• 제목/요약/키워드: DAPI

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Application of DNA Content and Total Protein Concentration to Predict Blooms Caused by Cochlodinium polykrikoides (Dinophyceae) in Korean Coastal Waters (total DNA 및 단백질 함량변화에 의한 C. polykrikoides 조기적조 예측 응용)

  • Cho, Eun-Seob;Park, Yong-Kyu
    • Journal of Life Science
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    • v.14 no.2
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    • pp.255-262
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    • 2004
  • We applied nuclear DNA content stained with 4'-6'-diamidino-2-phenylindole (DAPI) and total protein concentration to predict the existence of Cochlodinium polykrikoides before huge blooms occurred, based on a short-term survey at sites in the South Sea. Fluctuations in environmental conditions and nutrients (nitrate, nitrite, and phosphate) were of a similar range, regardless of sampling sites or early and middle field observations. However, C. polykrikoides abundance was significantly different depending on the station, with a higher cell density of 34, 62, and 57 cells L$^{-1}$ at Stn C2, C5, and C6, respectively than what was found in early August, 2000. In mid August, 2000, the highest cell density of 547 cells L$^{-1}$ at Stn C3 was observed. The relationship between C. polykrikoides abundance, DAPI-stained DNA content, and total protein concentration was a positive correlation coefficient, in particular a higher positive correlation was exposed to even a smaller abundance of C. polykrikoides. These results suggest that DNA stained by DAPI and total protein concentration could play an important index in easily predicting the presence of C. polykrikoides before blooms.

Application of Image Analysis System for Red Tide Organisms

  • Cho Eun Seob;Kang Yoon Mi;Kim Gwang Hoon
    • Fisheries and Aquatic Sciences
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    • v.2 no.2
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    • pp.172-175
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    • 1999
  • Relative DNA contents in some harmful algae were measured using DAPI staining and image analysis system. This method was useful to identify some morphologically similar species and isolates from harmful algal blooms (HABs). In exponential phase, Prorocentrum micans had higher relative DNA content (RD) of $1.83\pm0.52$ than any other isolates, followed by Cochlodinium polykrikoides $(1.10\pm0.46)$ Alexandrium tamarense $(0.93\pm0.32)$ Gyrodinium impudicum $(0.56\pm0.17)$, Scrippsiella trochoidea $(0.41\pm0.26)$ and P. minimum$(0.05\pm0.01)$. When they were fixed with Lugol's solution, it was difficult to d,iscern C. polykrikoides from G. impudicum under the light microscope, but the DNA contents were quite different in two species. C. polykrikoides contained about twice as much RD as G. impudicum under the same culture conditions and exponential phase. DAPI­stained DNA feature in C. polykrikodes showed concentrated in the peripheral part of the cell, but in G. impudicum showed a compact structure in the central part. Although A. tamarense and S. trochoidea were morphologically similar under the light microscope, nuclear DNA content of A. tamarense was twice as much as that of S. trochoidea.

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Distribution of Mycoplasma in Witches'-broom Infected Jujube Tissue (빗자루병(病)에 감염(感染)된 대추나무 조직내(組織內)의 마이코플라스마 분포(分布))

  • La, Yong Joon;Lee, Deok Jae
    • Journal of Korean Society of Forest Science
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    • v.67 no.1
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    • pp.28-30
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    • 1984
  • Distribution of mycoplasma-like organisms (MLO) in the phloem tissue of witches'-broom infected jujube trees was investigated by fluorescence microscopy applying new fluorochrome DAPI (4'-6-diamidino-2-phenylindole, 2HCl). MLO were detected from the phloem of leaf and stem sections of diseased branches exhibiting typical witches'-broom symptom but not detected from those of symptomless branches of an infected tree. MLO were also present in the healthy looking leaves frequently found in the diseased shoots. Fluorescence microscopy revealed the presence of MLO in the phloem of root sections of infected trees.

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Life Cycle of Heterotrophic Dinoflagellate Cryptoperidiniopsis brodyi (Dinophyceae) (Cryptoperidiniopsis brodyi (Dinophyceae)의 생활사)

  • Park, Tae-Gyu;Park, Young-Tae;Bae, Heon-Meen
    • Journal of Environmental Science International
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    • v.18 no.1
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    • pp.9-14
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    • 2009
  • Pfiesteriaand Pfiesteria-like organisms were reported to be linked to major fish kills(involving well over a billion fish) in North Carolina and Maryland estuaries on the U.S. east coast during the 1990s. Occurrences of these species have been recently reported from Korean waters including Chinhae Bay and the coast of Yeosu. In this study, the life cycle of Cryptoperidiniopsis brodyi and Pfiesteria piscicida were examined using DAPI staining. Their excystment and growth were stimulated directly by the addition of prey cells such as Rhodiminas salina. Amoeboid stages in C. brodyi and P. piscicida were never observed in culture, even after addition of filter-sterile fish mucus and tissue. The dominant life cycle stages consisted of motile flagellated zoospores and cysts. A typical dinoflagellate life cycle was demonstrated by direct observation and DAPI staining.

Comparison of Conditions for Cell Death-Inducing Agents Using a High Throughput-Compatible Nuclear Staining Assay (핵 염색을 이용한 세포사멸 유도물질 스크리닝의 조건 비교)

  • Lee, Sang-Han
    • Journal of Life Science
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    • v.18 no.9
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    • pp.1312-1315
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    • 2008
  • High throughput-drug screening plays a pivotal role for early stage of drug discovery process. In the course of assay development for screening of cell death-inducing agents, a protocol that is simple, time-saving, and high throughput-compatible was designed which was confirmed that the protocol can be worked by a HTS-compatible machine. In 96-well format, PC-3 cancer cells (1${\times}10^{4}$ cells/ml) were cultured for 24 hr. After 24 h-incubation with various medicinal plants extracts, the cells were then stained with DAPI for 30 min. The fluorescence intensity of the stained cells was measured semi-automatically with a multilabel counter. To test whether the present assay system effectively works, we screened about 850 medicinal plant extracts, and selected 1 positive crude extracts that contained cell death-inducing activity. These results suggest that the protocol is highly amenable to HTS implementation for a cell death-inducing agent(s).

Protective effect of platelet-rich plasma against cold ischemia-induced apoptosis of canine adipose-derived mesenchymal stem cells

  • Suji Shin;Sung-Eon Kim;Seong-Won An;Seong-Mok Jeong;Young-Sam Kwon
    • Korean Journal of Veterinary Research
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    • v.64 no.1
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    • pp.2.1-2.8
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    • 2024
  • This study was performed to assess the antiapoptotic effect of canine platelet-rich plasma (PRP) treated on the canine adipose-derived mesenchymal stem cells (cMSCs) under cold ischemic conditions. The effect of preventing apoptosis of cMSCs was evaluated in the apoptotic condition induced by cold ischemic injury in vitro. To determine the progression of apoptosis, the changes in cell nucleus were observed using 4',6-diamidino-2-phenylindole (DAPI) fluorescence staining. In addition, we examined the mitochondrial membrane potential (MMP) and caspase-3 activity. When the cold hypoxic injury was applied to cMSCs, the apoptotic change was observed by DAPI staining, mitochondrial staining for MMP, and caspase-3 assay. PRP significantly decreased the number of apoptotic cells. Nuclear shrinkage and fragmentation of apoptotic cells in control groups were observed by DAPI staining. The MMP was recovered by the treatment of PRP. In addition, when the luminescence intensity was measured for caspase-3 activity, the value was significantly higher in the PRP treated groups than the control groups. The results of this study showed that the PRP may have a beneficial effect on apoptosis induced by cold ischemic injury.

The Morphology Study of Organ Surface BongHan Ducts and Corpuscle (장기표면의 내외봉한관과 봉한소체의 형태학적 관찰)

  • Ahn, Seong-Hun;Kim, Min-Su;Lee, Sang-Hun;Kwon, O-Sang;Kim, Jae-Hyo;Soh, Kwang-Sup;Sohn, In-Chul
    • Korean Journal of Acupuncture
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    • v.26 no.1
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    • pp.79-84
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    • 2009
  • Objective : In 1960's Bonghan Kim's team found BongHan(BH) ducts which were presumed as acupuncture meridians and BH corpuscles. They asserted Bonghan theory and SanAl theory which was involved in cell division and cell restoration. However, many other experiments which had been operated to demonstrate and find the existence of BH ducts had failed because of the secret of blue stain drugs. During the last several years, BongHan theory has been revived through experimental researches to find the anatomical structures of BH ducts and corpuscles by Soh's Biomedical Physics Lab. Soh's research team used the staining with Janus Green B, Alcian blue, nanoparticles and Acridine Orange. We used DAPI staining to find the existence of BH ducts and the corpuscles and to observe nuclear arrangement. Methods : We used japan white rabbits as experimental animals. BH ducts and corpuscles were stained with DAPI. The nucleus configuration in BH ducts stained with DAPI were observed with microscope. Results : In this study, we found thread like structures in silver white color distinguished from the blood vessels, nerves and lymph vessels. These thread like vessels in the linear duct shape were connected to same colored mass in the ball shape. Thread like structures we found could be separated easily from the surrounding other organ mass. The nuclei of the thread like structure in DAPI staining, are about 10${\sim}$20${\mu}m$ length, in rod shape and linear arrangement. Conclusion : We concluded that the thread like structure we found was same vessel reported by Soh's research team, BongHan ducts and corpuscle.

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Characterization of rDNAs and Tandem Repeats in the Heterochromatin of Brassica rapa

  • Lim, Ki-Byung;de Jong, Hans;Yang, Tae-Jin;Park, Jee-Young;Kwon, Soo-Jin;Kim, Jung Sun;Lim, Myung-Ho;Kim, Jin A;Jin, Mina;Jin, Yong-Moon;Kim, Seog Hyung;Lim, Yong Pyo;Bang, Jae-Wook;Kim, Ho-Il;Park, Beom-Seok
    • Molecules and Cells
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    • v.19 no.3
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    • pp.436-444
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    • 2005
  • We describe the morphology and molecular organization of heterochromatin domains in the interphase nuclei, and mitotic and meiotic chromosomes, of Brassica rapa, using DAPI staining and fluorescence in situ hybridization (FISH) of rDNA and pericentromere tandem repeats. We have developed a simple method to distinguish the centromeric regions of mitotic metaphase chromosomes by prolonged irradiation with UV light at the DAPI excitation wavelength. Application of this bleached DAPI band (BDB) karyotyping method to the 45S and 5S rDNAs and 176 bp centromere satellite repeats distinguished the 10 B. rapa chromosomes. We further characterized the centromeric repeat sequences in BAC end sequences. These fell into two classes, CentBr1 and CentBr2, occupying the centromeres of eight and two chromosomes, respectively. The centromere satellites encompassed about 30% of the total chromosomes, particularly in the core centromere blocks of all the chromosomes. Interestingly, centromere length was inversely correlated with chromosome length. The morphology and molecular organization of heterochromatin domains in interphase nuclei, and in mitotic and meiotic chromosomes, were further characterized by DAPI staining and FISH of rDNA and CentBr. The DAPI fluorescence of interphase nuclei revealed ten to twenty conspicuous chromocenters, each composed of the heterochromatin of up to four chromosomes and/or nucleolar organizing regions.

Heterosigma akasiwo의 핵형분석을 통한 생활사 연구를 위한 DAPI이용 기법

  • Lee Ju Yeon;Han Myeong Su
    • Proceedings of the Korea Society of Environmental Biology Conference
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    • 2003.11a
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    • pp.121-124
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    • 2003
  • The goals of this study is to elucidate life cycle and to detect genetic differences within a single species of Heterosigma akashiwo. To elucidate life cycle of H. akashiwo, have to study of benthic stage and vegetative cell. So we studied identification of H. akashiwo cyst. The relative contents of DNA in nuclei were determined in Heterosigma akashiwo. Different stages of the life history were obtained from culture and natural sediments, and examined by microfluorometry after staining with the DNA-specific fluorochrome 4'-6-dianudubi-2-phenylindole(DAPI). Large cells mainly in exponensial stage, while small cell, pre-encystment cells(\ulcorner\ulcorner), showed in the end of the late growth stage. Type of DNA content showed the different with growth stage. Usually the small cell has the high level of IOD.

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The Changes of Bacillus spp. in Municipal Wastewater Treatment Plant with B3 process. (B3 공법을 사용하는 하수종말처리장에서 Bacillus 속 세균의 변화)

  • 안태석;홍선희;김옥선;유재준;전선옥;최승익
    • Korean Journal of Microbiology
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    • v.37 no.3
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    • pp.209-213
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    • 2001
  • By FISH and spore staining methods, the changes of Bacillus spp. numbers were investigated at municipal waste water treatment system, which operated with B3 process. Total bacterial numbers, which is the sum ofDAPI bound cells and numbers of spore, were ranged TEX>$3.2{\times}10^8 ~ 4.4{\times}10^8 $ cells/ml. The Bacilli numbers whichis the sum of probe S-G-Bacill-0597-a-A-22 bound cells and numbers of spore were from $0.3{\times}10^7 ~ 8.4{\times}10^7 $ cells/ml. The ratios of Bacillus sp. to total bacteria were ranged from 8 to 60%. Among 4 aeration tanks, thefirst tank had shown the highest proportions of Bacillus sp. And third and fourth tanks, the numbers of sporewere increasing.

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