• Title/Summary/Keyword: 형광액

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Comparative Studies of Digestive Fluid Protein of Silkworm Bombyx mori, Larvae reared on Mulberry Leaves and Artificial Diets (상엽육잠과 인공사료육잠의 소화액단백질의 비교연구 - 소화액 RFP를 중심으로 -)

  • 박희정;문재유
    • Journal of Sericultural and Entomological Science
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    • v.28 no.1
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    • pp.15-23
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    • 1986
  • This study has been carried out to acquire some basic informations on the digestive fluid of Silkworm, Bombyx mori for developing artficial diets. Silkworms reared on mulberry leaves and artificial diet, were used in this experiments. The results obtained are as follows ; 1. The Red Fluorescent protein was precipitated in 50% acetone solution and did not dissolved in n-butanol solution, but dissolved in methanol solution. 2. Electrophoretic analysis results of mulberry leaves rearing B. mori and artificial diet rearing B. mori, which has been treated with 50% acetone solution were as follows. i) There was distinct difference at the position of high mobility ii) Red Fluorescence was observed only at the position of first band of mulberry leaves rearing B. mori. iii) No different was shown in the electrophoretic patterns of mulberry leaves rearing B. mori on 5th instar 1st-3rd day, but some difference on 5th instar 4th, 5th day. 3. The RFP is the basic protein which has PI 8-9 according to the isoelectric electrophhoresis. 4. The SDS-polyacrylamide gel electrophoresis analysis showed that the molecular weight of RFP was 27,000. 5. The Sephadex G-75 chromatographic analysis showed that there was three peaks between number 16 and 28 in the mulberry leaves rearing B. mori chromatogram, but two peaks between number 18 and 31 in the artifical diet rearing B. mori chromatogram.

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Trial for Drug Susceptibility Testing of Mycobacterium tuberculosis with Live and Dead Cell Differentiation (세포 염색 방법을 이용한 결핵균 감수성 검사법)

  • Ryu, Sung-Weon;Kim, Hyun-Ho;Bang, Mun-Nam;Park, Young-Kil;Park, Sue-Nie;Shim, Young-Soo;Kang, Seongman;Bai, Gill-Han
    • Tuberculosis and Respiratory Diseases
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    • v.56 no.3
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    • pp.261-268
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    • 2004
  • Background : The resurgence of tuberculosis and outbreaks of multidrug resistant (MDR) tuberculosis have increased the emphasis for the development of new susceptibility testing of the Mycobacterium tuberculosis for the effective treatment and control of the disease. Conventional drug susceptibility testings, such as those using egg-based or agar-based media have some limits, such as the time required and difficulties in determining critical inhibitory concentrations, but these are still being used in many diagnostic laboratories because of no better lternatives, considering cost and accuracy. To overcome these limits, a rapid and simple method for new susceptibility testing, using live and dead assays, was applied for a bacterial cell viability assay to distinguish dead from live bacterial cells based on two-color fluorescence. Materials and Methods Strains : Forty strains were used in this study, 20 susceptible to all antituberculosis drugs and the other 20 resistant to the four first line antituberculosis drugs isoniazid, rifampicin, streptomycin and ethambutol. Antibiotics : The four antibiotics were dissolved in 7H9 broth to make the following solutions: $0.1{\mu}g\;isoniazid(INH)/m{\ell}$, $0.4{\mu}g\;rifampicin(RMP)/m{\ell}$, $4.0{\mu}g\;streptomycin(SM)/m{\ell}$ and $4.0{\mu}g\;ethambutol(EMB)/m{\ell}$. Results : Live and dead Mycobacterium tuberculosis cells fluoresced green and red with the acridin (Syto 9) and propidium treatments, respectively. These results are very well accorded with conventional drug susceptibility testing by proportional method on Lowensen-Jensen media (L-J) containing 4 drugs (INH, RMP, EMB and SM), showing a 93.7 % accordance rate in susceptible strains and 95% in resistant strains. Conclusion : The results of the drug susceptibility testing using the live and dead bacterial cell assay showed high accordance rates compared with the conventional proportion method on L-J. This finding suggests that the live and dead bacterial cell assay can be used as an alternative to conventional drug susceptibility testing for M. tuberculosis strains.

Rapid Detection of Fluorescent DNA on Denaturing Polyacrylamide Gel by Using Gel Scanner (겔스캐너를 이용한 변성아크릴아마이드 겔의 형광 DNA 검출)

  • Ku Ja-Hwan;Jeong Ji-Ung;Cho Young-Chan
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.50 no.spc1
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    • pp.228-230
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    • 2005
  • The denature polyacrylamide gel stain silver nitrate is used for routine nucleic acid detection. More sensitive stains, such as Vistra Green, SYBR Green are available to address a broad range of DNA applications requiring lower detection limits in polyacrylamide gel formats. Gel Scanners, laser-scanning instruments, provide sensitive fluorescence detection of DNA gel stains. We established one step fluorescent impregnation enhanced sensitivity with simple, rapid and low cost. We have applied this fluorescent staining procedure for the routine analysis of DNA profiles generated by SSR amplification.

Determination of Histamine by Reverse-Phase High Performance Liquid Chromatography with Fluorescence Detection (형광검출 역상 액체크로마토그래피에 의한 Histamine의 정량)

  • Ryu, Hee-Chun;Kim, Hyung-Ryong;Kim, Sang-Hyun;Kim, Dae-Ki;Lee, Young-Mi;Kim, Hyung-Min;An, Nyeon-Hyoung;Shin, Tae-Yong
    • Analytical Science and Technology
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    • v.9 no.2
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    • pp.198-202
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    • 1996
  • A rapid and simple method for the determination of histamine by reverse-phase high performance liquid chromatography with fluorescence detection was established. 9-Fluorenylmethyl chloroformate(FMOC) was used as fluorescent derivative reagent. The optimum conditions for the derivatization such as pH, reaction time and the concentration of FMOC were investigated. Linearity of calibration curve was obtained between $0.1{\mu}g/ml$ and $0.5{\mu}g/ml$(r=0.992) and the limit of detection was $0.01{\mu}g/ml$.

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Microanalysis of Vecuronium Bromide in Biological Fluids by Spectrofluorimetry and HPLC-Fluorescence Detection (형광광도법 및 HPLC-형광검출법에 의한 생체시료 중의 Vecuronium bromide의 미량분석)

  • Ko, Yong-Seok;Han, Sang-Soo;Shin, Tae-Yong;An, Nyeon-Hyoung;Ock, Chi-Wan
    • Analytical Science and Technology
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    • v.7 no.3
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    • pp.413-419
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    • 1994
  • The determination of the neuromuscular blocking agents vecuronium bromide(VeBr) in biological fluids has been investigated. The method depends on the formation of insoluble red complex between vecuronium bromide and rose bengal in aqueous layer. The amount of vecuronium bromide was calculated from that of extracted rose bengal which was determined by spectrofluorimetry or HPLC/fluorescence detection method. It was possible to analyze VeBr in the range of $2{\sim}32{\mu}g/ml$(r=0.998 for water soln., 0.999 for urine, 0.996 for plasma). This method was applied to the analysis of VeBr in biological fluids, urine and plasma.

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Changes in the Characteristics of Dissolved Organic Matter by Microbial Transformation and the Subsequent Effects on Copper Binding (생분해에 따른 용존 유기물질 성상 및 중금속 구리와의 결합특성 변화)

  • Jung, Ka-Young;Hur, Jin
    • Journal of Korean Society of Environmental Engineers
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    • v.34 no.1
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    • pp.49-54
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    • 2012
  • Microbial changes in the characteristics of dissolved organic matter (DOM) and the subsequent effects on the conditional stability constants of copper were investigated using 14 day-incubations of Pony Lake fulvic acid (PLFA), Suwannee River fulvic acid (SRFA) and the mixtures of the humic substances and glucose. After incubation, dissolved organic carbon (DOC) concentrations were diminished, and specific UV absorbance values and DOC-normalized fluorescence intensities increased. The microbial changes were minimal for the samples contaning humic substances only whereas they were much pronounced for the mixtures with glucose. The extent of the changes increased with a higher content of glucose in the mixtures. The same trend was observed even for glucose solution. Our results suggest that labile organic moieties may be transformed into more chromophoric and humidified components by biodegradation. For the mixture samples, the copper binding stability constants did not change or even decreased after incubation. Therefore, microbially induced enrichment of the fulvic- and humic-like carbon structures in DOM appears to result in little change or the decrease of the copper binding coefficients.

Fluorescence Assay for High Efficient Mass Screening of the Herbicides Inducing Rapid Membrane Peroxidation (막과산화를 신속히 유발하는 제초제의 고효율 대량스크리닝을 위한 형광검정법)

  • Kim, Jin-Seog;Kwon, Ok Kyung
    • Weed & Turfgrass Science
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    • v.4 no.4
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    • pp.308-314
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    • 2015
  • This study was conducted to establish a fluorescence assay system for high efficient mass screening of the herbicides causing rapid membrane peroxidation, based on the fact that peroxide in cellular leakage could be fluorometrically determined through the fuorescent compounds formed after reacting with homovanillic acid (HVA) and peroxidase (HRP). The assay procesure established in this study was as follows. Only single disc (4 mm diameter) excised from cucumber cotyledon is placed on the well containing test solution ($200{\mu}L$) with 96-well microplate. The plate is shaking-incubated for 8 h under light condition. Then after removing the cucumber disc, HVA and HRP are supplied in the medium buffer and incubated for 5 min at room temperature. Fluorescence values are determined at Ex 320 nm/Ex 425 nm. The higher fluorescence values are obtained in the treatment of chemical having higher herbicidal activity. Using this assay with 96-well microplates, a large number of herbicides inducing rapid membrane peroxidation seemed to be screened more efficiently than spectrophotometric microtiter assay reported previously.

Ion Exchange Separation and Spectrofluorometric Determination of Lanthanides in Nuclear Grade Material (이온 질환수지 및 형광분석법에 의한 핵급물질 중희토류원소의 분리정량)

  • Ki-Soo Cho;In-Suck Suh
    • Nuclear Engineering and Technology
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    • v.15 no.2
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    • pp.142-148
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    • 1983
  • Distribution coefficients between cation exchange resin (Dowex 50W$\times$8) and $\alpha$-hydroxyisobutric acid ($\alpha$-HIBA) are measured in order to separate traces of Sm, Eu, Gd and Dy from nuclear material. Individual separations are performed by pH gradient technique with 0.40M $\alpha$-HIBA from 3.40 to 3.60 in cation exchange resin after a group separation. Each of separated elements is determined with a fluorometric method except Gd by colorimetry. The results are applied to analyze Sm, Eu, Gd and Dy in magnesium diuranate (yellow cake).

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Effects of Nutrient Solution Strength on Growth and Nutrient Element Concentrations of Leaf Lettuce by Hydroponic Culture under Artificial Light (인공광을 이용한 수경재배에서 배양액 농도가 상추의 생장과 배양액 양분 농도에 미치는 영향)

  • Kim, D.E.;Lee, W.Y.;Heo, J.W.;Lee, G.I.;Kang, D.H.;Woo, Y.H.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.19 no.1
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    • pp.5-14
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    • 2017
  • This study was conducted to investigate the effects of nutrient solution strength on growth and nutrient element concentrations in leaf lettuce (Lactuca sativa L. cv. 'Dduksum') by hydroponic culture under fluorescent lamp and LED. Leaf lettuce were grown in closed hydroponic cultivation systems supplied with 1/2, 1 and 2 strength of nutrient solution recommended by horticultural experiment station in Japan. The growth of 'Dduksum' was highest in the 2 strength of standard nutrient solution. The amount of nutrient element in the recycled nutrient solution was higher at 2, 1 and 1/2 strength of nutrient solution. The concentration of NO3-N, Ca2+, Mg2+ in the recycled nutrient solutions increased in 1 and 2 strength of nutrient solution but that of NH4-N decreased gradually in 1/2 and 1 strength of nutrient solution. The concentration of K, Ca, Mg in leaf lettuce was maintained in the normal range, whereas the concentration of phosphorous was 1.3 to 1.6%, which was higher than proper range. As the concentration of NH4-N decreases gradually in all the treatments, it is necessary to raise the rate of NH4-N or add it.

Development of Techniques for Testicular Germ Cell Transplantation in Pigs (돼지에 있어서 정소 생식세포의 이식 기법 개발)

  • Kim, Byung-Gak;Lee, Yong-An;Kim, Bang-Jin;Kim, Ki-Jung;Min, Kwan-Sik;Lee, Jang-Hee;Ryu, Jae-Weon;Kim, In-Cheul;Ryu, Buom-Yong
    • Reproductive and Developmental Biology
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    • v.32 no.3
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    • pp.193-198
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    • 2008
  • The current study was designed to extend the technique of spermatogonial transplantation to economically important pig model We evaluated the efficiency of pig to pig transplantation. Donor testis cells were harvested from testes obtained at castration of 10- to 14-day-old boars and were labeled with fluorescent marker(PKH26) before transplantation. The presence of infused dye or labeled pig testicular cells was confirmed in the seminiferous tubules from recipient pig. The most effective procedure of intratubular germ cell transfer was to insert an fine needle ($21{\sim}25$ gauge) through the cauda epididymis and testis into the rete testis under ultrasound guidance. Infusion of $5{\sim}7ml$ of dye solution or cell suspension could fill the rete and up to 50% of seminiferous tubules of 14-week-old boars. Testis were examined for the presence and localization of labeled donor cells immediately after transplantation and labeled donor cells were found in numerous seminiferous tubules from recipient pig testes. These results indicate that germ cell transplantation is feasible in recipient pig testis. This study represents successful spermatogonial transplantation between individual animals in a livestock species.