• Title/Summary/Keyword: flow cytometry

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Design of a customizable fluorescence detection system for fluorescently labeled tumor cells (형광 발현 암세포 탐지를 위한 맞춤형 검출시스템 개발)

  • Cho, Kyoungrae;Seo, Jeong-hyeok;Choe, Se-woon
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.23 no.3
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    • pp.261-266
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    • 2019
  • Flow cytometry is an electrical detection system that provides precise and diverse optical properties to cells and micro particles. Flow cytometry, which provides multidimensional information including cell size and granularity through light scattering and fluorescence emission generated by the induction of light of a specific wavelength to the fluorescently treated cells or micro particles, plays an important role in biomedical and biophysical fields. However, it has some drawbacks such as high cost, size of the instrument and limitation in selecting fluorescent dyes. Therefore, in this paper, a low cost compact fluorescent detection system is developed using light-emitting diode and microcontroller. The proposed fluorescence detection system has a replaceable the light source/fluorescence filter/photodetector and constructed by 3D printer, so that the user can design a customized system according to the selected fluorescent dyes. The fluorescence intensity was measured by varying the number of fluorescently labeled cells, and the measured intensities showed a high linearity within the tested concentration ranges.

Protective Effects on Gastric Lesion of Ursolic acid (Ursolic acid의 위 손상에 대한 방어 효과)

  • Kim, Sun Whoe;Hwang, In Young;Lee, Sun Yi;Jeong, Choon Sik
    • Journal of Food Hygiene and Safety
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    • v.31 no.4
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    • pp.286-293
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    • 2016
  • This study is an experiment for gastric protective effects of ursolic acid. In order to identify the effects of ursolic acid on gastrointestinal disorder, acute and chronic gastritis were also observed using HCl ethanol and indomethacin-induced gastric lesion models, respectively. As for gastric acid, it was also identified through proton pump ($H^+/K^+-ATPase$) inhibiting activity. In regards to protective factor for gastric damage, prostaglandin $E_2$ ($PGE_2$) was quantitatively analyzed. Antibacterial activity experiment was done on Helicobacter pylori (H.pylori), which is known to be the causing factor of chronic gastritis, gastric ulcer and gastric cancer. By making use of AGS cell, it was confirmed that ursolic acid was involved in apoptosis of gastric cancer cell through 4',6-diamidino-2-phenylindol (DAPI) staining and flow cytometry analysis. As a result, ursolic acid reduced gastric lesions caused by HCl ethanol and indomethacin. Ursolic acid inhibited acid secretion by inhibiting proton pump ($H^+/K^+-ATPase$), which is the gastric acid secreting enzyme involved at the final phase of gastric acid secretion. And ursolic acid was identified with gastric mucosa protection effects by increasing the concentration of $PGE_2$, a protective factor of gastric mucosa preservation. The antibacterial activity on H. pylori, which is aggressive factor in gastrointestinal disorder, ursolic acid showed inhibitory effects on H. pylori colonization. In the DAPI nuclear staining, unlike the control group, shape of the nucleus has deformed, and has been observed either shrinked cell or chromatin condensation phenomenon. In the Flow cytometry assay, confirmed the growth rate of apoptosis in a concentration-dependent manner.

Effect of Simvastatin collagen graft on wound healing of defective bone (Simvastatin이 골결손부 회복에 미치는 영향)

  • Kang, Jung-Ho;Kim, Gyu-Tae;Choi, Yong-Suk;Lee, Hyeon-Woo;Hwang, Eui-Hwan
    • Imaging Science in Dentistry
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    • v.38 no.3
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    • pp.133-146
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    • 2008
  • Purpose : To observe and evaluate the effects of Simvastatin-induced osteogenesis on the wound healing of defective bone. Materials and Methods : 64 defective bones were created in the parietal bone of 32 New Zealand White rabbits. The defects were grafted with collagen matrix carriers mixed with Simvastatin solution in the experimental group of 16 rabbits and with collagen matrix carriers mixed with water in the controlled group. The rabbits were terminated at an interval of 3, 5, 7, and 9 days, 2, 4, 6, and 8 weeks after the formation of defective bone. The wound healing was evaluated by soft X-ray radiography. The tissues within defective bones were evaluated through the analysis of flow cytometry for the manifestation of Runx2 and Osteocalcin, and observed histopathologically by using H-E stain and Masson's trichrome stain. Results : 1. In the experimental group, flow cytometry revealed more manifestation of Runx2 at 5, 7, and 9 days and Osteocalcin at 2 weeks than in the controlled groups, but there was few difference in comparison with the controlled group. 2. In the experimental group, flow cytometry revealed considerably more cells and erythrocytes at 5, 7, and 9 days in comparison with the controlled group. 3. In the experimental group, soft x-ray radiography revealed the extended formation of trabeculation at 2, 4, 6, and 8 weeks. 4. Histopathological features of the experimental group showed more fibroblasts and newly formed vessels at 5 and 7 days, and the formation of osteoid tissues at 9 days, and the newly formed trabeculations at 4 and 6 weeks. Conclusion : As the induced osteogenesis by Simvastatin, there was few contrast of the manifestation between Runx2 and Osteocalcin based on the differentiation of osteoblasts. But it was considered that the more formation of cells and erythrocytes depending on newly formed vessels in the experimental group obviously had an effect on the bone regeneration.

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Genome size of 15 Lamiaceae taxa in Korea (한국산 꿀풀과 15 분류군에 대한 유전체양 조사)

  • Lee, Yoonkyung;Kim, Sangtae
    • Korean Journal of Plant Taxonomy
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    • v.47 no.2
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    • pp.161-169
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    • 2017
  • The genome size is one of the basic characters of an organism, and it is widely applied in various fields of biology, such as systematics, breeding biology, population biology, and evolutionary biology. This factor was recently highlighted in genome studies because choosing a representative of a plant group having the smallest genome size is important for the efficiency of a genome project. For the estimation of the genome size, flow cytometry has recently been highlighted because it is a convenient, fast, and reliable method. In this study, we report the genome sizes of 15 taxa of Lamiaceae from nine genera distributed in Korea using flow cytometry. Data pertaining to the genome size for all of our species have not been reported thus far, and the data from Agastache, Clinopodium, Elsholtzia, and Isodon are the first reported for each genus. The genome sizes of 15 genera and 39 species were reported to the Plant DNA C-values Database (http://data.kew.org/cvalues/). Scutellaria indica L. has a genome size of 0.37 pg (1C). This is the fourth smallest value among the 98 Lamiaceae taxa in the Angiosperm DNA C-value Database, indicating that this taxon can be used as a reference species in the genome studies in Lamiaceae as a native Korean species. The largest genome size observed in this study is in Phlomis umbrosa Turcz. (1C=2.60 pg), representing the possible polyploidy origin of this species in the family.

Analysis of Proliferative Potentials in Meningiomas by Ki-67, Proliferating Cell Nuclear Antigen, and Flow Cytometry (Ki-67, Proliferating Cell Nuclear Antigen, Flow Cytometry를 이용한 수막종의 증식력 분석)

  • Ahn, Jae Sung;Kim, Jeong Hoon;Kwun, Byung Duk
    • Journal of Korean Neurosurgical Society
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    • v.30 no.7
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    • pp.861-869
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    • 2001
  • Objective : In this study, we investigated the relationship between the histologic grading of meningiomas and proliferative potentials determined by the Ki-67, proliferating cell nuclear antigen(PCNA) and flow cytometry (FCM) with the aim of determining whether these potentials can be used as a parameter to the proliferative activity, in particular of atypical and malignant meningiomas. Methods : This study consisted of 47 meningiomas(6 malignant, 14 atypical, and random sampled 27 benign meningiomas). By immunohistochemical staining of Ki-67 and PCNA on formalin-fixed, paraffin-embedded sections, the anti-human rabbit polyclonal antibody against Ki-67 antigen and anti-PCNA monoclonal antibody(PC10) scores were counted. FCM was also performed on paraffin-embedded tissue using a selective staining technique for DNA. DNA ploidy, S-phase fraction, and proliferative index(PI)) were determined. Results : The results are summarized as follows ; 1) Proliferation rates as assessed by Ki-67 and PCNA closely correlated with the degree of anaplastic histologic features. 2) Proliferative potentials determined by FCM(S-phase fraction and PI) were not able to distinguish between benign and atypical/malignant meningiomas. 3) DNA ploidy was not a useful indicator of histologic grade in these tumors. 4) Proliferative potentials such as Ki-67 staining index(SI) and PCNA SI did not correlate with the ploidy pattern. 5) There was a linear correlation between Ki-67 SI and PCNA SI, but we could not find a correlation between Ki-67 SI and S-phase fraction or PI. Our results also did not show a statistically signficant correlation between PCNA SI and S-phse fraction or PI. Conclusions : We conclude that evaluation of the proliferative potentials with Ki-67 and PCNA is important as an additional factor for the prediction of malignancy in meningiomas. A dual study of Ki-67 and PCNA SIs on the same tissue might improve the accuracy with which the proliferative potential of a tumor can be predicted. We demonstrated that FCM in meningiomas is not valuable in predicting the behavior of these neoplasms, but we did observe a trend toward more malignancy with higher percent S-phase fraction and higher PI. Analysis of the S-phase fraction and PI might therefore be a useful tool to discriminate among histologic grades of meningiomas.

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Assessment of Immune Parameters of the Wild Pacific Oyster (Crassostrea gigas) using a Flow Cytometry and Neutral Red Retention Assay (유세포 분석기와 Neutral Red Retention Assay를 이용한 자연산 참굴(Crassostrea gigas)의 면역력 측정)

  • Hong, Hyun-Ki;Kang, Hyun-Sil;Kim, Young-Ok;Choi, Kwang-Sik
    • Ocean and Polar Research
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    • v.34 no.2
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    • pp.137-149
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    • 2012
  • Hemocyte parameters of the wild Pacific oyster Crassostrea gigas inhabiting intertidal zones in small bays (Gwangyang and Jinhae Bay) on the southern coast of Korea were evaluated using flow cytometry and neutral red retention (NRR) assay. Morphological features, cell count, mortality, DNA damage, phagocytosis, and lysosomal membrane stability of hemocytes were analyzed. Three types of hemocytes were identified in the oyster hemolymph: granulocytes, hyalinocytes, and blast-like cells. Immune related functions of hemocyte including phagocytosis and lysosomal membrane stability were significantly different among the study areas (P<0.05), while cell count, mortality, and DNA damage of hemocytes were not significantly different. In Gwangyang Bay, phagocytosis of granulocytes and lysosomal membrane stability of oyster hemocytes inhabiting inside bay were significantly lower than those of oyster hemocytes in outside bay (P<0.05), indicating that oysters in inside bay of Gwangyang were relatively suppressed the immunological function in hemocytes. Contrary to Gwangyang Bay, immune parameters of oyster hemocytes in Jinhae Bay not showed the difference between sampling sites. In conclusion, flow cytometry and NRR assay using oyster hemocyte has a powerful tool to investigate the cell level in a short time due to no-preprocessing of material.

Cell Cycle Analysis of Bovine Cultured Somatic Cells by Flow Cytometry

  • H.T. Cheong;D.J. Kwon;Park, J.Y.;J.W. Cho;Y.H. Yang;Park, T.M.;Park, C.K.;B.K. Yang;Kim, C.I.
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.69-69
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    • 2001
  • The cell cycle phase in which donor nuclei exist prior to nuclear transfer is an important factor governing developmental rates of reconstituted embryos. It was suggested that quiescent G0 and cycling G1 cells could support normal development of reconstituted embryos. In a quest of optimized donor nuclei treatment prior to nuclear transfer, this study was undertaken to examine the cell cycle characteristics of bovine fetal and adult somatic cells when cultured under a variety of culture treatments and the cell cycle change with the lapse of time after trypsinization. This was archived by measuring the DNA content of cells using flow cytometry, Cultured fetal fibroblast cells, adult skin and muscle cells, and cumulus cells were divided by 3 culture treatments; 1) grown to 60-70% confluency (cycling), 2) serum starved culture, 3) culture to confluency. Trypsinized cells were fixed by 70% ethanol and stained with propidium iodide. For one experiment, trypsinized cells were resuspended in DMEM+10% FBS and incubated for 1.5, 3 and 6 h with occasional shaking before ethanol fixation. Cell cycle phases were determined by flow cytometry enabling calculation of percentages of G0+G1, S and G2+M. The majority of cells were in G0+Gl stage regardless of origin of cells. Cultures that were serum starved or cultured to confluency contained significantly (P<0.05) higher percentages of cells in G0+G1 (89.5-95.4%). For every cell lines and culture treatments, percentages of cells in existing in G0+G1 increased with decreasing of the cell size from large to small. In the serum starved and confluency groups, about 98% of small cells were in G0+G1 Serum starved culture contained higher percentages of small-sized cells (38.5-66.9%) than cycling and confluent cultures regardless of cell lines (P<0.05). After trypsinization of fetal fibroblast and adult skin cells that were serum starved and cultured to confluency, the percentages of cells in G0+G1 significantly increased by incubation for 1.5(95.7-99.5%) and 3.0 h (95.9-98.6%). The results suggest that the efficient synchronization of bovine somatic cells in G0+G1 for nuclear transfer can be established by incubation for a limited time period after trypsinization of serum starved or confluent cells.

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Variation of Nuclear DNA Content in Interspecific Allium cepa L.×A. fistulosum L. hybrids and Their Successive Backcross Lines (양파와 파간의 종간잡종 F1과 여교잡계통의 핵 DNA 함량)

  • Kim, Cheol-Woo;Kim, Hwa-Young;Lee, Eul-Tai;Choi, In-Hu;Bang, Jin-Ki
    • Korean Journal of Breeding Science
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    • v.41 no.4
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    • pp.463-467
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    • 2009
  • Interspecific hybrid plants between Allium cepa L. (2n=2X=16) and A. fistulosum L. (2n=2X=16)and their backcross lines were developed by artificial pollination in order to introduce new desirable characters of A, cepa to A. fistulosum. The 2C nuclear DNA content has been estimated by flow cytometry in 5 Allium fistulosum inbreed lines, 2 interspecific hybrid lines of A. cepa${\times}$A. fistulosum and 34 their backcross lines $BC_1F_1$ to $BC_2F_2$, using propidium iodide (PI) as a fluorescence dye. Estimated 2C DNA values ranged from 22.2 pg to 23.7 pg in 5 A. fistulosum inbreed lines, 37.9 pg in F1 hybrid between A. cepa and A. fistulosum, 24.3 pg to 27.3 pg in 7 backcross lines in $BC_1F_1$, 21.9 pg to 24.4 pg in 9 $BC_1F_2$, 22.9 pg to 25.1 pg in 14 $BC_2F_1$, 22.6 pg to 23.4 pg in 4 $BC_2F_2$. This study showed mean 2C nuclear DNA content of $F_1$ hybrid was higher than their backcross progeny lines, while it was lower than female parental line, A. cepa (2C DNA=33.2 pg). Mean 2C DNA content of backcross lines, $BC_1F_1$ to $BC_2F_2$ was not significantly different but their 2C DNA contents in the more progress generation from $BC_1F_1$ to $BC_2F_2$ were reduced.

Characterization of Human Dental Pulp Cells from Supernumerary Teeth by Using Flow Cytometry Analysis (유세포 분석을 통한 과잉치 치수 유래 세포의 줄기세포 특성 연구)

  • You, Yonsook;Kim, Jongbin;Shin, Jisun;Lee, June-Haeng;Kim, Jongsoo
    • JOURNAL OF THE KOREAN ACADEMY OF PEDTATRIC DENTISTRY
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    • v.46 no.3
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    • pp.337-342
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    • 2019
  • The aim of this study was to analyze cells from human dental pulp tissue of impacted supernumerary teeth as stem cells with flow cytometry. Human dental pulp cells from 15 supernumerary teeth were identified their characteristics as stem cells by expression of mesenchymal stem cell markers through flow cytometry analysis at passage 3 and passage 10. Cluster of differentiation (CD) 73, CD 90, CD 34, CD 45 and STRO-1 cell surface markers were used to figure out characteristics of dental pulp stem cells from supernumerary teeth. At passage 3, the cell population showed positive expression of CD 73, CD90 and STRO-1, lacked expression of CD 34 and CD 45. At passage 10, CD 73, CD 90 and STRO-1 showed positive expression while CD 34 and CD 45 showed negative expression. This study indicated that dental pulp stem cells of supernumerary teeth had the properties of mesenchymal stem cells at both early and late passage. Impacted supernumerary teeth could be considered as a noble source of stem cells because of rapid growth and maintaining characteristics of stem cells until late passage.