• Title/Summary/Keyword: flow cytometry

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Establishment of the cell lines with plant regeneration ability and low ploidy level in Dianthus acicularis with the aid of flow cytometry analysis

  • Shiba, Tomonori;Mii, Masahiro
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2005.11a
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    • pp.112-119
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    • 2005
  • Efficient plant regenerationsystem from cell suspension cultures was established in D. acicularis (2n = 90) by monitoring ploidy level and visual selection of the cultures. The highly regenerable cell lines selected maintained original ploidy level and consisted of compact cell clumps with yellowish color and relatively moderate growth, suggesting that it is possible to select visually the highly regenerable cell lines with the original ploidy level. All the regenerated plantlets from the highly regenerable cell cultures exhibited normal phenotypes and no variations in ploidy level were observed by flow cytometry (FCM) analysis.

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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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Flow Cytometric Analysis of Apoptosis Inhibition by Silkworm Hemolymph

  • Lee, Won-Jong;Kim, Eun-Jeong;Park, Tae-Hyeon
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.151-154
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    • 2000
  • The effect of silkworm hemolymph on insect cell apoptosis was investigated. The addition of silkworm hemolymph into the culture medium either before or during the baculovirus infection increased the host cell longevity. Silkworm hemolymph also inhibits apoptosis induced by actinomycin D, the RNA synthesis inhibitor. In this study, a flow cytometry was used for the quantitative analysis of apoptosis inhibition by silkworm hemolymph.

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Cytogenetic Analysis of Induced Hybrid between Common Carp (Cyprinus carpio) and Crucian Carp (Carassius auratus) (잉어(Cyprinus carpio)와 붕어(Carassius auratus)간 잡종의 세포유전학적 분석)

  • 남윤권;오승용;조재윤;김동수
    • Journal of Aquaculture
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    • v.11 no.1
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    • pp.77-81
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    • 1998
  • Cytogenetic analyses were carried out with induced hybrid between common carp (Cyprinus carpio) female and crucian carp (Carassius auratus) male. The erythrocytic measurement revealed that cellular and nuclear size of induced hybrids were intermediate between those of paremtal species. The modal chromosome numbers of common carp, crucian carp and its hybid were same as 2n=100. The DNA content of induced hybrids determined based on flow cytometry was 3.7pg/cell which corresponding to intermediate value between the carp (3.6ph/cell) and crucian carp (3.8pg/cell)

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Characterization of Cultured Angelica gigas Microspores by Flow Cytometry (당귀 배양 소포자의 Flow Cytometric 특성)

  • Park, Chung-Heon;Seong, Nak-Sul;Yu, Hong-Seob;Pauls, K. Peter
    • Korean Journal of Medicinal Crop Science
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    • v.5 no.3
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    • pp.196-201
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    • 1997
  • To characterize active cells during microspore culture of Angelica gigas, flow cytometric and epifluorescent techniques were applied. The knowledge obtained from these types of studies will give us insight into early stage in plant development and may lead to the application of microspore-derived from haploid plants for breeding in recalcitrant species. Viability of cultured microspore differed depending on the developmental stages. Frequencies of active cells from tetrad, uni-nucleate, bi-nucleate and matured pollen were 12.8, 49.3, 42.3 and 31.7%, respectively. Alive microspores have luminescent the green fluorescence stained with FDA and blue fluorescence stained with DAPI.

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Plant genome analysis using flow cytometry

  • Lee Jai-Heon;Kim Kee-Young;Chung Dae-Soo;Chung Won Bok;Kwon Oh-Chang
    • Proceedings of the Korean Society of Crop Science Conference
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    • 1999.05a
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    • pp.162-163
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    • 1999
  • The goal of this research was (1) to describe the conditions and parameters required for the cell cycle synchronization and the accumulation of large number of metaphase cells in maize and other cereal root tips, (2) to isolate intact metaphase chromosomes from root tips suitable for characterization by flow cytometry, and (3) to construct chromosome-specific libraries from maize. Plant metaphase chromosomes have been successfully synchronized and isolated from many cereal root-tips. DNA synthesis inhibitor (hydroxyurea) was used to synchronize cell cycle, follwed by treatement with trifluralin to accumulate metaphase chromosomes. Maize flow karyotypes show substantial variation among inbred lines. thish variation should be sueful in isolating individual chromosome types. In addition, flow cytometry is a useful method to measure DNA content of individual chromosomes in a genotyps, and to detect chromosomal variations. Individual chromosome peaks have been sorted from the maize hybrid B73/Mol7. Libraries were generated form the DOP-PCR amplification product from each peak. To date, we have analyzed clones from a library constructed from the maize chromosome 1 peak. Hybridization of labeled genomic DNA to clone inserts indicated that $24\%,\;18\%,\;and\;58\%$ of the clones were highly repetitive, medium repetitive, and low copy, respectively. Fifty percent of putative low cpoy clones showed single bands on inbred screening, blots, and the remaining $50\%$ were low copy repeats. Single copy clones showing polymorphism will be mapped using recombinant inbred mapping populations. Repetitive clones are being characterized by Southern blot analysis, and will be screened by in situ hybridization for their potential utility as chromosome specific markers.

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FLOW CYTOMETRIC ANALYSIS OF LYMPHOCYTES IN NORMAL AND INFLAMED PULP (유세포분석기를 이용한 정상치수조직과 염증성 치수조직 내의 임파구 분포에 관한 연구)

  • Kim, Seon-Ah;Bae, Kwang-Shik;Im, Seong-Sam
    • Restorative Dentistry and Endodontics
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    • v.22 no.1
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    • pp.374-387
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    • 1997
  • The purpose of this study was to examine the distribution of lymphocyte populations in normal, reversibly inflamed and irreversibly inflamed human dental pulp tissues using flow cytometry. Flow cytometry, with specific antibody and fluorochrome reagent allows us to know cellular properties of hematolymphoid cells by measuring fluorescence of stained cells. Before extirpation of pulps in routine endodontic treatment, the clinical diagnosis were performed by symptom. The extirpated pulp tissues were divided into normal pulp group (N=5), reversible pulpit is group(N=10) and irreversible pulpitis group(N=7). The specimen was placed into RPMI 1640 medium, minced into small pieces, and then digested in medium with collagenase. The cell suspension was resuspended in PBS for monoclonal antibody staining of T lymhocytes(CD3+), B lymphocytes (CD19+), T helper cell (CD4+) and T supressor cell (CD8+). The percentages of cells were counted by FACStar(BD) flow cytometer. Following results were obtained; 1. In the most normal and inflamed pulps, the percentages of T lymphocyte, B lymphocytes, T helper cell and T suppressor/cytotoxic cell were less than 1 % in total counted pulpal cells. 2. The higher percentages of T, B, T helper and T suppressor cells were observed in irreversible pulpitis group as compared with the normal pulp and reversible pulpitis group but the differences between groups were not statistically significant (p>0.05). 3. The percentages of T helper cells (CD4 + cells) were greater than that of T suppressor/cytotoxic cells (CD8 + cells) in the inflamed pulps.

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Expression of the red sea bream iridovirus (RSIV) capsid protein using a yeast surface display method (효모표면표출(YSD) 기법을 이용한 참돔 이리도바이러스(RSIV) 외피단백질의 발현)

  • Suh, Sung-Suk;Park, Mirye;Hwang, Jinik;Lee, Taek-Kyun
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.15 no.8
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    • pp.5412-5418
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    • 2014
  • The red seabream iridovirus (RSIV), which belongs to the iridoviridae, causes infectious fish diseases in many Asian countries, leading to considerable economic losses to the aquaculture industry. Using the yeast surface display (YSD) technique, a new experimental system was recently developed for the detection and identification of a variety of marine viruses. In this study, a coat protein gene of RSIV was synthesized based on the nucleotide sequence database and subcloned into the yeast expression vector, pCTCON2. The expression of viral coat proteins in the yeast strain, EBY100, was detected by flow cytometry and Western blot analysis. Finally, they were isolated from the yeast surface through a treatment with ${\beta}$-mercaptoethanol. The data suggests that the YSD system can be a useful method for acquiring coating proteins of marine viruses.

Analysis of Membrane Integrity and Mitochondrial Activity in Fresh and Cryopreserved Boar Sperm Using Flow Cytometry

  • Park C. S.;Li Z. H.;Sung N. D.;Jin D. I.;Cong P. Q.;Kim E. S.;Song E. S.;Yi Y. J.
    • Reproductive and Developmental Biology
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    • v.29 no.4
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    • pp.253-257
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    • 2005
  • This study was carried out to evaluate the effects of washing medium, breed and washing temperature of fresh and frozen-thawed boar sperm on mitochondrial activity and membrane integrity by flow cytometry. More than $80\%$ of fresh sperm washed with mTLP-PVA medium at $20^{\circ}C$ exhibited an intact membrane and a functional mitochondrion. With frozen-thawed samples, a large number of sperm showed both damaged membrane $(36.4\~46.9\%)$ and nonfunctional mitochondrion $(55.1\~71.1\%)$ in the mTLP-PVA and BTS washing media at $20^{\circ}C$. There were no breed effects of fresh and frozen-thawed sperm on mitochondrial activity and membrane integrity. The percentages of damaged membrane of fresh and frozen sperm, respectively, were higher at $4^{\circ}C$ washing temperature than at $20^{\circ}C$ washing temperature in the mTLP-PVA medium. We found that washing medium and washing temperature of fresh and frozen-thawed boar sperm were important for the analyses of mitochondrial activity and membrane integrity by flow cytometry.

Evaluation of Sperm Sex-Sorting Method using Flow Cytometry in Hanwoo (Korean Native Cattle)

  • Yoo, Han-Jun;Lee, Kyung-Jin;Lee, Yong-Seung;Lee, Chang-Woo;Park, Joung-Jun;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Journal of Embryo Transfer
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    • v.27 no.1
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    • pp.37-43
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    • 2012
  • This study evaluated a method of sorting X and Y chromosomes based on size using the forward angle light scatter related refractive index (FSC) of a flow cytometer. Hanwoo bulls sperm were separated to X and Y chromosomes by the parameters of FSC or Hoechst 33342 intensity. As a result, using monitor program linked flow cytometry during sorting processing, the purities were $97{\pm}0.57$ or $96{\pm}0.67%$ for the X-fraction and $96{\pm}0.33$ or $97{\pm}1.33%$ for the Y-fraction in the two sperm sorting methods. There were no differences in the X and Y ratios (X and Y %) between the sperm sorting methods based on FSC or DNA content. The proportions of female and male embryos used for in vitro fertilization and development were $66.03{\pm}3.31$ or $69.37{\pm}1.41%$, and $70.56{\pm}2.42$ or $56.11{\pm}3.09%$ when sperm were processed using the sex sorting method by FSC or Hoechst 33342. In conclusion, further study is needed to determine the optimum procedure and improve the nozzle to enhancing sorting accuracy or efficiency. Also, the findings of this study do not negate the possibility that the difference method of sperm sorting cannot use a UV laser beam.