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  • Wiley  (12)
  • Chen, Jian  (12)
  • 1
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    Inherent bioelectrical parameters of hundreds of thousands of single leukocytes based on impedance flow cytometry
    Wiley ; 2022
    In:  Cytometry Part A Vol. 101, No. 8 ( 2022-08), p. 630-638
    Details
    In: Cytometry Part A, Wiley, Vol. 101, No. 8 ( 2022-08), p. 630-638
    Abstract: As label‐free biomarkers, bioelectrical properties of single cells have been widely used in hematology analyzers for 3‐part differential of leukocytes, in which, however, instrument dependent bioelectrical parameters (e.g., DC/AC impedance values) rather than inherent bioelectrical parameters (e.g., diameter D c , specific membrane capacitance C sm and cytoplasmic conductivity σ cy ) were used, leading to poor comparisons among different instruments. In order to address this issue, this study collected inherent bioelectrical parameters from hundreds of thousands of white blood cells based on a home‐developed impedance flow cytometry with corresponding 3‐part differential of leukocytes realized. More specifically, leukocytes were separated into three major subtypes of granulocytes, monocytes and lymphocytes based on density gradient centrifugation. Then these separated cells were aspirated through a constriction‐microchannel based impedance flow cytometry where inherent bioelectrical parameters of D c , C sm and σ cy were quantified as 9.8 ± 0.7 μm, 2.06 ± 0.26 μF/cm 2 , and 0.34 ± 0.05 S/m for granulocytes (n cell  = 134,829); 10.4 ± 1.0 μm, 2.45 ± 0.48 μF/cm 2 , and 0.42 ± 0.08 S/m for monocytes (n cell  = 40,226); 8.0 ± 0.5 μm, 2.23 ± 0.34 μF/cm 2 , and 0.35 ± 0.08 S/m for lymphocytes (n cell  = 129,193). Based on these inherent bioelectrical parameters, neural pattern recognition was conducted, producing a high “classification accuracy” of 93.5% in classifying these three subtypes of leukocytes. These results indicate that as inherent bioelectrical parameters, D c , C sm , and σ cy can be used to electrically phenotype white blood cells in a label‐free manner.
    Type of Medium: Online Resource
    ISSN: 1552-4922 , 1552-4930
    URL: Issue
    URL: Article
    DOI: 10.1002/cyto.v101.8
    DOI: 10.1002/cyto.a.24544
    Language: English
    Publisher: Wiley
    Publication Date: 2022
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  • 2
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    Microfluidic Cytometry for High‐Throughput Characterization of Single Cell Cytoplasmic Viscosity Using Crossing Constriction Channels
    Wiley ; 2020
    In:  Cytometry Part A Vol. 97, No. 6 ( 2020-06), p. 630-637
    Details
    In: Cytometry Part A, Wiley, Vol. 97, No. 6 ( 2020-06), p. 630-637
    Abstract: This article presents an approach of microfluidic flow cytometry capable of continuously characterizing cytoplasmic viscosities of single cells. The microfluidic system consists of a major constriction channel and a side constriction channel perpendicularly crossing each other. Cells are forced to rapidly travel through the major channel and are partially aspirated into the side channel when passing the channel junction. Numerical simulations were conducted to model the time dependence of the aspiration length into the side channel, which enables the measurement of cytoplasmic viscosity by fitting the model results to experimental data. As a demonstration for high‐throughput measurement, the cytoplasmic viscosities of HL‐60 cells that were native or treated by N ‐Formylmethionine‐leucyl‐phenylalanine (fMLP) were quantified with sample sizes as large as thousands of cells. Both the average and median cytoplasmic viscosities of native HL‐60 cells were found to be about 10% smaller than those of fMLP‐treated HL‐60 cells, consistent with previous observations that fMLP treatment can increase the rigidity of white blood cells. Furthermore, the microfluidic system was used to process granulocytes from three donors (sample size 〉 1,000 cells for each donor). The results revealed that the cytoplasmic viscosity of granulocytes from one donor was significantly higher than the other two, which may result from the fact that this donor just recovered from an inflammation. In summary, the developed microfluidic system can collect cytoplasmic viscosities from thousands of cells and may function as an enabling tool in the field of single‐cell analysis. © 2019 International Society for Advancement of Cytometry
    Type of Medium: Online Resource
    ISSN: 1552-4922 , 1552-4930
    URL: Issue
    URL: Article
    DOI: 10.1002/cyto.a.v97.6
    DOI: 10.1002/cyto.a.23921
    Language: English
    Publisher: Wiley
    Publication Date: 2020
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  • 3
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    Mechanical property characterization of hundreds of single nuclei based on microfluidic constriction channel
    Wiley ; 2018
    In:  Cytometry Part A Vol. 93, No. 8 ( 2018-08), p. 822-828
    Details
    In: Cytometry Part A, Wiley, Vol. 93, No. 8 ( 2018-08), p. 822-828
    Abstract: As label‐free biomarkers, the mechanical properties of nuclei are widely treated as promising biomechanical markers for cell type classification and cellular status evaluation. However, previously reported mechanical parameters were derived from only around 10 nuclei, lacking statistical significances due to low sample numbers. To address this issue, nuclei were first isolated from SW620 and A549 cells, respectively, using a chemical treatment method. This was followed by aspirating them through two types of microfluidic constriction channels for mechanical property characterization. In this study, hundreds of nuclei were characterized, producing passage times of 0.5 ± 1.2 s for SW620 nuclei in type I constriction channel ( n  = 153), 0.045 ± 0.047 s for SW620 nuclei in type II constriction channel ( n  = 215) and 0.50 ± 0.86 s for A549 nuclei in type II constriction channel. In addition, neural network based pattern recognition was used to classify the nuclei isolated from SW620 and A549 cells, producing successful classification rates of 87.2% for diameters of nuclei, 85.5% for passage times of nuclei and 89.3% for both passage times and diameters of nuclei. These results indicate that the characterization of the mechanical properties of nuclei may contribute to the classification of different tumor cells.
    Type of Medium: Online Resource
    ISSN: 1552-4922 , 1552-4930
    URL: Issue
    URL: Article
    DOI: 10.1002/cyto.v93.8
    DOI: 10.1002/cyto.a.23386
    Language: English
    Publisher: Wiley
    Publication Date: 2018
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  • 4
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    Classification of tumor subtypes leveraging constriction‐channel based impedance flow cytometry and optical imaging
    Wiley ; 2021
    In:  Cytometry Part A Vol. 99, No. 11 ( 2021-11), p. 1114-1122
    Details
    In: Cytometry Part A, Wiley, Vol. 99, No. 11 ( 2021-11), p. 1114-1122
    Abstract: As label‐free biomarkers, electrical properties of single cells have been widely used for cell‐type classification and cell‐status evaluation. However, as intrinsic bioelectrical markers, previously reported membrane capacitance and cytoplasmic resistance (e.g., specific membrane capacitance C specific membrane and cytoplasmic conductivity σ cytoplasm ) of tumor subtypes were derived from tens of single cells, lacking statistical significance due to low cell numbers. In this study, tumor subtypes were constructed based on phenotype (treatment with 4‐methylumbelliferone) or genotype (knockdown of ROCK1) modifications and then aspirated through a constriction‐channel based impedance flow cytometry to characterize single‐cell C specific membrane and σ cytoplasm . Thousands of single tumor cells with phenotype modifications were measured, resulting in significant differences in 1.64 ± 0.43 μF/cm 2 vs. 1.55 ± 0.47 μF/cm 2 of C specific membrane and 0.96 ± 0.37 S/m vs. 1.24 ± 0.47 S/m of σ cytoplasm for 95C cells (792 cells of 95C‐control vs. 1529 cells of 95C‐pheno‐mod); 2.56 ± 0.88 μF/cm 2 vs. 2.33 ± 0.56 μF/cm 2 of C specific membrane and 0.83 ± 0.18 S/m vs. 0.93 ± 0.25 S/m of σ cytoplasm for H1299 cells (962 cells of H1299‐control vs. 637 cells of H1299‐pheno‐mod). Furthermore, thousands of single tumor cells with genotype modifications were measured, resulting in significant differences in 3.82 ± 0.92 vs. 3.18 ± 0.47 μF/cm 2 of C specific membrane and 0.47 ± 0.05 vs. 0.52 ± 0.05 S/m of σ cytoplasm (1100 cells of A549‐control vs. 1100 cells of A549‐geno‐mod). These results indicate that as intrinsic bioelectrical markers, specific membrane capacitance and cytoplasmic conductivity can be used to classify tumor subtypes.
    Type of Medium: Online Resource
    ISSN: 1552-4922 , 1552-4930
    URL: Issue
    URL: Article
    DOI: 10.1002/cyto.a.v99.11
    DOI: 10.1002/cyto.a.24358
    Language: English
    Publisher: Wiley
    Publication Date: 2021
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  • 5
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    Toward five‐part differential of leukocytes based on electrical impedances of single cells and neural network
    Wiley ; 2023
    In:  Cytometry Part A Vol. 103, No. 5 ( 2023-05), p. 439-446
    Details
    In: Cytometry Part A, Wiley, Vol. 103, No. 5 ( 2023-05), p. 439-446
    Abstract: The five‐part differential of leukocytes plays key roles in the diagnosis of a variety of diseases and is realized by optical examinations of single cells, which is prone to various artifacts due to chemical treatments. The classification of leukocytes based on electrical impedances without cell treatments has not been demonstrated because of limitations in approaches of impedance acquisition and data processing. In this study, based on treatment‐free single‐cell impedance profiles collected from impedance flow cytometry leveraging constriction microchannels, two types of neural pattern recognition were conducted for comparisons with the purpose of realizing the five‐part differential of leukocytes. In the first approach, 30 features from impedance profiles were defined manually and extracted automatically, and then a feedforward neural network was conducted, producing a classification accuracy of 84.9% in the five‐part leukocyte differential. In the second approach, a customized recurrent neural network was developed to process impedance profiles directly and based on deep learning, a classification accuracy of 97.5% in the five‐part leukocyte differential was reported. These results validated the feasibility of the five‐part leukocyte differential based on label‐free impedance profiles of single cells and thus provide a new perspective of differentiating white blood cells based on impedance flow cytometry.
    Type of Medium: Online Resource
    ISSN: 1552-4922 , 1552-4930
    URL: Issue
    URL: Article
    DOI: 10.1002/cyto.a.v103.5
    DOI: 10.1002/cyto.a.24697
    Language: English
    Publisher: Wiley
    Publication Date: 2023
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  • 6
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    Advance of microfluidic constriction channel system of measuring single‐cell cortical tension/specific capacitance of membrane and conductivity of cytoplasm
    Wiley ; 2022
    In:  Cytometry Part A Vol. 101, No. 5 ( 2022-05), p. 434-447
    Details
    In: Cytometry Part A, Wiley, Vol. 101, No. 5 ( 2022-05), p. 434-447
    Abstract: This paper reported a microfluidic platform which realized the characterization of inherent single‐cell biomechanical and bioelectrical parameters simultaneously. Individual cells traveled through a constriction channel with deformation images and impedance variations captured and processed into cortical tension T c , specific membrane capacitance C sm , and cytoplasmic conductivity σ cy based on an equivalent biophysical model. These properties of thousands of individual cells of K562, Jurkat, HL‐60, HL‐60 treated with paraformaldehyde (PA)/cytochalasin D (CD)/concanavalin A (ConA), granulocytes of Donor 1, Donor 2, and Donor 3 were quantified for the first time. Leveraging T c , C sm , and σ cy , (1) high accuracies of classifying wild‐type and processed HL‐60 cells (e.g., 93.5% of PA treated vs. CD treated HL‐60 cells) were realized, revealing the effectiveness of using these three biophysical parameters in cell‐type classification; (2) low accuracies of classifying normal granulocytes from three donors (e.g., 56.4% of Donor 1 vs. 2), indicating comparable parameters for normal granulocytes. In conclusion, this platform can characterize single‐cell T c , C sm , and σ cy concurrently and quantify multiple parameters in single‐cell analysis.
    Type of Medium: Online Resource
    ISSN: 1552-4922 , 1552-4930
    URL: Issue
    URL: Article
    DOI: 10.1002/cyto.a.v101.5
    DOI: 10.1002/cyto.a.24517
    Language: English
    Publisher: Wiley
    Publication Date: 2022
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  • 7
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    Inherent single‐cell bioelectrical parameters of thousands of neutrophils, eosinophils and basophils derived from impedance flow cytometry
    Wiley ; 2022
    In:  Cytometry Part A Vol. 101, No. 8 ( 2022-08), p. 639-647
    Details
    In: Cytometry Part A, Wiley, Vol. 101, No. 8 ( 2022-08), p. 639-647
    Abstract: Single‐cell bioelectrical properties are commonly used for blood cell phenotyping in a label‐free manner. However, previously reported inherent single‐cell bioelectrical parameters (e.g., diameter D c , specific membrane capacitance C sm and cytoplasmic conductivity σ cy ) of neutrophils, eosinophils and basophils were obtained from only tens of individual cells with limited statistical significance. In this study, granulocytes were separated into neutrophils, eosinophils and basophils based on fluorescent flow cytometry, which were further aspirated through a constriction‐microchannel impedance flow cytometry for electrical property characterization. Based on this microfluidic impedance flow cytometry, single‐cell values of D c , C sm and σ cy were measured as 10.25 ± 0.66 μm, 2.17 ± 0.30 μF/cm 2 , and 0.37 ± 0.05 S/m for neutrophils ( n cell  = 9442); 9.73 ± 0.51 μm, 2.07 ± 0.19 μF/cm 2 , and 0.30 ± 0.04 S/m for eosinophils ( n cell  = 2982); 9.75 ± 0.49 μm, 2.06 ± 0.17 μF/cm 2 , and 0.31 ± 0.04 S/m for basophils ( n cell  = 5377). Based on these inherent single‐cell bioelectrical parameters, neural pattern recognition was conducted, producing classification rates of 80.8% (neutrophil vs. eosinophil), 77.7% (neutrophil vs. basophil) and 59.3% (neutrophil vs. basophil). These results indicate that as inherent single‐cell bioelectrical parameters, D c , C sm and σ cy can be used to classify neutrophils from eosinophils or basophils to some extent while they cannot be used to effectively distinguish eosinophils from basophils.
    Type of Medium: Online Resource
    ISSN: 1552-4922 , 1552-4930
    URL: Issue
    URL: Article
    DOI: 10.1002/cyto.v101.8
    DOI: 10.1002/cyto.a.24559
    Language: English
    Publisher: Wiley
    Publication Date: 2022
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  • 8
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    Leukocyte differential based on an imaging and impedance flow cytometry of microfluidics coupled with deep neural networks
    Wiley ; 2024
    In:  Cytometry Part A
    Details
    In: Cytometry Part A, Wiley
    Abstract: The differential of leukocytes functions as the first indicator in clinical examinations. However, microscopic examinations suffered from key limitations of low throughputs in classifying leukocytes while commercially available hematology analyzers failed to provide quantitative accuracies in leukocyte differentials. A home‐developed imaging and impedance flow cytometry of microfluidics was used to capture fluorescent images and impedance variations of single cells traveling through constrictional microchannels. Convolutional and recurrent neural networks were adopted for data processing and feature extractions, which were then fused by a support vector machine to realize the four‐part differential of leukocytes. The classification accuracies of the four‐part leukocyte differential were quantified as 95.4% based on fluorescent images plus the convolutional neural network, 90.3% based on impedance variations plus the recurrent neural network, and 99.3% on the basis of fluorescent images, impedance variations, and deep neural networks. Based on single‐cell fluorescent imaging and impedance variations coupled with deep neural networks, the four‐part leukocyte differential can be realized with almost 100% accuracy.
    Type of Medium: Online Resource
    ISSN: 1552-4922 , 1552-4930
    URL: Article
    DOI: 10.1002/cyto.a.24823
    Language: English
    Publisher: Wiley
    Publication Date: 2024
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  • 9
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    Clinical anatomy of superior laryngeal artery via transoral approach
    Wiley ; 2022
    In:  Laryngoscope Investigative Otolaryngology Vol. 7, No. 3 ( 2022-06), p. 702-706
    Details
    In: Laryngoscope Investigative Otolaryngology, Wiley, Vol. 7, No. 3 ( 2022-06), p. 702-706
    Abstract: Hemorrhage is the most common complication caused by transoral laryngopharyngeal surgery. It is believed that proper management of the superior laryngeal artery (SLA), the main feeding artery for the larynx and pharynx, may reduce intra‐ and postoperative hemorrhage incidence. The aim of this study was to illustrate the anatomy of the SLA via transoral endoscopic approach. Methods Fourteen sides of SLA from heads of seven fresh‐frozen and silicone‐injected cadavers were dissected. Transoral dissections were performed for the intra‐laryngeal segment of SLA, and transcervical dissections were performed to confirm the anatomical measurements. Results SLA had a slightly descending course from the origin to the larynx, and there was a major branch supplying the epiglottis, named pharyngo‐epiglottic artery (PEA). Parallel with the internal superior laryngeal nerve (ISLN), SLA passed through the thyrohyoid membrane and ended into the hypopharynx. The distance from SLA to the superior horn of thyroid cartilage (SHTC) was (9.11 ± 0.58)mm on the left and (9.01 ± 0.37)mm on the right; the distance from SLA to the inferior margin of the hyoid bone (IMHB) was (2.00 ± 0.11)mm on the left and (1.95 ± 0.08)mm on the right; the distance from SLA to ISLN was (5.98 ± 0.48)mm on the left and (5.78 ± 0.36)mm on the right. No significant difference was found between bilateral sides ( p   〉  0.05). Moreover, the distance from SLA to superior margin of thyroid cartilage (SMTC) was (5.52 ± 0.24)mm on the left and (5.80 ± 0.15)mm on the right. A significant difference was also found between bilateral sides ( p  = 0.03), which might suggest the SLA is located further from the SMTC on the right side. Conclusion SHTC, SMTC, and IMHB could be regarded as anatomical landmarks to locate SLA when applying a transoral approach. Moreover, a complete understanding of the detailed anatomy of the superior laryngeal artery may improve the detection of hemostasis in transoral laryngeal or hypo‐pharyngeal surgery.
    Type of Medium: Online Resource
    ISSN: 2378-8038 , 2378-8038
    URL: Issue
    URL: Article
    DOI: 10.1002/lio2.v7.3
    DOI: 10.1002/lio2.781
    Language: English
    Publisher: Wiley
    Publication Date: 2022
    detail.hit.zdb_id: 2851702-7
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  • 10
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    A microfabricated 96‐well wound‐healing assay
    Wiley ; 2017
    In:  Cytometry Part A Vol. 91, No. 12 ( 2017-12), p. 1192-1199
    Details
    In: Cytometry Part A, Wiley, Vol. 91, No. 12 ( 2017-12), p. 1192-1199
    Abstract: This article presents a microfabricated 96‐well wound‐healing assay enabling high‐throughput measurement of cellular migration capabilities. Within each well, the middle area is the wound region, made of microfabricated gold surface with self‐assembled PEG repellent for cell seeding. After the formation of a cellular confluent monolayer around the wound region, collagen solution was applied to form three‐dimensional matrix to cover the PEG surface, initiating the wound‐healing process. By interpreting the numbers of migrated cells into the wound regions as a function of specific stimuli with different concentrations, EC50 (half‐maximal effective concentration) was obtained. Using H1299 as a model, values of EC50 were quantified as 8% and 160 ng/ml for fetal bovine serum and CXCL12, respectively. In addition, the values of EC50 were demonstrated not to be affected by variations in compositions of extracellular matrix and geometries of wounds, which can thus be regarded as an intrinsic marker. Furthermore, the migration capabilities of a second cell type (HeLa) were characterized by the developed wound‐healing assay, producing EC50 of 2% when fetal bovine serum was used as the stimuli. These results validated the proposed high‐throughput wound‐healing assay, which may function as an enabling tool in studying cellular capabilities of migration and invasion. © 2017 International Society for Advancement of Cytometry
    Type of Medium: Online Resource
    ISSN: 1552-4922 , 1552-4930
    URL: Issue
    URL: Article
    DOI: 10.1002/cyto.a.v91.12
    DOI: 10.1002/cyto.a.23286
    Language: English
    Publisher: Wiley
    Publication Date: 2017
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