High accuracy particle analysis using sheathless microfluidic impedance cytometry

Spencer, Daniel; Caselli, Federica; Bisegna, Paolo; Morgan, Hywel

doi:10.1039/c6lc00339g

Cited by 76 publications

(78 citation statements)

References 32 publications

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“…In this fluidic situation and within the used electrode configuration the system detects two preferred signal amplitudes, a smaller amplitude for the cells in the middle of the channel, and a larger amplitude for cells on the top or bottom of the channel, which are closer to the electrodes. The slight phase shift leading to the J-shape is an effect of electronic cross-talk between the electrode pairs [49]. This cross-talk is substantial at low frequencies (0.5 MHz), but almost inexistent at high frequencies (12 MHz).…”

Section: Resultsmentioning

confidence: 99%

Impedance Flow Cytometry: A Novel Technique in Pollen Analysis

Heidmann

Schade-Kampmann²,

Lambalk

et al. 2016

PLoS ONE

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IntroductionAn efficient and reliable method to estimate plant cell viability, especially of pollen, is important for plant breeding research and plant production processes. Pollen quality is determined by classical methods, like staining techniques or in vitro pollen germination, each having disadvantages with respect to reliability, analysis speed, and species dependency. Analysing single cells based on their dielectric properties by impedance flow cytometry (IFC) has developed into a common method for cellular characterisation in microbiology and medicine during the last decade. The aim of this study is to demonstrate the potential of IFC in plant cell analysis with the focus on pollen.MethodDeveloping and mature pollen grains were analysed during their passage through a microfluidic chip to which radio frequencies of 0.5 to 12 MHz were applied. The acquired data provided information about the developmental stage, viability, and germination capacity. The biological relevance of the acquired IFC data was confirmed by classical staining methods, inactivation controls, as well as pollen germination assays.ResultsDifferent stages of developing pollen, dead, viable and germinating pollen populations could be detected and quantified by IFC. Pollen viability analysis by classical FDA staining showed a high correlation with IFC data. In parallel, pollen with active germination potential could be discriminated from the dead and the viable but non-germinating population.ConclusionThe presented data demonstrate that IFC is an efficient, label-free, reliable and non-destructive technique to analyse pollen quality in a species-independent manner.

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Section: Resultsmentioning

confidence: 99%

Impedance Flow Cytometry: A Novel Technique in Pollen Analysis

Heidmann

Schade-Kampmann²,

Lambalk

et al. 2016

PLoS ONE

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“…Resulting calibration parameters (Eq. 5) position determination could be replicated in a vertical configuration using five pairs of facing electrodes (Spencer et al 2016), thus providing an even more accurate estimate of particle vertical position, at the expense of a more complex microfabrication process. The density plot of the electrical velocity V against the optical velocity V opt is shown in Fig.…”

Section: Optical Validation Of Electrical Position (X Y)mentioning

confidence: 99%

“…The second signal, denoted I OBQ , is the difference in electric current flowing through two sensing electrodes with a z-coordinate different from the stimulating electrodes (the 1st and the 5th electrodes along z) featuring a current path oblique with respect to particle flow. An analogous wiring scheme was introduced in Spencer et al (2016) for a chip comprising five pairs of facing electrodes, aimed at solving the positional dependence issue. and an oblique measurement ( I OBQ ).…”

Section: Electrical Estimate Of Particle Lateral Positionmentioning

confidence: 99%

“…Event detection in the data streams was performed with a previously reported algorithm , and a simple MATLAB script was used for event feature extraction by template fitting (Spencer et al 2016;De Ninno et al 2017). In particular, for each detected event the ratio R (Eq.…”

Section: Impedance Data Acquisition and Processingmentioning

confidence: 99%

“…along channel width) (Caselli et al , 2018 or vertical (i.e. along channel height) (Spencer et al 2016;De Ninno et al 2017) particle position.…”

Section: Introductionmentioning

confidence: 99%

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Electrical measurement of cross-sectional position of particles flowing through a microchannel

Reale

Ninno

Businaro

et al. 2018

Microfluid Nanofluid

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The first high-throughput system for the electrical detection of cross-sectional position and velocity of individual particles flowing through a rectangular microchannel is presented. Lateral position (along channel width) and vertical position (along channel height) are measured using two different sets of coplanar electrodes. In particular, the ratio of travel times measured with electrodes generating a current flow transverse or oblique with respect to particle trajectory yields lateral position. The relative prominence and transit time of a bipolar double-Gaussian signal obtained with a suited electrode configuration, respectively, supply vertical position and velocity. The operating principle is presented by means of finite element numerical simulations. The method is experimentally validated by comparing the electrical estimates of position and velocity of polystyrene beads with optical estimates obtained by processing high-speed images. The system is used to observe bead focusing at different particle Reynolds numbers. This system, providing a fully electrical characterization of single-particle motion, represents a powerful tool, e.g. to understand fluid motion at the microscale, in particle separation studies, or to assess the performance of particle focusing devices. Moreover, it can be simultaneously used to perform single-cell impedance spectroscopy, thus achieving an unprecedented multiparamteric characterization.

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Supervised machine learning in microfluidic impedance flow cytometry for improved particle size determination

et al. 2022

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The assessment of particle and cell size in electrical microfluidic flow cytometers has become common practice. Nevertheless, in flow cytometers with coplanar electrodes accurate determination of particle size is difficult, owing to the inhomogeneous electric field. Pre-defined signal templates and compensation methods have been introduced to correct for this positional dependence, but are cumbersome when dealing with irregular signal shapes. We introduce a simple and accurate post-processing method without the use of pre-defined signal templates and compensation functions using supervised machine learning. We implemented a multiple linear regression model and show an average reduction of the particle diameter variation by 37% with respect to an earlier processing method based on a feature extraction algorithm and compensation function. Furthermore, we demonstrate its application in flow cytometry by determining the size distribution of a population of small (4.6 ± 0.9 μm) and large (5.9 ± 0.8 μm) yeast cells. The improved performance of this coplanar, two electrode chip enables precise cell size determination in easy to fabricate impedance flow cytometers.

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High accuracy particle analysis using sheathless microfluidic impedance cytometry

Cited by 76 publications

References 32 publications

Impedance Flow Cytometry: A Novel Technique in Pollen Analysis

Impedance Flow Cytometry: A Novel Technique in Pollen Analysis

Electrical measurement of cross-sectional position of particles flowing through a microchannel

Supervised machine learning in microfluidic impedance flow cytometry for improved particle size determination

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