• Title/Summary/Keyword: Flow cytometric

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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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Comparison of Flow Cytometric and Morphological Assessment on Viability of Spermatozoa during Freezing Process of Boar Semen (돼지 정액의 동결과정중 정자 생존성에 대한 Flow Cytometric 및 형태학적 평가의 비교)

  • ;;L.A. Johnson
    • Journal of Embryo Transfer
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    • v.14 no.1
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    • pp.69-77
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    • 1999
  • 본 연구는 돼지 정액의 동결과정동안 flow cytometric 분석에 의한 정액내 생존정자의 비율을 조사하여 주관적으로 평가되는 활력 및 정상첨체율(normal apical ridge ; NAR)과 비교하여 정자의 손상과 생존성에 대한 적절한 평가법을 찾기 위하여 실시하였다. 동결과정 중 정액채취, 냉각, 예비동결 및 동결융해 후에 flow cytometric 분석에 의한 정자 생존율은 각각 93.0$\pm$3.6, 85.1$\pm$3.9, 28.9$\pm$6.8 및 26.1$\pm$5.9%이었다. 동결처리동안에 생존율은 예비동결 및 동결융해 후 가장 많은 정자사멸로 동결상태 이전의 생존율보다 유의적으로 낮게 나타났다. (p<0.05). 평가기법으로 정액 채취시 활력, NAR율 및 생존율을 조사한 결과 각각 91.0$\pm$4.2, 96.8$\pm$2.5 및 92.2$\pm$3.2%로 NAR율이 생존성 및 활력보다 높게 평가되었으며, 생존율이 활력보다 다소 높게 평가되었다. 그러나 동결융해 후에는 각각 44.0$\pm$8.9, 49.0$\pm$7.9 및 35.6$\pm$9.7%로 활력이 생존율보다 다소 높게 평가되었다. 전체적으로 NAR율은 활력은 생존율보다 높게 평가되었으며, SYBR-14 / PI(propidium iodide) 이중형광염색법에 의한 flow cytometric 평가법으로 생존율은 동결되지 않은 정액에서의 활력 및 NAR 평가보다 다소 민감하게 나타났다. 이러한 결과로 미루어보아 SYBR-14 / PI 형광염색에 의한 flow cytometry의 생존성 평가는 동결되지 않은 정액의 평가방법으로는 적절하지만 동결된 정액의 생존성 평가는 부적절한 것으로 사료되었다.

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A Rapid Assessing Method of Drug Susceptibility Using Flow Cytometry for Mycobacterium tuberculosis Isolates Resistant to Isoniazid, Rifampin, and Ethambutol

  • Lee, Sun-Kyoung;Baek, Seung-Hun;Hong, Min-Sun;Lee, Jong-Seok;Cho, Eun-Jin;Lee, Ji-Im;Cho, Sang-Nae;Eum, Seok-Yong
    • Tuberculosis and Respiratory Diseases
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    • v.85 no.3
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    • pp.264-272
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    • 2022
  • Background: The current conventional drug susceptibility test (DST) for Mycobacterium tuberculosis (Mtb) takes several weeks of incubation to obtain results. As a rapid method, molecular DST requires only a few days to get the results but does not fully cover the phenotypic resistance. A new rapid method based on the ability of viable Mtb bacilli to hydrolyze fluorescein diacetate to free fluorescein with detection of fluorescent mycobacteria by flow cytometric analysis, was recently developed. Methods: To evaluate this cytometric method, we tested 39 clinical isolates which were susceptible or resistant to isoniazid (INH) or rifampin (RIF), or ethambutol (EMB) by phenotypic or molecular DST methods and compared the results. Results: The susceptibility was determined by measuring the viability rate of Mtb and all the isolates which were tested with INH, RIF, and EMB showed susceptibility results concordant with those by the phenotypic solid and liquid media methods. The isolates having no mutations in the molecular DST but resistance in the conventional phenotypic DST were also resistant in this cytometric method. These results suggest that the flow cytometric DST method is faster than conventional agar phenotypic DST and may complement the results of molecular DST. Conclusion: In conclusion, the cytometric method could provide quick and more accurate information that would help clinicians to choose more effective drugs.

Flow Cytometric Analysis of Human Lysozyme Production in Recombinant Saccharomyces cerevisiae

  • Peterson Marvin S.;Kim Myoung-Dong;Han Ki-Cheol;Kim Ji-Hyun;Seo Jin-Ho
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.7 no.1
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    • pp.52-55
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    • 2002
  • Flow cytometric techniques were used to investigate cell size, protein content and cell cycle behavior of recombinant Saccharomyces cerevisiae strains producing human lysozyme (HLZ). Two different signal sequences, the native yeast $MF\alpha1$ signal sequence and the rat $\alpha-amylase$ signal sequence, were used for secretion of HLZ. The strain containing the rat $\alpha-amylase$ signal sequence showed a higher level of internal lysozyme and lower specific growth rates. Flow cytometric analysis of the total protein content and cell size showed the strain harboring the native yeast signal sequence had a higher total protein content than the strain containing the rat $\alpha-amylase$ signal sequence. Cell cycle analysis indicated that the two lysozyme producing recombinant strains had an increased number of cells in the $G_2+M$ phase of the yeast cell cycle compared with the host strain SEY2102.

Flow Cytometric Analysis of the Effect of Silkworm Hemolymph on the Baculovirus-Induced Insect Cell Apoptosis

  • Rhee, Won-Jong;Park, Tai-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.11 no.5
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    • pp.853-857
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    • 2001
  • The effect of silkworm hemolymph on the inhibition of baculovirus-induced insect cell apoptosis was quantitatively investigated using a flow cytometric analysis. Spodoptera frugiperda (Sf9) cell and Autographa californica nuclear polyhedrous virus (AcNPV) were used as a model for insect cell and baculovirus in this study, respectively. Compared with a mammalian cell cycle, the fraction of G1 cells was relatively small in the Sf9 cell cycle. Silkworm hemolymph did not affect the Sf9 cell-cycle distribution before the baculovirus infection. However, the fraction of cells which are not in the sub-G1 phase remained at a high level for 3 days after the infection in the medium without silkworm hemolymph, while it remained at a high level for 7 days after the infection in the medium supplemented with silkworm hemolymph. The fractions of apoptotic cells in the sub-G1 phase were $4.7\%$, and 4 days after infection, $22.7\%$, in the media with and without silkworm hemolymph, respectively.

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Flow cytometry As a Tool for Monitoring Immune Parameters of the Manila clam Ruditapes philippinarum

  • Park, Kyung-Il;Philippe Soudant;Park, Kwang-Sik
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2003.05a
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    • pp.369-369
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    • 2003
  • Hemocytes in marine bivalves play important immunological roles in discrimination, opsonization and phagocytosis of foreign materials as a defense mechanism. In this study we report the flow cytometric implications to investigate the immune parameters such as the compositional and the functional characteristics of hemocytes isolated from the Manila clams, Ruditapes philippinarum. Heterogeneity of the hemocytic cell population was determined by the forward scatter (FSC) and side scatter (SSC) cytometric profile which showed three populations: granulocytes, hyalinocytes and small agranular cells. In addition, phagocytosis rate was measured after adding fluorescent-labeled particles. The data were initially analysed for two-parameters: FSC and SSC, then the fluorescent (FL 1) frequency distribution histogram of the hemocyte population was subsequently obtained.

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Prognostic significance of minimal residual disease detected by a simplified flow cytometric assay during remission induction chemotherapy in children with acute lymphoblastic leukemia

  • Koh, Kyung-Nam;Park, Mee-Rim;Kim, Bo-Eun;Im, Ho-Joon;Park, Chan-Jeoung;Jang, Seong-Soo;Chi, Hyun-Sook;Seo, Jong-Jin
    • Clinical and Experimental Pediatrics
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    • v.53 no.11
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    • pp.957-964
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    • 2010
  • Purpose: Our study attempted to determine the prognostic significance of minimal residual disease (MRD) detected by a simplified flow cytometric assay during induction chemotherapy in children with B-cell acute lymphoblastic leukemia (B-ALL). Methods: A total of 98 patients were newly diagnosed with precursor B-ALL from June 2004 to December 2008 at the Asan Medical Center (Seoul, Korea). Of those, 37 were eligible for flow cytometric MRD study analysis on day 14 of their induction treatment. The flow cytometric MRD assay was based on the expression intensity of CD19/CD10/CD34 or aberrant expression of myeloid antigens by bone marrow nucleated cells. Results: Thirty-five patients (94.6%) had CD19-positive leukemic cells that also expressed CD10 and/or CD34, and 18 (48.6%) had leukemic cells with aberrant expression of myeloid antigens. Seven patients with ${\geq}1%$ leukemic cells on day 14 had a significantly lower relapse-free survival (RFS) compared to the 30 patients with lower levels (42.9 % [18.7%] vs. 92.0% [5.4%], $P$=0.004). Stratification into 3 MRD groups (${\geq}1%$, 0.1-1%, and <0.1%) also showed a statistically significant difference in RFS (42.9% [18.7%] vs. 86.9% [8.7%] vs. 100%, $P$=0.013). However, the MRD status had no significant influence on overall survival. Multivariate analysis demonstrated that the MRD level on day 14 was an independent prognostic factor with borderline significance. Conclusion: An MRD assay using simplified flow cytometry during induction chemotherapy may help to identify patients with B-ALL who have an excellent outcome and patients who are at higher risk for relapse.

Comparison of polymerase chain reaction for antigen receptor gene rearrangement and flow cytometric analysis for the diagnosis of canine lymphoma

  • Song, Ru-Hui;Yu, Do-Hyeon;Kim, Jun-Hwan;Lee, Hyun-Seok;Lee, Da-Mi;Park, Chul;Yu, Il-Jung;Park, Jin-Ho
    • Korean Journal of Veterinary Service
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    • v.34 no.3
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    • pp.265-271
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    • 2011
  • Lymphoma is the most common hematopoietic malignancy in dogs. Diagnosis of lymphoma is classically performed by morphological assessment and immunohistochemistry. But some cases in the early stage are difficult to distinguish and need more objective and accurate methods. So, Polymerase chain reaction (PCR) for antigen receptor rearrangements (PARR) and flow cytometric immunophenotype of lymphoma have been developed continuously. In this study, we performed these two methods to classify lymphoma type in 3 cases. According to PARR analysis, B cell origin lymphoma was diagnosed in two of three cases by testing PBMC and lymph node. All fine needle aspiration (FNA) samples of lymph nodes had high expression of CD21 on >88% of total cell population and PBMC samples also showed high expression of CD21 on >30% of total lymphocytes in those two cases, while the expression of CD3, CD4 and CD8 was absent. These results suggest that concurrent use of PARR and flow cytometric immunophenotype is more effective and valuable tool for the diagnosis and monitoring of canine lymphoma patients.

Flow cytometric immunophenotyping of canine adipose-derived mesenchymal stem cells (ADMSCs) and feline ADMSCs using anti-human antibodies

  • Ko, Minho;Lee, Kwon Young;Kim, Sae Hoon;Kim, Manho;Choi, Jung Hoon;Im, Wooseok;Chung, Jin Young
    • Korean Journal of Veterinary Research
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    • v.58 no.1
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    • pp.33-37
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    • 2018
  • Various trials have been conducted to develop therapies for serious untreatable diseases. Among these, those using stem cells have shown great promise, and adipose-derived mesenchymal stem cells (ADMSCs) are easier to obtain than other types of stem cells. Prior to clinical trials, characterization of ADMSCs with monoclonal antibodies should be performed. However, it is difficult to use species-specific antibodies for veterinarians. This study was conducted to confirm the panel of human antibodies applicable for use in immunophenotypic characterization of canine adipose-derived stem cells and feline ADMSCs extracted from subcutaneous adipose tissue collected during ovariohysterectomy. For flow cytometric immunophenotyping, the third passages of canine ADMSC and feline ADMSC and human CD31, CD34, CD42, CD44, CD62 and CD133 antibodies were used. Of these, CD133 reacted with canine cells (3.74%) and feline cells (1.34%). CD133 is known as a marker related with more primitive stem cell phenotype than other CD series. Because this human CD133 was not a species-specific antibody, accurate percentages of immunoreactivity were not confirmed. Nevertheless, the results of this study confirmed human CD133 as a meaningful marker in canine and feline ADMSCs.