• Title/Summary/Keyword: flow cytometry analysis

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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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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.

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.

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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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.

Enumeration of Weissella cibaria phage with cytometry, epifluorescence microscopy, and plaque assay (유세포분석기, 형광현미경, 용균반검사 분석을 이용한 Weissella cibaria 박테리오파지 정량분석 및 상관관계분석)

  • Park, Won Jeong;Lim, Ga-Yeon;Park, Jong-Hyun
    • Korean Journal of Food Science and Technology
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    • v.50 no.2
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    • pp.244-247
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    • 2018
  • Quantitative analysis for non-host infection bacteriophage was conducted for their enumeration. Flow cytometry and epifluorescence microscopy (EPM) were selected as counting methods. Correlation analysis was performed based on the plaque assay method on the existing host infection and consisted of Pearson correlation statistical analysis, regression analysis, and difference analysis. Analyses of 12 samples with flow cytometry and plaque assay methods showed that there was a correlation of 96.7% with Pearson correlation value r=0.967, $R^2$ 0.9352, and difference value of 1.063. Analyses of 12 samples with EPM and plaque assay methods showed that there was a correlation of 99.0% with Pearson correlation value r=0.990, $R^2$ 0.9811, and difference value of 1.605. Therefore, flow cytometry and epifluorescence microscopy would be effective for enumeration of Weissella cibaria bacteriophage with plaque assay.

Diagnostic Value of Flow Cytometric DNA Analysis in the Evaluation of Effusions (체강삼출액의 진단에 있어서 유세포분석에 의한 DNA 함량 측정의 유용성)

  • Lee, Ji-Shin;Juhang, Sang-Woo
    • The Korean Journal of Cytopathology
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    • v.8 no.1
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    • pp.20-26
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    • 1997
  • The specificity of cytologic examination in effusions is high but the sensitivity is low. Therefore, various ancillary methods for the detection of malignant cells in effusions have been proposed. The presence of an aneuploid cell population is generally considered diagnostic of malignancy. The purpose of this study is to determine whether the routine use of flow cytometry adds to standard cytologic evaluation in effusions. We did flow cytometric DNA analysis in 76 effusions(28 malignant and 48 benign fluids). All the 48 benign effusions were diploid. There were 12(42.9%) aneuploid and 16(67.1%) diploid malignant effusions. Based on these results flow cytometric DNA analysis had a sensitivity of 42.9% and a specificity of 100%. These results suggest that flow cytometric DNA analysis may be a useful adjunct to conventional cytology, but its principal limitation is us relatively low sensitivity.

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Cell Viability and Flow Cytometry Analysis of a Novel Antitumor Agent, Heptaplatin in Human Melanoma Cell Line, SK-MEL-28 (신규항암제인 Heptaplatin의 인체 흑색종세포(SK-MEL-28)에 대한 세포생존률 및 유세포 분석)

  • 최수라;명평근
    • YAKHAK HOEJI
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    • v.47 no.6
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    • pp.345-351
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    • 2003
  • Heptaplatin, cis-Malonato[(4R,5R)-4,5-bis(aminomethyl)-2-isopropyl-1,3-dioxolane]platinum(II), is a novel platinum-based antitumor agent with clinical potential against human stomach cancer and the 3rd generation of the cisplatin. This study was performed to study how cisplain, heptaplatin and sunpla which is a mixture of heptaplatin and mannitol (w: w=l : 2) affect cell viability of SK-MEL-28 human melanoma cell line. Heptaplatin ($IC_{50}$/; 95.35 $\mu$M) and sunpla ($IC_{50}$/; 10.95 11M) were less effect than cisplatin (IC $_{50}$; 10.92 $\mu$M) on the SK-MEL-28 cells. By cell cycle analysis using flow cytometry, it was identified that the cells were arrested at G2/M phase by cisplatin, heptaplatin and sunpla, and percentage of cell death group was increased according to increasing of time and concentration. These results suggest that cisplatin, heptaplatin and sunpla are a novel anticancer agent against human melanoma cell.l.

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.

DNA Ploidy in Anaplastic Carcinoma of the Thyroid Gland by Image Analysis (갑상선 역형성암종의 DNA 배수성에 관한 화상분석학적 연구)

  • Lee, Ji-Shin;Lee, Min-Cheol;Park, Chang-Soo;Juhng, Sang-Woo
    • The Korean Journal of Cytopathology
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    • v.6 no.1
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    • pp.10-17
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    • 1995
  • Anaplastic carcinoma of the thyroid gland is one of the most malignant tumors. Recently, DNA ploidy measured by flow cytometry and image analysis has been suggested as an additional useful indicator of tumor behavior. Studies on the occurrence and clinical significance of DNA aneuploidy in anaplastic carcinoma of the thyroid are rare. In this study, the pattern of DNA ploidy was measured by image analysis on Papanicolaou stained slides in four cases of anaplastic carcinoma and also measured by flow cytometry using paraffin blocks in two cases. In all cases of anaplastic carcinoma, DNA aneuploidy was found by image analaysis. By flow cytometry, one case had a diploid peak and the other case had an aneuploid peak. According to the above results, we conclude that anaplastic carcinoma of the thyroid glands have a high incidence of DNA aneuploidy and image analysis using Papanicolaou stained slides is a useful method in detecting DNA aneuploidy.

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