• Title, Summary, Keyword: flow cytometry

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Viability Assessment of Fresh and Frozen-thawed Dog Spermatozoa by Flow Cytometry (Flow Cytometry에 의한 개 신선정액과 동결정액의 생존성 분석)

  • Hong Y. M.;Kim Y. J.;Yu I.;Ji D. B.;Kim M. S.
    • Reproductive and Developmental Biology
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    • v.28 no.3
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    • pp.167-172
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    • 2004
  • This study was performed to examine the correlations among dog sperm viabilities evaluated by flow cytometry, by microscopic evaluation (ME), by carbo-xifluorescein diacetate and propidium iodide (CFDA/PI) and by hypoosmotic swelling (HOS) test. Semen were collected from 5 dogs ranging in age from 2 to 4 years. Each ejaculate was divided into 3 aliquots and different proportions of freeze-killed cells were added to each aliquot (1:0, 1:1 and 1:3). In the other experiment, semen was extended with Sweden extender containing 5% glycerol and equex STM paste, and frozen using liquid nitrogen vapor. Fresh and frozen-thawed dog sperm viability were assessed by flow cytometry using PI staining method. The accuracy of flow cytometry was evaluated by comparing with other classic assessments, microscopic evaluation, epifluorescence microscopic analysis using CFDA/PI, and HOS test. High correlations of sperm viabilities were found among flow cytometry, epifluorescence evaluation, HOS test (p<0.01) in fresh semen. Especially, sperm viability assessed by HOS test was highly correlated with viability by flow cytometry in all the ratios of live and dead spermatozoa, 1:0, 1:1 and 1:3 (p<0.01). The viability evaluated by ME were significantly correlated with that by flow cytometry in ratios of 1:0 and 1:3 (p<0.05) however, there was no significance in ratio of 1:1. The viability evaluated by C/p were highly correlated with that by flow cytometry in ratio of 1:0 and 1:1 (p<0.01) and significantly correlated in ratio of 1:3 (p<0.05). In frozen-thawed spermatozoa, the viability determined by HOS test was considerably correlated with that by flow cytometry (p<0.01). There was significant correlation between the viabilities by ME and by flow cytometry (p<0.05). But the viability evaluated by CFDA/PI was not correlated with viability by flow cytometry. The result from this study validate the use of flow cytometry as a precise method for assessing the viability of fresh and frozen-thawed dog spermatozoa.

Microfluidic Flow Cytometry: Principles of Cell Analysis and Applications

  • Shin, Se-Hyun
    • International Journal of Vascular Biomedical Engineering
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    • v.4 no.2
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    • pp.1-6
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    • 2006
  • Microsystems create new opportunities for conventional cell analysis by combining microfluidics and flow cytometry. This article describes recent developments in conventional flow cytometers and related microfluidic flow cytometers to detect, analyze, and sort cells or particles. Flow cytometry strongly consisted of fluidics, optics and electronics requires a large space to equip various components, which are mostly the fluidic components such as compressor, fluid handling system. Adopting microfluidics into flow cytometry enables volume- and power-efficient, inexpensive and flexible analysis of particulate samples. In this paper, we review various efforts that take advantage of novel techniques to build microfluidic cell analysis systems with high-speed analytical capability. Highly integrated microfluidic cytometry shows great promise for basic biomedical and pharmaceutical research, and robust and portable point-of-care devices could be used in clinical settings.

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Application of Flow Cytometry to Monitoring of Liposomal Restructuring Induced by Listeria monocytogenes

  • Kim, Hyung-Joo;Bennetto, H.-Peter;Halablab, Mahmoud-A.
    • Journal of Microbiology and Biotechnology
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    • v.14 no.5
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    • pp.1099-1102
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    • 2004
  • Liposomal restructuring induced by hemolytic Listeria monocytogenes was investigated by using flow cytometry. When added to calcein-entrapped liposomes, hemolytic, but not non-hemolytic, Listeria monocytogenes were able to induce reformation of vesicles. Such restructuring of liposomes was easily monitored by flow cytometry. Electron microscopy also indicated major changes in the challenged liposomal structures. The preliminary results described may offer a simple and fast method for monitoring liposomal restructuring and for differentiating between hemolytic and non-hemolytic bacteria.

Histopathological Observation and Flow Cytometry Analysis of Testicular Atrophy Induced by 2-Bromopropane On the Sprague-Dawley Rat (2-Bromopropane에 의한 유발된 Sprague-Dawley 랫트의 고환위축의 병리학적 관찰 및 Flow Cytometry를 이용한 검사)

  • 손화영;강부현;조성환;차신우;노정구
    • Toxicological Research
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    • v.14 no.2
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    • pp.143-149
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    • 1998
  • This study was carried out to evaluate the testicular toxicity of 2-bromopropane (2-BP), which recently caused occupational intoxication on the reproductive and hematopoietic system in Koreans, using light microscopy, immunohistochemistry and flow cytometry. 10 weeks old male Sprague-Dawley (SD) rats were treated with 0.5 g/㎏/day of 2-BP orally for 8 consecutive weeks. The testes of the rats were vascularly perfused with Karnovsky's solution or immersed in Bouin's solution, embedded in plastic and evaluated with light microscopy. And relative proportions of haploid, diploid, and tetra-ploid states of DNA ploidy in the testicular cell suspensions of the SD rats were examined by flow cytometry. 2-BP induced severe testicular atrophy, depletion and degeneration of spermatogonia, spermatocytes, and spermatids and mild hyperplasia of Leydig cells without significant morphological changes. The Leydig cell hyperplasia was confirmed by immunohistochemistry using proliferating cell nuclear antigen (PCNA). The immunopositive cells against PCNA were observed in the nuclei oj some interstitial cells. Relative proportions of haploid states of DNA ploidy decreased in the atrophic testicular cell suspensions comparing with those of the control. In conclusion, 2-BP induced testicular atrophy with Leydig cell hyperplasia as examined by histopathology, immunohistochemistry and DNA flow cytometry.

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Multiparameter Flow Cytometry: Advances in High Resolution Analysis

  • O'Donnell, Erika A.;Ernst, David N.;Hingorani, Ravi
    • IMMUNE NETWORK
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    • v.13 no.2
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    • pp.43-54
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    • 2013
  • Over the past 40 years, flow cytometry has emerged as a leading, application-rich technology that supports high-resolution characterization of individual cells which function in complex cellular networks such as the immune system. This brief overview highlights advances in multiparameter flow cytometric technologies and reagent applications for characterization and functional analysis of cells modulating the immune network. These advances significantly support highthroughput and high-content analyses and enable an integrated understanding of the cellular and molecular interactions that underlie complex biological systems.

Detection of Escherichia coli Using Flow Cytometry (유세포 분석기를 이용한 대장균 검출에 관한 연구)

  • Kim, Ji-Hye;Park, Sang-Won;Cho, Young Sik
    • Journal of Environmental Science International
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    • v.26 no.1
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    • pp.11-21
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    • 2017
  • In this study, bacterial growth was assessed by flow cytometry analysis of fluorescent probes-stained bacteria. Flow cytometry has many advantages of rapid analytical time, a low standard deviation, and highly sensitive detection of live and Dead E.coli over colony forming assay. When untreated bacteria were stained by using Thiazole Orange (TO) and Propidium Iodide (PI), double staining had a short analytical time as compared with that of single staining while its error rate was similar to that of single staining. Through double staining experiments, it was determined that optimal concentrations for TO and PI staining were 420 nM and $9.6{\mu}M$, respectively.

Generation of sheath-free particle beam: application to micro-flow cytometry (외피유체 없이 입자 빔의 발생: 유세포 분류기 응용)

  • Kim, Young-Won;Yoo, Jung-Yul
    • 한국전산유체공학회:학술대회논문집
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    • pp.581-584
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    • 2008
  • A generation of a particle beam is the key technique in a flow cytometry that measures the fluorescence and light scattering of individual cell and other particulate or molecular analytes in biomedical research. Recent methods performing this function require a laborious and time-consuming assembly. In the present work, we propose a novel device for the generation of an axisymmetrical focusing beam of microparticles (3-D focusing) in a single capillary without sheath flows. This work uses the concept that the particles migrate toward the centerline of the channel when they lag behind the parabolic velocity profile. Particle focusing of spherical particles was successfully made with a beam diameter of about 10 ${\mu}$m. Proposed device provides crucial solutions for simple and innovative 3-D particle focusing method for the applications to the MEMS-based micro-flow cytometry. We believe that this device can be utilized in a wide variety of applications, such as biomedical/ biochemical engineering.

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Colchicine-Induced Polyploidy and It's Agronomic Characters in Bupleurum falcatum (배수체 작성에 따른 시호 작물 특성)

  • Son, Tae-Kwon;Lee, Sang-Chul;Chung, Il-Kyung
    • Korean Journal of Medicinal Crop Science
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    • v.16 no.1
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    • pp.39-43
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    • 2008
  • The effect of colchicine treatment on the agronomic performance and polyploid formation of Bupleurum falcatum using flow cytometry technique was investigated. The roots of 4-leaf stage plants were treated with colchicine (0.5%) for 3, 6, 12, and 24 hours and then transplanted in the field. Agronomic characters (survival rate, plant height, chlorophyll content, bolting rate) were recorded at 4 weeks and 6 months after transplanting while flow cytometry technique was conducted for determination of polyploid formation. Flow cytometry technique revealed polyploid nuclear DNA formation in colchicine treated plants. The highest number of polyploids was obtained at the shortest colchicine treatment time indicating an inverse relationship between colchicine treatment time and polyploid formation. Results also showed that survival and bolting rates were inversely related with the treatment time while plant height and chlorophyll were not significantly affected by the treatment. This study showed a convenient method for determination of colchicine-induced polyploid in B. falcatum and its superior agronomic performance at shorter treatment time.

Prognostic Significance of Peripheral Blood Flow Cytometry Parameters in Patients with Non-Metastatic Breast Cancer

  • Engin, Huseyin;Bilir, Cemil;Tekin, Ishak Ozel
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.12
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    • pp.7645-7649
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    • 2013
  • Background: Immune functions and their relation to prognosis in breast cancer patients have become areas of great interest in recent years. Correlations between survival outcomes and peripheral blood flow cytometry parameters are therefore of interest. Here we focused on patients with non-metastatic breast cancer (BC). Materials and Methods: A total of 29 patients with pathological confirmed breast carcinoma and flow cytometry data were assessed for overall survival (OS) and progression free survival (PFS). Results: The median age of the patients was 54 years (range, 29-83). Multivariate analysis revealed that OS was significantly associated with absolute cytotoxic T cell count (95%CI, coef 2.26, p=0.035), tumor size (95%CI, coef -14.5, p 0.004), chemotherapy (95%CI, coef 12.9, p 0.0001), MFI of CD4 (95%CI, coef -5.1, P 0.04), MFI of HLA DR (95%CI, coef -5.9, p 0.008) and tumor grade (95%CI, coef -13, P 0.049) with R-Sq(adj)=67%. Similar findings were obtained for PFS. Conclusions: OS and PFS were significantly associated with tumor grade, tumor size, chemotherapy, MFI of CD4, HLA DR and absolute cytotoxic T cell count. The study revealed that MFI of basic CD markers and absolute cytotoxic T cell number may be a prognostic factors in women with non-metastatic BC.

Comparison of Two Fluorescent Stain Methods for Jeju Black Cattle Spermatozoa Viability Assessment by Using Flow Cytometry (제주흑우 정자 생존성 평가를 위해 flow cytometry를 사용한 두가지 형광 염색법의 비교)

  • Shin, Sang-Min;Park, Seol-Hwa;Son, Jun-Gyu;Cho, In-Cheol;Seong, Pil-Nam;Kim, Nam-Young;Woo, Jai-Hoon;Shin, Moon-Cheol;Park, Nam-Geon
    • Journal of Embryo Transfer
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    • v.32 no.3
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    • pp.221-226
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    • 2017
  • Spermatozoa viability can be assessed by microscopy, flow cytometry, and other methods using fluorescent stain. Flow cytometry can be used to examine the morphological and functional characteristics of spermatozoa in a short time. The purpose of this study was to compare the viability of cryopreserved spermatozoa in Jeju black cattle by two dual fluorescent stain methods. Semen of Jeju black cattle raised in Subtropical Livestock Research Institute, National Institute of Animal Science, RDA were collected with artificial vaginal technique. Sperm was diluted with $Triladyl^{(R)}$-egg yolk diluent and then was performed cryopreservation. There was no significant difference in viability of spermatozoa according to the two dual fluorescent stain methods. However, when the distribution of spermatozoa according to the staining method was compared, the spermatozoa group stained with 6-CFDA/PI was more clearly distinguished than the spermatozoa group stained with calcein AM/PI.