• 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.

• 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.

• 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.

• 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.

• 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.

• The Korean Journal of Malacology
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• v.28 no.1
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• pp.37-43
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• 2012

In this study, we invesgated a cytogenetic analysis of the Pacific triploid abalone, Haliotis discus hannai induced by low temperature treatment. We got a lot of mitotic metaphase chromosome spreads from the triploid and diploid Pacific abalones' hatched larvae (trochophores). The chromosome number of diploid abalone was 2n = 36 and that of triploid abalone was 3n = 54, so the chromosome number of triploid abalone was 1.5 times higher than that of diploid abalone. We developed a modified flow cytometric method for Pacific abalone from the existing methods. We uesd 51 months aged triploid and diploid Pacific abalones' hemolymph to get the DNA contents by flow cytometry. The DNA content of diploid abalone was 1.743 pg/cell and the DNA content of triploid abalone was 1.49 times higher than that of diploid one. It proved that triploid abalone was consisted with two sets of maternal diploid and one set of paternal genome.

• Parasites, Hosts and Diseases
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• v.49 no.3
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• pp.229-234
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• 2011

In order to assess changes in the activity of immunecompetency present in Crassostrea gigas infected with Marteilioides chungmuensis (Protozoa), the total hemocyte counts (THC), hemocyte populations, hemocyte viability, and phagocytosis rate were measured in oysters using flow cytometry. THC were increased significantly in oysters infected with M. chungmuensis relative to the healthy appearing oysters (HAO) (P<0.05). Among the total hemocyte composition, granulocyte levels were significantly increased in infected oysters as compared with HAO (P<0.05). In addition, the hyalinocyte was reduced significantly (P<0.05). The hemocyte viability did not differ between infected oysters and HAO. However, the phagocytosis rate was significantly higher in infected oysters relative to HAO (P<0.05). The measurement of alterations in the activity of immunecompetency in oysters, which was conducted via flow cytometry in this study, might be a useful biomarker of the defense system for evaluating the effects of ovarian parasites of C. gigas.

• Biotechnology and Bioprocess Engineering:BBE
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• v.8 no.2
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• pp.64-75
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• 2003

Aptamers are functional nucleic acids that can specially bind to proteins, peptides, amino acids. nucleotides, drugs, vitamins and other organic and inorganic compounds. The aptamers are identified from random DNA or RNA libraries by a SELEX (systematic evolution of ligands by exponential amplification) process. As aptamers have the advantage, and potential ability to be released from the limitations of antibodies, they are attractive to a wide range of therapeutic and diagnostic applications. Aptamers, with a high-affinity and specificity, could fulfil molecular the recognition needs of various fields in biotechnology. In this work, we reviewed some aptamer Selection techniques, properties, medical applications of their molecules and their biotechnological applications, such as ELONA (enzyme linked oligonucleotide assay), flow cytometry, biosensors, electrophoresis, chromatography and microarrays.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.43 no.2
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• pp.128-133
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• 1998

Analyses of now karyotypes using different maize (Zea mays L.) inbred lines have been performed. The accumulation and isolation of high quality and quantity metaphase chromosomes from root tips can be achieved from many kinds of maize lines. The chromosome suspensions were prepared by a simple slicing method from synchronized maize root tips and analyzed with a now cytometry. The variations of experimental now karyotypes were detected among inbred lines in terms of the positions and/or the numbers of chromosome peaks. The 2C DNA amount among 8 inbred lines ranged from 5.09 to 5.52 pg. The variability of DNA content in maize chromosome 1 was 9.1 % ranging from 0.685 to 0.747 pg. The selection of appropriate maize lines is critical for sorting specific single chromosome types. At least five different chromosome types can be discriminated and sorted from five maize lines.

• IMMUNE NETWORK
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• v.5 no.2
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• pp.105-109
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• 2005

Background: Dendritic cells (DCs) are the most potent APCs (antigen-presenting cells) and playa critical role in immune responses. Galectin-3 is a biological lectin with a beta-galactoside binding affinity. Recently, proteomic analysis revealed the presence of galectin-3 in the exosome of mature DCs. However, the expression and function of galectin-3 in DCs remains unclear yet. Methods: We used bone marrow-derived DCs of mouse and showed the expression of galectin-3 in DCs by using flow cytometry analysis and Western blot analysis. Results: Galectin-3 was determined as single band of 35 kDa in Western blot analysis. Flow cytometry analysis showed the major growth factor for DCs, granulocyte-macrophage colony stimulating factor (GM-CSF) and maturing agents, anti-CD40 monoclonal antibody (mAb) and lipopolysaccharide (LPS) consistently increased the intracellular expression of galectin-3 in DCs compared to medium alone. In addition, DCs treated with maturing agents did marginally express galectin-3 on their surface. Conclusion: This study suggests that galectin-3 in DCs may be regulated by critical factors for DC function.