Multiplex single particle analysis in microfluidics

Dannhauser, David; Romeo, Giovanni; Causa, Filippo; Santo, Ilaria De; Netti, Paolo A.

doi:10.1039/c4an01033g

Cited by 29 publications

(29 citation statements)

References 18 publications

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“…The recorded scattering images were generated by the interaction of a narrow-collimated incident laser beam (wavelength of 632.8 nm) -passing a highly transparent microfluidic channel from the bottom- and a viscoelastic aligned individual particle or cell while flowing in our microfluidic system with submicron range resolution. The basic working principle and used optical equipment are reported elsewhere 47 in more detail.…”

Section: Methodsmentioning

confidence: 99%

Single-cell screening of multiple biophysical properties in leukemia diagnosis from peripheral blood by pure light scattering

Dannhauser

Rossi

Ripaldi

et al. 2017

Sci Rep

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Histology and histopathology are based on the morphometric observations of quiescent cells. Their diagnostic potential could largely benefit from a simultaneous screening of intrinsic biophysical properties at single-cell level. For such a purpose, we analyzed light scattering signatures of individual mononuclear blood cells in microfluidic flow. In particular, we extracted a set of biophysical properties including morphometric (dimension, shape and nucleus-to-cytosol ratio) and optical (optical density) ones to clearly discriminate different cell types and stages. By considering distinctive ranges of biophysical properties along with the obtained relative cell frequencies, we can identify unique cell classes corresponding to specific clinical conditions (p < 0.01). Based on such a straightforward approach, we are able to discriminate T-, B-lymphocytes, monocytes and beyond that first results on different stages of lymphoid and myeloid leukemia cells are presented. This work shows that the simultaneous screening of only three biophysical properties enables a clear distinction between pathological and physiological mononuclear blood stream cells. We believe our approach could represent a useful tool for a label-free analysis of biophysical single-cell signatures.

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Section: Methodsmentioning

confidence: 99%

Single-cell screening of multiple biophysical properties in leukemia diagnosis from peripheral blood by pure light scattering

Dannhauser

Rossi

Ripaldi

et al. 2017

Sci Rep

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“…More detailed information about the microfluidic device and general alignment conditions for rigid latex particles can be seen elsewhere 9 . An aspheric lens with a high numerical aperture collects the emitted forward scattering of each cell passing through the beam.…”

Section: Light Scatteringmentioning

confidence: 99%

Cells characterization in microfluidic flows by small angle light scattering and 3D holographic technique

et al. 2015

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The Light Scattering Profile (LSP) of an individual cell provides a fast and accurate characterization of its morphological properties. By combining a camera-based small angle light scattering apparatus with a microfluidic-induced particle migration technique, it is possible to characterize cells in microfluidic flows. The scattering profile of an individual cell can be fully characterized by our optimized optical light collection system. Viscoelastic-induced particle migration by polyethylene oxide implemented in a low-cost microfluidic device composed of an alignment section and a measuring section opens the possibility of precise, label-free, individual cell analysis. We have studied living cells in microfluidic flows by our light scattering apparatus and by a Digital Holographic Microscope (DHM) system. Our DHM measurements provided an accurate 3D position tracking even in multiple cell conditions.

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“…We used a scattering apparatuswhose design and basic principle are shown elsewhere 25 to accurately measure the LSPs of differently sized individual particles or cells in microfluidic flows. The obtained LSPs span from 2-30°with an optical resolution of 0.1022°, enabling erythrocyte diameter distinction of at least 0.1 μm, due to the effort of the implemented narrow incident laser beam collimation (λ = 632.8 nm) and optimized scattered light collection system.…”

Section: Light Scattering Setupmentioning

confidence: 99%

“…25 The average scattering intensity of each scattering angle was combined to a continuous LSP and characterized by matching the best fitting individual cell simulation. 25 The average scattering intensity of each scattering angle was combined to a continuous LSP and characterized by matching the best fitting individual cell simulation.…”

Section: Lsp Acquisition and Simulationmentioning

confidence: 99%

Optical signature of erythrocytes by light scattering in microfluidic flows

et al. 2015

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A camera-based light scattering approach coupled with a viscoelasticity-induced cell migration technique has been used to characterize the morphological properties of erythrocytes in microfluidic flows. We have obtained the light scattering profiles (LSPs) of individual living cells in microfluidic flows over a wide angular range and matched them with scattering simulations to characterize their morphological properties. The viscoelasticity-induced 3D cell alignment in microfluidic flows has been investigated by bright-field and holographic microscopy tracking, where the latter technique has been used to obtain precise cell alignment profiles in-flow. Such information allows variable cell probability control in microfluidic flows at very low viscoelastic polymer concentrations, obtaining cell measurements that are almost physiological. Our results confirm the possibility of precise, label-free analysis of individual living erythrocytes in microfluidic flows.

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Multiplex single particle analysis in microfluidics

Cited by 29 publications

References 18 publications

Single-cell screening of multiple biophysical properties in leukemia diagnosis from peripheral blood by pure light scattering

Single-cell screening of multiple biophysical properties in leukemia diagnosis from peripheral blood by pure light scattering

Cells characterization in microfluidic flows by small angle light scattering and 3D holographic technique

Optical signature of erythrocytes by light scattering in microfluidic flows

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