Label-free light-sheet microfluidic cytometry for the automatic identification of senescent cells

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.

Identification of senescent cells in multipotent mesenchymal stromal cell cultures: Current methods and future directions

“…More advanced optical techniques have also been reported to be capable of identifying senescent cells. These include light-sheet [125,126] and digital holographic cytometry [127], methods which probe the biophysical aspects of cells to determine senescence. In particular, light-sheet cytometry uses the different scattering patterns of cells to determine their respective sizes and in turn senescence levels.…”

Applications of Light-Sheet Microscopy in Microdevices

“…A more elaborate setup shown previously (Miura et al, 2018) made use of flow-controlled conditions to perform the scan itself, removing the need of ETL or motorized stages. Similarly, glass capillaries have been used in flow cytometry for classification of phytoplankton (Wu J. et al, 2013) and identification of senescent cells (Lin et al, 2018). These devices hydrodynamically focus the cells in a square flow capillary.…”

“…Also, the use of LSFM in flow cytometry provides additional morphological information which is essential to some applications. This has allowed image processing and machine learning algorithms to perform automatic classification between different types of human cells (Lin et al, 2018) with potential applications in clinical diagnosis. In these studies, the square shape is important to avoid distortion of the light-sheet when it penetrates perpendicularly into the capillary.…”

Label‐free discrimination and selection of cancer cells from blood during flow using holography‐induced dielectrophoresis

“…However, in the past few years, there have been many advances in the development of label-free IFC for the analysis of cellular populations based on individual cell images. This includes the analysis of cancer cells (15)(16)(17), blood cells (18)(19)(20)(21)(22), cell cycle (23), cell differentiation (24) and drug response (25).…”