Detecting cells rotations for increasing the robustness of cell sizing by impedance measurements, with or without machine learning.

The Coulter principle is a widespread technique for sizing red blood cells (RBCs) in hematological analyzers. It is based on the monitoring of the electrical perturbations generated by cells passing through a micro-orifice, in which a concentrated electrical field is imposed by two electrodes. However, artifacts associated with near-wall passages in the sensing region are known to skew the statistics for RBCs sizing. This study presents numerical results that emphasize the link between the cell flow-induced rotation in the detection area and the error in its measured volume. Based on these observations, two methods are developed to identify and reject pulses impaired by cell rotation. In the first strategy, the filtering is allowed by a metric computed directly from the waveform. In the second, a numerical database is employed to train a neural network capable of detecting if the cell has experienced a rotation, given its electrical pulse. Detecting and rejecting rotation-associated pulses are shown to provide results comparable to hydrodynamical focusing, which enforces cells to flow in the center of the orifice, the gold standard implementation of the Coulter principle.

Red blood cell rheology during a complete blood count: A proof of concept.

Counting and sizing blood cells in hematological analyzers is achieved using the Coulter principle. The cells flow in a micro-aperture in which a strong electrical field is imposed, so that an electrical perturbation, called pulse, is measured each time a cell crosses the orifice. The pulses are expected to contain information on the shape and deformability of Red Blood Cells (RBCs), since recent studies state that RBCs rotate and deform in the micro-orifice. By implementing a dedicated numerical model, the present study sheds light on a variety of cells dynamics, which leads to different associated pulse signatures. Furthermore, simulations provide new insights on how RBCs shapes and mechanical properties affect the measured signals. Those numerical observations are confirmed by experimental assays. Finally, specific features are introduced for assessing the most relevant characteristics from the various pulse signatures and shown to highlight RBCs alterations induced by drugs. In summary, this study paves the way to a characterization of RBC rheology by routine hematological instruments.

New erythrocyte parameters derived from the Coulter principle relate with red blood cell properties-A pilot study in diabetes mellitus.

In routine hematological instruments, blood cells are counted and sized by monitoring the impedance signals induced during their passage through a Coulter orifice. However, only signals associated with centered paths in the aperture are considered for analysis, while the rejected measurements, caused by near-wall trajectories, can provide additional information on red blood cells (RBC), as recent publications suggest. To assess usefulness of two new parameters in describing alterations in RBC properties, we performed a pilot study to compare blood samples from patients with diabetes mellitus (DM), frequent pathological condition associated with impairment in RBC deformability, versus controls. A total of 345 blood samples were analyzed: 225 in the DM group and 120 in the control group. A diagram of [Formula: see text] and [Formula: see text], the two new parameters derived from the analysis of impedancemetry pulses, was used to compare distribution of RBC subpopulations between groups. To discriminate RBC from DM and control individuals, based on our multiparametric analysis, we built a score from variables derived from [Formula: see text] matrix which showed good performances: area under the receiving operating characteristic curve 0.948 (0.920-0.970), p<0.0001; best discriminating value: negative predictive value 94.7%, positive predictive value was 78.4%. These results seem promising to approach RBC alterations in routine laboratory practice. The related potential clinically relevant outcomes remain to be investigated.

Detection of circulating microfilariae in canine EDTA blood using lens-free technology: preliminary results.

Dirofilaria immitis causes life-threatening heart disease in dogs, thus screening of dog populations is important. Lens-free technology (LFT) is a low-cost imaging technique based on light diffraction that allows computerized recognition of small objects in holographic images. We evaluated an algorithm capable of recognizing microfilariae in canine whole blood using the LFT. We examined 3 groups of 10 EDTA blood specimens, from dogs with microfilaremia (group A), healthy dogs (B), and dogs with hematologic modifications other than microfilaremia (C). The LFT analyzer photographed repeated series of 5 images of all samples. The algorithm declared a sample positive if a microfilaria was detected on ≥1, ≥2, or ≥3 of the 5 images of a series. Microfilariae were detected visually in the images in 9 of 10 cases in group A; no microfilariae were seen in the images from groups B and C. Of the 30 cases, there were 14, 4, and only 3 false-positives with the 1 of 5, 2 of 5, and 3 of 5 image cutoffs, respectively. There were no false-negatives, regardless of cutoff. LFT seems useful for detecting microfilaria and could have application in clinical pathology.

Numerical simulation of deformable particles in a Coulter counter.

In Coulter counters, cells counting and volumetry are achieved by monitoring their electrical print when they flow through a sensing zone. However, the volume measurement may be impaired by the cell dynamics, which may be difficult to control. In this paper, numerical simulations of the dynamics and electrical signature of red blood cells in a Coulter counter are presented, accounting for the deformability of the cells. In particular, a specific numerical pipeline is developed to overcome the challenge of the multi-scale nature of the problem. It consists in segmenting the whole computation of the cell dynamics and electrical response in a series of dedicated computations, with a saving of one order of magnitude in computational time. This numerical pipeline is used with rigid spheres and deformable red blood cells in an industrial Coulter counter geometry, and compared with experimental measurements. The simulations not only reproduce electrical signatures typical of those measured experimentally, but also allow an analysis of the electrical signature in terms of the heterogeneity of the electrical field and dynamics of the particles in the measurement zone. This study provides a methodology for computing the sizing of rigid or deformable particles by Coulter counters, opening the way to a better understanding of cells signatures in such devices.

Numerical simulation of particle dynamics in an orifice-electrode system. Application to counting and sizing by impedance measurement.

This paper describes how to numerically tackle the problem of counting and sizing particles by impedance measurement in an orifice-electrode system. The model allows to simulate the particle dynamics submitted to strong hydrodynamic stresses through a microorifice and to compute the voltage pulses generated by the modification of the inner dielectric medium. This approach gives important information about particles size distribution and allows to quantify the role of trajectory and orientation of particles on the size measurement.