next generation label-free light-sheet microfluidic cytometer for single cell analysis by two-dimensional (2D) light scattering measurements. Our cytometer integrates light sheet illumination with a disposable hydrodynamic focusing unit, which can achieve 3D hydrodynamic focusing of a sample fluid to a diameter of 19 micrometer without microfabrication. This integration also improves the signal to noise ratio (SNR) for the acquisition of 2D light scattering patterns from label-free cells. Particle sizing with submicron resolution is achieved by our light-sheet flow cytometer, where Euclidean distance-based similarity measures are performed. Label-free, automatic classification of senescent and normal cells is achieved with a high accuracy rate by incorporating our light-sheet flow cytometry with support vector machine (SVM) algorithms. Our light-sheet microfluidic cytometry with a microfabrication-free hydrodynamic focusing unit may find wide applications for automatic and label-free clinical diagnosis.
Highlights
Identification of senescent cells is important for the better understanding of aging, cancer and many other age-related diseases [1, 2]
Our results demonstrate that the light-sheet microfluidic cytometry with a microfabrication-free hydrodynamic focusing unit may be used for label-free analysis of senescence-related disease
The hydrodynamic focusing unit we developed here is composed by two glass capillaries that are fixed by specially designed connectors
Summary
Identification of senescent cells is important for the better understanding of aging, cancer and many other age-related diseases [1, 2]. Cell senescence are characterized by histochemical or fluorescence-based detection of biomarkers [4, 5]. The activity of SA-β-gal (a widely used biomarker for senescent cells) can be measured by using flow cytometry with fluorescence reagents [6]. The fluorescence-based detection relies on specific biomarkers of cells to perform characterization, which is sophisticated and time-consuming. The wide adoption of flow cytometer for cell analysis is limited. There is a great need to develop label-free microfluidic device because of its high-throughput, miniaturization, as well as non-invasive detection for single-cell analysis
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