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Flow Cytometry Study of Immune Cell Subpopulations from theMouse Vertebral BoneMarrow and Intervertebral Disc Following Endplate Microfracture 认领 引用
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作者 Dalin Wang Mingcai Zhang +3 位作者 Richard Hastings Patrick George Ryan Ranzau Jinxi Wang 《BIOCELL》 SCIE 2026年第4期1-17,共17页
Objective:Although endplate(EP)injury may cause intervertebral disc(IVD)degeneration andModic changes(MCs)in the vertebral bone marrow(VBM),EP injury-induced synchronous cellular reactions and their crosstalk in the I... Objective:Although endplate(EP)injury may cause intervertebral disc(IVD)degeneration andModic changes(MCs)in the vertebral bone marrow(VBM),EP injury-induced synchronous cellular reactions and their crosstalk in the IVDandVBMremain unclear.This protocol-based study aimed to streamline and optimize themethods of tissue harvest and cell preparation for flow cytometry(FCM)analysis of T-cell and macrophage subpopulations in both VBM and IVD adjacent to the surgically induced EP microfracture in mice.Methods:EP injury or sham procedure was performed at the spinal levels L4-5 and L5-6 in male mice.Step-by-step techniques on the harvest of lumbar VBMand IVDtissues,isolation of tissue-specific cells,and generation of single cell suspensions were described.FCM analysis was performed using specific antibodies against cell-surface markers and multi-color cytometry for precise delineation of cell subsets to explore the cellular mechanism ofMCs.An extracellular staining assay to identify macrophage subsets,as well as extra-and intra-cellular staining assays to identify T lymphocyte subsets fromthe VBM and IVD,were performed and optimized.Results:FCM analysis demonstrated that significantly increased macrophage numbers and M2a polarization response were observed in VBM samples from the sham and EP injury groups,while the ratio of M2aotal number of macrophages was significantly increased and the ratio of M1/M2a was significantly decreased in IVDs from the sham and EP injury groups.A significantly increased Treg cell response was detected in VBM samples from the EP injury group,but not the sham group.Conclusions:This protocol reports novel and reproducible methods of tissue harvest,cell preparation,and antibody selection for flow cytometry analysis of T-cell and macrophage subpopulations isolated from the VBM and IVD following EP injury inmice.This protocolmay be utilized for exploring the cellular mechanism of MCs and IVD degeneration in animal models. 展开更多
关键词 Flow cytometry immune cell subsets bone marrow intervertebral disc cartilaginous endplate spine
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Holotomography-driven learning unlocks in-silico staining of single cells in flow cytometry by avoiding fluorescence co-registration 认领 引用
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作者 Daniele Pirone Giusy Giugliano +12 位作者 Michela Schiavo Annalaura Montella Martina Mugnano Vincenza Cerbone Maddalena Raia Giulia Scalia Ivana Kurelac Diego Luis Medina Lisa Miccio Mario Capasso Achille Iolascon Pasquale Memmolo Pietro Ferraro 《Opto-Electronic Science》 CAS 2026年第2期13-28,共16页
Virtual staining is the current state-of-the-art computational technique to cleverly enhance intracellular specificity in unstained biological samples by using convolutional neural networks(CNNs)trained on co-register... Virtual staining is the current state-of-the-art computational technique to cleverly enhance intracellular specificity in unstained biological samples by using convolutional neural networks(CNNs)trained on co-registered pairs of unstained/stained images.While effective,this approach suffers from unpredictable biases inherent to fluorescence microscopy and encounters challenges when applied to flow cytometry data as it would require accurate co-registration on a huge number of images.Here,we present a novel method that exploits for the first time a Holotomography-driven learning to completely eliminate the need for co-registration.We demonstrate that training a CNN on a stain-free dataset of 3D refractive index tomograms of flowing cells unlocks stain-free intracellular specificity for the first time in quantitative phase imaging flow cytometry.This self-supervised solution,by circumventing the critical obstacle of fluorescence co-registration,opens unprecedented perspectives for label-free,high-throughput imaging flow cytometry,offering a powerful new paradigm for advanced 2D and 3D single-cell analysis. 展开更多
关键词 quantitative phase imaging holographic tomography flow cytometry label-free computational microscopy in-silico staining deep learning
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Demands and technical developments of clinical flow cytometry with emphasis in quantitative,spectral,and imaging capabilities 认领 引用 被引量：1
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作者 Ting Zhang Mengge Gao +6 位作者 Xiao Chen Chiyuan Gao Shilun Feng Deyong Chen Junbo Wang Xiaosu Zhao Jian Chen 《Nanotechnology and Precision Engineering》 EI CAS CSCD 2022年第4期46-55,共10页
As the gold-standard method for single-cell analysis,flow cytometry enables high-throughput and multiple-parameter characterization of individual biological cells.This review highlights the demands for clinical flow c... As the gold-standard method for single-cell analysis,flow cytometry enables high-throughput and multiple-parameter characterization of individual biological cells.This review highlights the demands for clinical flow cytometry in laboratory hematology(e.g.,diagnoses of minimal residual disease and various types of leukemia),summarizes state-of-the-art clinical flow cytometers(e.g.,FACSLyricTMby Becton Dickinson,DxFLEX by Beckman Coulter),then considers innovative technical improvements in flow cytometry(including quantitative,spectral,and imaging approaches)to address the limitations of clinical flow cytometry in hematology diagnosis.Finally,driven by these clinical demands,future developments in clinical flow cytometry are suggested. 展开更多
关键词 Clinical demand Clinical flow cytometry Quantitative flow cytometry Spectral flow cytometry Imaging flow cytometry
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Correlating NAD(P)H lifetime shifts to tamoxifen resistance in breast cancer cells:A metabolic screening study with time-resolved flow cytometry 认领 引用
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作者 Samantha Valentino Karla Ortega-Sandoval +1 位作者 Kevin D.Houston Jessica P.Houston 《Journal of Innovative Optical Health Sciences》 SCIE EI CSCD 2025年第1期101-115,共15页
Time-resolved flow cytometry(TRFC)was used to measure metabolic differences in estrogen receptor-positive breast cancer cells.This specialty cytometry technique measures fluorescence lifetimes as a single-cell paramet... Time-resolved flow cytometry(TRFC)was used to measure metabolic differences in estrogen receptor-positive breast cancer cells.This specialty cytometry technique measures fluorescence lifetimes as a single-cell parameter thereby providing a unique approach for high-throughput cell counting and screening.Differences in fluorescence lifetime were detected and this was associated with sensitivity to the commonly prescribed therapeutic tamoxifen.Differences in fluorescence lifetime are attributed to the binding states of the autofluorescent metabolite NAD(P)H.The function of NAD(P)H is well described and in general involves cycling from a reduced to oxidized state to facilitate electron transport for the conversion of pyruvate to lactate.NAD(P)H fluorescence lifetimes depend on the bound or unbound state of the metabolite,which also relates to metabolic transitions between oxidative phosphorylation and glycolysis.To determine if fundamental metabolic profiles differ for cells that are sensitive to tamoxifen compared to those that are resistant,large populations of MCF-7 breast cancer cells were screened and fluorescence lifetimes were quantified.Additionally,metabolic differences associated with tamoxifen sensitivity were measured with a Seahorse HS mini metabolic analyzer(Agilent Technologies Inc.Santa Clara,CA)and confocal imaging.Results show that tamoxifen-resistant breast cancer cells have increased utilization of glycolysis for energy production compared to tamoxifen-sensitive breast cancer cells.This work is impacting because it establishes an early step toward developing a reliable screening technology in which large cell censuses can be differentiated for drug sensitivity in a label-free fashion. 展开更多
关键词 Time-resolved flow cytometry autofluorescence fluorescence lifetime breast cancer metabolism
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Effectiveness evaluation of fluorescent compensation in multicolor flow cytometry:A quantitative study 认领 引用
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作者 Long Fan Chiyuan Gao +2 位作者 Junbo Wang Xiaoye Huo Jian Chen 《Nanotechnology and Precision Engineering》 EI CAS CSCD 2025年第4期88-97,共10页
In fluorescence flow cytometry,spectral overlap among multiple fluorescent labels cannot be avoided,and thus detected fluorescent intensities need to be compensated.Although fluorescent compensation in flow cytometry ... In fluorescence flow cytometry,spectral overlap among multiple fluorescent labels cannot be avoided,and thus detected fluorescent intensities need to be compensated.Although fluorescent compensation in flow cytometry has been widely used for many years,it still lacks quantitative evaluations to validate its effectiveness.Using a home-developed nine-color fluorescence flow cytometer,this study first obtains calibration curves by assaying gradient concentrations of nine different fluorochromes individually,with the fluorescent intensities of the highest concentrations of each fluorochrome being used to obtain a spillover matrix.Mixed fluorescent solutions are analyzed by flow cytometry in which the obtained fluorescent intensities are compensated by the spillover matrix,translated to specific concentrations based on calibration curve and compared with nominal values.Three mixed solutions of Brilliant Violet 650 and Brilliant Violet 711,of Alexa Fluor 488 and PE,and of Pacific Orange,Alexa Fluor 488,and PE are tested,with fluorescent compensation being observed to reduce excessive signals due to spectral overlap.Specifically,concentration deviations(before vs after compensation)in comparison with nominal values for Brilliant Violet 711 and Alexa Fluor 488 are quantified as 40.6%vs 14.9%and 6.7%vs 1.9%,respectively.The results presented here provide a quantitative reference for fluorescent compensation that can be used to effectively address the issue of spectral overlap in fluorescence flow cytometry. 展开更多
关键词 Multicolor flow cytometry Fluorescent compensation Quantitative validation Concentration determination
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Impedance flow cytometry empowered by ConvNet algorithm to differentiate bladder cancer cells based on electro-mechanical characteristics 认领 引用
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作者 Shuaihua Zhang Zhiwen Zheng +2 位作者 Yongqi Chen Zhihong Zhang Ziyu Han 《Nanotechnology and Precision Engineering》 EI CAS CSCD 2025年第3期88-97,共10页
Bladder cancer(BC)is a common malignancy and among the leading causes of cancer death worldwide.Analysis of BC cells is of great significance for clinical diagnosis and disease treatment.Current approaches rely mainly... Bladder cancer(BC)is a common malignancy and among the leading causes of cancer death worldwide.Analysis of BC cells is of great significance for clinical diagnosis and disease treatment.Current approaches rely mainly on imaging-based technology,which requires complex staining and sophisticated instrumentation.In this work,we develop a label-free method based on artificial intelligence(AI)-assisted impedance-based flow cytometry(IFC)to differentiate between various BC cells and epithelial cells at single-cell resolution.By applying multiple-frequency excitations,the electrical characteristics of cells,including membrane and nuclear opacities,are extracted,allowing distinction to be made between epithelial cells,low-grade,and high-grade BC cells.Through the use of a constriction channel,the electro-mechanical properties associated with active deformation behavior of cells are investigated,and it is demonstrated that BC cells have a greater capability of shape recovery,an observation that further increases differentiation accuracy.With the assistance of a convolutional neural network-based AI algorithm,IFC is able to effectively differentiate various BC and epithelial cells with accuracies of over 95%.In addition,different grades of BC cells are successfully differentiated in both spiked mixed samples and bladder tumor tissues. 展开更多
关键词 Impedance flow cytometry ConvNet model Differentiation between cells Bladder cancer analysis
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Sperm chromatin condensation as an in vivo fertility biomarker in bulls:a flow cytometry approach 认领 引用 被引量：3
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作者 Marc Llavanera Jordi Ribas-Maynou +7 位作者 Ariadna Delgado-Bermúdez Sandra Recuero Rodrigo Muiño Carlos OHidalgo Carolina Tamargo Sergi Bonet Yentel Mateo-Otero Marc Yeste 《Journal of Animal Science and Biotechnology》 SCIE CAS CSCD 2022年第2期418-429,共12页
Background:Genetic selection in cattle has been directed to increase milk production.This,coupled to the fact that the vast majority of bovine artificial inseminations(AI)are performed using cryopreserved sperm,have l... Background:Genetic selection in cattle has been directed to increase milk production.This,coupled to the fact that the vast majority of bovine artificial inseminations(AI)are performed using cryopreserved sperm,have led to a reduction of fertility rates over the years.Thus,seeking sensitive and specific sperm biomarkers able to predict fertility rates is of vital importance to improve cattle reproductive efficiency.In humans,sperm chromatin condensation evaluated through chromomycin A3(CMA3)has recently been purported to be a powerful biomarker for sperm functional status and male infertility.The objectives of the present study were:a)to set up a flow cytometry method for simultaneously evaluating chromatin condensation and sperm viability,and b)to test whether this parameter could be used as a predictor of in vivo fertility in bulls.The study included pools of three independent cryopreserved ejaculates per bull from 25 Holstein males.Reproductive outcomes of each sire were determined by non-return rates,which were used to classify bulls into two groups(highly fertile and subfertile).Results:Chromatin condensation status of bovine sperm was evaluated through the combination of CMA3 and Yo-Pro-1 staining and flow cytometry.Sperm quality parameters(morphology,viability,total and progressive motility)were also assessed.Pearson correlation coefficients and ROC curves were calculated to assess their capacity to predict in vivo fertility.Sperm morphology,viability and total motility presented an area under the ROC curve(AUC)of 0.54,0.64 and 0.68,respectively(P>0.05),and thus were not able to discriminate between fertile and subfertile individuals.Alternatively,while the percentage of progressively motile sperm showed a significant predictive value,with an AUC of 0.73(P=0.05),CMA3/Yo-Pro-1 staining even depicted superior results for the prediction of in vivo fertility in bulls.Specifically,the percentage of viable sperm with poor chromatin condensation showed better accuracy and precision to predict in vivo fertility,with an AUC of 0.78(P=0.02).Conclusions:Chromatin condensation evaluated through CMA3/Yo-Pro-1 and flow cytometry is defined here as a more powerful tool than conventional sperm parameters to predict bull in vivo fertility,with a potential ability to maximising the efficiency of dairy breeding industry. 展开更多
关键词 Bull Chromatin Chromomycin A3 Condensation Fertility Flow cytometry Sperm
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Flow cytometry for the assessment of animal sperm ntegrity and functionality: state of the art 认领 引用 被引量：3
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作者 Md. Sharoare Hossain Anders Johannisson +3 位作者 Margareta Wallgren Szabolcs Nagy Amanda Pimenta Siqueira Heriberto Rodriguez-Martinez 《Asian Journal of Andrology》 SCIE CAS CSCD 2011年第3期406-419,511,共14页
Flow cytometry is now a recognized methodology within animal spermatology, and has moved from being a research tool to become routine in the assessment of animal semen destined to breeding. The availability of ＇bench... Flow cytometry is now a recognized methodology within animal spermatology, and has moved from being a research tool to become routine in the assessment of animal semen destined to breeding. The availability of ＇bench-top＇ flow cytometers and of newer and versatile markers for cell structure and function had allowed the instrumentation to measure more sperm parameters, from viability to reactiveness when exposed to exogenous stimuli, and to increase our capabilities to sort spermatozoa for potential fertilizing capacity, or chromosomal sex. The present review summarizes the state of the art regarding flow cytometry applied to animal andrology, albeit keeping an open comparative intent. It critically evaluates the present and future capabilities of flow cytometry for the diagnostics of potential fertility and for the development of current reproductive technologies such as sperm freezing, sperm selection and sperm sorting. The flow cytometry methods will probably further revolutionize our understanding of the sperm physiology and their functionality, and will undoubtedly extend its application in isolating many uncharacterized features of spermatozoa. However, continuous follow-up of the methods is a necessity owing to technical developments and the complexity of mapping spermatozoa. 展开更多
关键词 animals fertility flow cytometry semen analysis spermatozoa sperm functionality sperm intactness
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Analysis of DNA Ploidy, Cell Cycle and Ki67 Antigen in Nasopharyngeal Carcinoma by Flow Cytometry 认领 引用 被引量：11
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作者 石小燕 袁响林 +2 位作者 陶德定 龚建平 胡国清 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 2005年第2期198-201,共4页
Summary: The expression of DNA ploidy, the cell cycle and Ki67 antigen in nasopharyngeal carcinoma (NPC) were studied and their relationship with the clinical biological behaviors and prognosis of NPC was evaluated. B... Summary: The expression of DNA ploidy, the cell cycle and Ki67 antigen in nasopharyngeal carcinoma (NPC) were studied and their relationship with the clinical biological behaviors and prognosis of NPC was evaluated. Biopsied specimens of NPC were made into cell suspension. By using cytometric double labeling Ki67 and DNA method, the expression of DNA ploidy, the cell cycle and Ki67 antigen were analyzed. The patients were followed-up for about 3 years and the relationship between the above-mentioned parameters and the clinical biological behavior and prognosis of NPC were evaluated. Of the 62 cases of NPC, the DNA aneuploid accounted for 29.03 %. The S phase cells accounted for 0 to 54 % in the cell cycle and the positive expression of Ki67 ranged from 0 to 52 %. There were 40 cases of LPI (64.5 %) including 15 negative cases and 22 cases of HPI (35 5 %) respectively. The DNA anueploid content was positively related to the S phase cells. The patients having a low expression of Ki67 or DNA aneuploid in tumor cells were not sensitive to chemotherapy, liable to metastasis to distant organs and had a poor prognosis, while Ki67 showed no correlation with DNA ploidy and the cell cycle. It was suggested that DNA ploidy and Ki67 could be used as an independent and objective marker to evaluate the radiosensitivity and prognosis of NPC. 展开更多
关键词 nasopharyngeal carcinoma DNA ploidy cell cycle Ki67 antigen flow cytometry
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Magnetic cell sorting and flow cytometry sorting methods for the isolation and function analysis of mouse CD^4+CD25^+Treg cells 认领 引用 被引量：4
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作者 Hang YAN Chen-guang DING +6 位作者 Pu-xun TIAN Guan-qun GE Zhan-kui JIN Li-ning JIA Xiao-ming DING Xiao-ming PAN Wu-jun XUE 《Journal of Zhejiang University-SCIENCE B》 SCIE CAS 2009年第12期928-932,共5页
Objective:In this paper we compared the two methods of cell sorting(magnetic cell sorting and flow cytometry sorting)for the isolation and function analysis of mouse CD4+CD25+regulatory T(Treg)cells,in order to inform... Objective:In this paper we compared the two methods of cell sorting(magnetic cell sorting and flow cytometry sorting)for the isolation and function analysis of mouse CD4+CD25+regulatory T(Treg)cells,in order to inform further studies in Treg cell function.Methods:We separately used magnetic cell sorting and flow cytometry sorting to identify CD4+CD25+Treg cells.After magnetic cell separation,we further used flow cytometry to analyze the purity of CD4+CD25+Treg cells,trypan blue staining to detect cell viability,and propidium iodide(PI)staining to assess the cell viability.We detected the immune inhibition of CD4+CD25+Treg cells in the in vitro proliferation experiments.Results:The results showed that compared to flow cytometry sorting,magnetic cell sorting took more time and effort,but fewer live cells were obtained than with flow cytometry sorting.The CD4+CD25+Treg cells,however,obtained with both methods have similar immunosuppressive capacities.Conclusion:The result suggests that both methods can be used in isolating CD4+CD25+Treg cells,and one can select the best method according to specific needs and availability of the methodologies. 展开更多
关键词 CD4+CD25+Treg cells Flow cytometry sorting Magnetic cell sorting
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Recent advances in fuorescence-based in vivo flow cytometry 认领 引用 被引量：3
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作者 Kai Pang Bobo Gu +3 位作者 Feng Liu Mingli Dong Lianqing Zhu Xunbin Wei 《Journal of Innovative Optical Health Sciences》 SCIE EI 2019年第6期1-10,共10页
The fuorescence-based in vivo flow cytometry(IVFC)is an emerging tool to monitor eirculating cells in vivo.As a noninvasive and real-timne diagnostic technology,the fluorescence based IVFC allows long-term monitoring ... The fuorescence-based in vivo flow cytometry(IVFC)is an emerging tool to monitor eirculating cells in vivo.As a noninvasive and real-timne diagnostic technology,the fluorescence based IVFC allows long-term monitoring of circulating cells without changing their native biological environment.It has been applied for various biological applications(eg,monitoring circulating tumor cells).In this work,we will review our recent works on fluorescence-based IVFC.The operation principle and typical biological applications will be introduced.In addition,the recent advances in IVFC flow cytometry based on photoacoustic effects and other label free detection methods such as imaging based methods,difuse-light methods,hybrid multimodality methods and multispectral methods are also summarized. 展开更多
关键词 In vrivo flow cytometry cireulating tumor cells(CTCs) CTC clusters nanoparticles fluorescence
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Characterization of ploidy levels in Chrysanthemum L. by flow cytometry 认领 引用 被引量：3
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作者 Yue-ping Ma Jiang-xue Wei +2 位作者 Zhi-yang Yu Bing Qin Si-lan Dai 《Journal of Forestry Research》 SCIE CAS CSCD 2015年第3期771-775,共5页
Analyzing the ploidy levels of plants is important for identifying species, selecting parental lines, identifying the relationships between species, and determining evolutionary patterns. The genus Chrysanthemum is wi... Analyzing the ploidy levels of plants is important for identifying species, selecting parental lines, identifying the relationships between species, and determining evolutionary patterns. The genus Chrysanthemum is widely distributed throughout the world and exhibits different ploidy levels. We used flow cytometry to analyze the ploidy levels of nine species of Chrysanthemum L. collected from different regions and geographical locations in China. Three diploids from Henan and Wuhan provinces corresponded to Chrysanthe- mum lavandulifolium and two species of C. nankingense, while three tetraploids from various regions corresponded to C. indicum and two species of C. chanetii. Two hexaploids corresponding to C. vestitum were collected at Funiu moun- tain （Henan province）, and C. zawadskii was collected at Huangshan mountain （Anhui province）. We found that OTTO extraction buffer was suitable for extracting nuclei from most species, apart from C. zawadskii. Flow cytometry proved to bea simple, rapid, and highly accurate method for identifying ploidy levels in Chrysanthemum species. 展开更多
关键词 Chrysanthemum L. Flow cytometry - OTTOextraction buffer Ploidy identification
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Specific Detection of Toxigenic Vibrio cholerae Based on in situ PCR in Combination With Flow Cytometry 认领 引用 被引量：2
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作者 LI ZHU JUN-PENG CAI +1 位作者 QING CHEN SHOU-YI YU 《Biomedical and Environmental Sciences》 SCIE CAS 2007年第1期64-69,共6页
Objective To develop an in situ PCR in combination with flow cytometry （ISPCR-FCM） for monitoring cholera toxin positive Vibrio cholerae. Methods In running this method, 4% paraformaldehyde was used to fix the Vibri... Objective To develop an in situ PCR in combination with flow cytometry （ISPCR-FCM） for monitoring cholera toxin positive Vibrio cholerae. Methods In running this method, 4% paraformaldehyde was used to fix the Vibrio cholerae cells and 1 mg/mL lysozyme for 20 min to permeabilize the cells. Before the PCR thermal cycling, 2.5% glycerol was added into the PCR reaction mixture in order to protect the integrality of the cells. Results A length of 1037bp DNA sequence was amplified, which is specific for the cholera toxin gene （ctxAB gene）. Cells subjected to ISPCR showed the presences of ctxAB gene both in epifluorescence microscopy and in flow cytometric analysis. The specificity and sensitivity of the method were investigated. The sensitivity was relatively low （10^5 cells/mL）, while the specificity was high. Conclusion We have successfully developed a new technique for detection of toxigenic Vibrio cholerae strains. Further study is needed to enhance its sensitivities. ISPCR-FCM shows a great promise in monitoring specific bacteria and their physiological states in environmental samples. 展开更多
关键词 Vibrio cholerae Detection technique in situ PCR Flow cytometry
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Advances in small animal mesentery models for in vivo flow cytometry,dynamic microscopy,and drug screening 认领 引用 被引量：2
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作者 Ekaterina I Galanzha Valery V Tuchin Vladimir P Zharov 《World Journal of Gastroenterology》 SCIE CAS 2007年第2期192-218,共27页
Using animal mesentery with intravital optical microscopy is a well-established experimental model for studying blood and lymph microcirculation in vivo. Recent advances in cell biology and optical techniques provide ... Using animal mesentery with intravital optical microscopy is a well-established experimental model for studying blood and lymph microcirculation in vivo. Recent advances in cell biology and optical techniques provide the basis for extending this model for new applications, which should generate significantly improved experimental data. This review summarizes the achievements in this specific area, including in vivo label-free blood and lymph photothermal flow cytometry, super-sensitive fluorescence image cytometry, light scattering and speckle flow cytometry, microvessel dynamic microscopy, infrared (IR) angiography, and high-speed imaging of individual cells in fast flow. The capabilities of these techniques, using the rat mesentery model, were demonstrated in various studies; e.g., real-time quantitative detection of circulating and migrating individual blood and cancer cells, studies on vascular dynamics with a focus on lymphatics under normal conditions and under different interventions (e.g. lasers, drugs, nicotine), assessment of lymphatic disturbances from experimental lymphedema, monitoring cell traffic between blood and lymph systems, and high-speed imaging of cell transient deformability in flow. In particular, the obtained results demonstrated that individual cell transportation in living organisms depends on cell type (e.g., normal blood or leukemic cells), the cell’s functional state (e.g., live, apoptotic, or necrotic), and the functional status of the organism. Possible future applications, including in vivo early diagnosis and prevention of disease, monitoring immune response and apoptosis, chemo- and radio-sensitivity tests, and drug screening, are also discussed. 展开更多
关键词 Lymph microcirculation Transmission digital microscopy Rat mesentery Flow cytometry Photothermal technique
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DETECTION OF PLATELET-DERIVED MICROPARTICLES USING FLOW CYTOMETRY AND ITS CLINICAL APPLICATION 认领 引用 被引量：2
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作者 崔巍 马文新 +1 位作者 林其燧 韩晔华 《Chinese Medical Sciences Journal》 CAS 2003年第1期26-30,共5页
Objective.To establish a flow cytometric internal standard method for counting platelet-derived microparti-cles(PMPs)and to study its clinical significance. Methods. PMPs suspension(platelet poor plasma,PPP) was extra... Objective.To establish a flow cytometric internal standard method for counting platelet-derived microparti-cles(PMPs)and to study its clinical significance. Methods. PMPs suspension(platelet poor plasma,PPP) was extracted by gradual centrifugation. According to the size of PMPs,3 μm and 0.8μm latex beads were used as internal standards for the quantitation. PMPs were counted by adjusting flow cytometric discrimination and voltage of forward scatter and side scatter. Results. In 30 healthy donors,the average concentration of resting PMPs was(1.2×105±5.7×104 )/ml and that of activated PMPs was(1.6×106±9.1×105)/ml. Compared with healthy donors,PMPs mean value was significantly higher(P0.05)] .Conclusions. PMPs is a useful indicator in monitoring platelet activation,and plays an important role in thrombotic disease. By flow cytometric internal standard method,PMPs can be counted rapidly and accurately,which may be very helpful in interlaboratory comparative studies. 展开更多
关键词 platelet microparticles platelet activation flow cytometry
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A Sensitive and High-Throughput Flow Cytometry-Based Assay for Measuring Antibody Neutralization of Human Adenovirus Type 3 认领 引用 被引量：1
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作者 Zhenwei Liu Xingui Tian +3 位作者 Wenkuan Liu Yuting Xian Weilue Chen Rong Zhou 《Virologica Sinica》 SCIE CAS CSCD 2021年第3期537-544,共8页
The assessment of neutralization activity is an important step in the evaluation of neutralizing antibodies (NAbs).The traditional methods for measuring the antibody neutralization of human adenovirus type 3 (HAd V-3)... The assessment of neutralization activity is an important step in the evaluation of neutralizing antibodies (NAbs).The traditional methods for measuring the antibody neutralization of human adenovirus type 3 (HAd V-3) are the microneutralization (MN) assay,which has insufficient sensitivity,and the plaque reduction neutralization test (PRNT),which is not suitable for high-throughput screening.Herein,we describe the development of a flow cytometry-based neutralization(FCN) assay for measuring the neutralization of sera,cell culture supernatants,and chimeric antibodies against HAd V-3 on the basis of a recombinant HAd V-3 (r HAd V-3) construct expressing the enhanced green fluorescent protein (EGFP).For flow cytometry-based assays,the optimal cell confluence was determined as 90%,and the virus was titrated using the assay.The established FCN assay follows the percentage law and an optimal MOI of not less than 5 9 10~(-4)was determined by using a purified chimeric antibody.In addition,comparison of the anti-HAd V-3 NAb titers of 72 human serum samples by the MN and FCN assays,showed that both assays correlated strongly with each other.Our FCN assay was an improvement over the MN assay because the observation period was reduced from 3 to 1 days and data analysis could be performed objectively and robotically.Importantly,the newly established FCN assay allows measurement of the neutralization activity of chimeric antibodies expressed in cell culture supernatants.Thus,this sensitive and highthroughput FCN assay is a useful alternative to the MN assay for measuring the antibody neutralization of HAd V-3 and for screening anti-HAd V-3 NAbs in cell culture supernatants. 展开更多
关键词 Sensitive High-throughput Flow cytometry Neutralization Adenovirus
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Application of CD45/SSC Gating Multiparameter Flow Cytometry in the Classification of Acute Leukemia——An Analysis of 139 Cases 认领 引用 被引量：2
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作者 黎纬明 陈智超 +1 位作者 刘仲萍 邹萍 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 2001年第3期209-211,共3页
In order to study the significance of flow cytometry immunophenotyping in the diagnosis of acute leukemia, CD45/SSC gating multiparameter flow cytometry (FCM) was utilized to analyze the immunophenotypes of 139 cases ... In order to study the significance of flow cytometry immunophenotyping in the diagnosis of acute leukemia, CD45/SSC gating multiparameter flow cytometry (FCM) was utilized to analyze the immunophenotypes of 139 cases of acute leukemia. 139 cases of acute leukemia were enrolled in our hospital from April 1998 to April 2000. Morphological analysis and FCM immunophenotypic tests were conducted on all cases. Our results showed that CD45/SSC gating multiparameter flow cytometry immunophenotyping could reflect the origin of leukemic cells specifically. It is one of the important methods for the diagnosis of ALL, AML, and HAL. CD45/SSC gating multiparameter FCM analysis is a good technique for immunophenotyping. FCM immunophenotypic analysis can help improve the diagnosis and classification of acute leukemia, and extend the use of FCM in clinical practice. 展开更多
关键词 acute leukemia immunophenotype flow cytometry CD45
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CLINICAL SIGNIFICANCE OF THE DETECTION OF MINIMAL RESIDUAL DISEASE IN CHILDHOOD B-ALL BY FLOW CYTOMETRY 认领 引用 被引量：1
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作者 沈立松 赵惠君 +3 位作者 徐翀 吴政宏 汤静燕 陈静 《Journal of Shanghai Second Medical University(Foreign Language Edition)》 2003年第2期100-104,111,共5页
Objective To detect the minimal residual disease in children with B-ALL and to evaluate its clinical significance by flow cytometry. Methods 58 childhood B-ALL cases were enrolled into this study and 33 MRD analyses w... Objective To detect the minimal residual disease in children with B-ALL and to evaluate its clinical significance by flow cytometry. Methods 58 childhood B-ALL cases were enrolled into this study and 33 MRD analyses were performed after remission induction therapy.Four-color combinations of fluorochrome labeled monoclonal antibodies against lymphocyte lineage related phenotypes were used to analyze leukemic cells with flow cytometry.The cells from normal bone marrow were used as controls.The combinations of phenotypes that reflect the antigen expression differences between leukemic and normal bone marrow cells on flow cytometry were considered to be the effective phenotype combinations in the first step screening.The effective phenotype combinations were then used to monitor MRD during the disease course after therapy began. Results 58 cases of childhood B-ALL were screened for MRD effective phenotype combinations.The effective phenotype combinations were identified in 89.7% of B-ALL cases in this study.Four-color phenotype combinations were composed of CD10/CD34/CD19 plus another effective marker such as CD38,CD58,CD66c,CD21.The senstitivity of this method was 0.01%,much higher than that of microscopic inspection.In 8 cases,their bone marrow microscopic inspection results showed no remaining leukemic cells;but with flow cytometry,the percentage of leukemic cells were 5.66%,0.36%,1.43%,0.069%,1.55%,2.7%,0.028% and 0.015%,respectively.In risk stratification,all these MRD positive cases were classified into high risk group for relapse and 1 case showed early relapse within 6 months. Conclusion The application of flow cytometry in MRD measurement can significantly improve the sensitivity of detection of remained leukemic cells in childhood B-ALL,and can provide more accurate information on disease progression as well as the efficacy of therapy,thus facilitate future treatment decisions and follow ups. 展开更多
关键词 B-ALL immunophenotyping minimal residual disease flow cytometry
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Identification of Prorocentrum minimum and Takayama pulchella by fluorescence in situ hybridization through epifluorescence microscopy and flow cytometry 认领 引用
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作者 HOU Jianjun LAI Hongyan +1 位作者 HUANG Bangqin CHEN Jixin 《Acta Oceanologica Sinica》 SCIE CAS 2009年第2期103-114,共12页
Partial rDNA sequences of Prorocentrum minimum and Takayama pulchella were amplified, cloned and sequenced, and these sequence data were deposited in the GenBank. Eight oligonucleotide probes （DNA probes） were desig... Partial rDNA sequences of Prorocentrum minimum and Takayama pulchella were amplified, cloned and sequenced, and these sequence data were deposited in the GenBank. Eight oligonucleotide probes （DNA probes） were designed based on the sequence analysis. The probes were employed to detect and identify P. minimum and T. pulchella in unialgal and mixed algal samples with a fluorescence in situ hybridization method using flow cytometry. Epifluorescence micrographs showed that these specific probes labeled with fluorescein isothiocyanate entered the algal cells and bound to target sequences, and the fluorescence signal resulting from whole-cell hybridization varied from probe to probe. These DNA probes and the hybridization protocol we developed were specific and effective for P. minimum and T. pulchella, without any specific binding to other algal species. The hybridization efficiency of different probes specific to P. minimum was in the order： PM18S02 PM28S02 〉 PM28S01 〉PM18S01, and that of the probes specific to T. pulcheUa was TP18S02 TP28S01 〉 TP28S02 〉TP18S01. The different hybridization efficiency of the DNA probes could also be shown in the fluorescent signals between the labeled and unlabeled cells demonstrated using flow cytometry. The DNA probes PM18S02, PM28S02, TP18S02 and TP28S01, and the protocol, were also useful for the detection of Mgae in natural samples. 展开更多
关键词 oligonucleotide DNA probes Prorocentrum minimum Takayama pulcheUa fluorescence in situ hybridization flow cytometry
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Selection of anticoagulant solution to the hemolymph of Chlamys farreri by transmission electron microscropy and flow cytometry 认领 引用
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作者 陈慕雁 Yang Hongsheng Shi Fangfang 《High Technology Letters》 EI CAS 2007年第2期221-224,共4页
Mixtures of hemolymph from Chlamys farreri with three different anticoagulant solutions were incu-bated for an hour in vitro, then the ultrastructural alterations of hemocytes were observed, and the aggre-gation rate ... Mixtures of hemolymph from Chlamys farreri with three different anticoagulant solutions were incu-bated for an hour in vitro, then the ultrastructural alterations of hemocytes were observed, and the aggre-gation rate was analyzed by using transmission electron microscropy and flow cytometry respectively. The results showed that Formula 3 （glucose 20.8 g L^-1; EDTA 20mM; sodium chloride 20 g L^-1; Tris-HCI 0.05M;pH 7.4）was the desirable anticoagulant solution for C. farreri hemocytes. Further phagocytosis assay showed that no obvious negative effect was given to the hemocyte phagocytic activity when using Formula 3 as the anticoagulant solution. 展开更多
关键词 hemolymph anticoagulant solution TEM flow cytometry Chlamys farreri
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