• Title/Summary/Keyword: 96-well plate

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Antifungal activities of sulphamide and dicarboximide fungicides against Botrytis cinerea in several in vitro bioassays (여러 종류의 in vitro 생물검정에서 Botrytis cinerea에 대한 sulphamide계와 dicarboximide계 살균제의 활성 특성)

  • Choi, Gyung-Ja;Kim, Heung-Tae;Kim, Jin-Cheol;Cho, Kwang-Yun
    • The Korean Journal of Pesticide Science
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    • v.3 no.3
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    • pp.37-44
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    • 1999
  • Two sulphamide (dichlofluanid and tolylfluanid) and three dicarboximide fungicides (iprodione, vinclozolin, procymidone) were used to investigate the correlation between in vitro antifungal activities and in vivo disease controlling activities against Botrytis cinerea, a causal agent of tomato gray mold and to develop efficient in vitro assays. They controlled effectively the development of tomato gray mold disease in vivo and their controlling activities were similar one another. However, several in vitro assays revealed that their in vitro antifungal activities were quite different between sulphamide and dicarboximide fungicides; the formers showed stronger inhibition activities for spore germination than the latters, whereas the formers inhibited mycelial growth less severely than the latters. The results indicate that the fungicides having different modes of action can show different in vitro antifungal activities according to in vitro assays, even if they have similar in vivo disease controlling activities. On the other hand, two rapid and efficient in vitro assays named Microtiter plate methods I (MPM I) and II (MPM II) were developed for the evaluation of fungicides for inhibitory activities against spore germination and mycelial growth of B. cinerea, respectively. The antifungal activities of five fungicides of two chemical groups in MPM I and II were correlated with the inhibitory activities against spore germination and mycelial growth using solid media, respectively.

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High Throughput Screening for Searching a New Inhibitors of Acetolactate Synthase (Acetolactate synthase에 대한 고효율 활성 측정방법 및 신규 저해제 탐색)

  • Park, S.H.;Lee, K.H.;Choi, J.S.;Pyon, J.Y.;Cho, K.Y.;Hwang, I.T.
    • The Korean Journal of Pesticide Science
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    • v.5 no.3
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    • pp.41-46
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    • 2001
  • This study was conducted to develop a high throughput system for screening acetolactate synthase(ALS) inhibitors, and to detect basic mother molecules for developing new novel herbicide candidates. The high throughput screening (HTS) method using 96-well plate and microplate reader was developed. This method is 8 times more effective than basic technique in one cycle per person. Futhermore, considering for less than 1/10 volume of materials required for ALS test and enzyme kinetics with 16 times faster speed compared to those of former procedure, this HTS method has more than 100 times higher efficacy than basic system in a consecutive procedure. We discovered 11 new ALS inhibitors such as 2-oxoglutaric acid, aminooxyacetic acid, azelaic acid, citric acid, cyanuric fluoride, itaconic acid, malonic acid, niclosamide, oxalic acid, glyoxylic acid, and suramin from 107 commercial plant-specific inhibitors using this technique. We hope these results might be useful to discover lead compounds for developing new novel herbicide candidate.

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Spectrophotometric Assay for Determination of Chlorogenic Acid Using Green Pigment Formation and Quantitative Analysis of Chlorogenic Acid in Blueberry Leaf (녹색반응을 이용한 클로로겐산의 함량측정을 위한 흡광도 분석법과 블루베리 잎에 함유된 클로로 겐산의 함량분석)

  • Chung, Dong-Min;Chung, Young-Chul;Chun, Hyo-Kon
    • Journal of Life Science
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    • v.21 no.4
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    • pp.610-612
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    • 2011
  • We developed a spectrophotometric assay for the quantitative determination of chlorogenic acid based on the formation of green pigment at $50^{\circ}C$ under glycine and alkaline conditions in 96-well plates. The formation of green pigment was linear with a series of chlorogenic acid concentration (0-$300\;{\mu}M$). Using this method, the content of chlorogenic acid (12.42 mg/g dry weight) in the leaves of blueberry was quantified. This method is high-throughput, cost-effective, rapid, and easy to perform.

Development of a Zero Discharge and Reuse System for Rural Areas (농촌지역을 위한 무방류 재이용시스템 개발)

  • Hong, Min;Choi, Kyung-Sook
    • Journal of The Korean Society of Agricultural Engineers
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    • v.51 no.6
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    • pp.91-96
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    • 2009
  • This study describes a zero discharge and reuse system developed for rural areas. The purpose of the system is decontamination of used irrigation water for down-stream usage and reuse of wastewater in rural villages for preventing water shortage problem expected to happen in near future. The system consists of anoxic, FES (Ferrous Electricity System), Oxic, Cralifier processes. The main feature of the system is to remove phosphorous by using Fe-ionizing module. Indoor experiments were undertaken with a trial product of the system to test its performance. The removal capacities of T-P, T-N, and BOD were examined. Also the proper time for the replacement of iron plate module was tested as well as the efficiency of T-P removal rate based on the usage of an automatic washing system for the iron plate. As results, the system showed very good water purification performances through obtaining the results of over 90% removal rates from T-P, BOD, and 67% from T-N. The proper time period for replacement of iron plate was maximum 2 years, and also efficiency of T-P removal rate found to be greatly influenced by the usage of an automatic washing system from the test.

Study on the Production and Management of Aquatic Animal : Application of ELISPOT-Assay for the Detection of Antibody Secreting Cells in Flounder, Paralichthys olivaceus (수산생물의 생산과 관리에 관한 기초연구 : ELISPOT 기법을 이용한 넙치의 항체생성 세포분석)

  • HA Jai Yi;PARK Jun-Hyo;KIM Myoung Sug;CHUNG Joon-Ki;JEONG Hyun Do
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.32 no.4
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    • pp.420-426
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    • 1999
  • We examined the immune response in flounder, Paralichthys olivaceus, with immunization of formalin killed Edwardsiella tarda as an antigen. The ELISPOT-assay (enzyme-linked immunospot assay) was optimized technically and applied to count the number of total and specific antibody secreting cells (TASC and SASC) in lymphocytes of different lymphatic organs. Incubation of lymphocytes on 96 well plate for more than 2.5hrs came out enough time in ELISPOT-assay for counting the antibody secreting cells in the anterior kidney and spleen. However, too much of plate-coated antigen or rabbit anti-flounder immunoglobulin for SASC or TASC counting, respectively, was appeared to decrease the sensitivity of the assay system. Specificity of the system was also confirmed by the absence of TASC in lymphocytes treated with cycloheximide to prevent protein synthesis. The peak numbers of SASC appeared at wk 3 post immunization after that there was a sharp decrease and reached to almost zero at wk 7. In the spleen and kidney, the timing and numbers of SASC on peak response were concurrent without preferential organ distribution. The specific antibody level in the sera increased rapidly between wk 2 and 3 after immunization, i.e. like the specific cellular response found with ELISPOT-assay on that period, However, the remained high level of specific serum antibody from wk 5 after immunization until the end of experiment was clearly distinguishable from the kinetics of SASC response decreased sharply.

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Optimized study of an in vitro 3D culture of preantral follicles in mice

  • Hehe Ren;Yingxin Zhang;Yanping Zhang;Yikai Qiu;Qing Chang;Xiaoli Yu;Xiuying Pei
    • Journal of Veterinary Science
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    • v.24 no.1
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    • pp.4.1-4.16
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    • 2023
  • Background: In vitro culture of preantral follicles is a promising technology for fertility preservation. Objectives: This study aims to investigate an optimized three-dimensional (3D) fetal bovine serum (FBS)-free preantral follicle culture system having a simple and easy operation. Methods: The isolated follicles from mouse ovaries were randomly divided in an ultra-low attachment 96-well plates supplement with FBS or bovine serum albumin (BSA) culture or encapsulated with an alginate supplement with FBS or BSA culture. Meanwhile, estradiol (E2) concentration was assessed through enzyme-linked immunosorbent assay of culture supernatants. The diameter of follicular growth was measured, and the lumen of the follicle was photographed. Spindle microtubules of oocytes were detected via immunofluorescence. The ability of oocytes to fertilize was assessed using in vitro fertilization. Results: The diameters were larger for the growing secondary follicles cultured in ultra-low attachment 96-well plates than in the alginate gel on days 6, 8, and 10 (p < 0.05). Meanwhile, the E2 concentration in the BSA-supplemented medium was significantly higher in the alginate gel than in the other three groups on days 6 and 8 (p < 0.05), and the oocytes in the FBS-free system could complete meiosis and fertilization in vitro. Conclusions: The present study furnishes insights into the mature oocytes obtained from the 3D culture of the preantral follicle by using ultra-low attachment 96-well plate with an FBS-free system in vitro and supports the clinical practices to achieve competent, mature oocytes for in vitro fertilization.

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.

Development of Gridding Robot System for Genome Research (유전체 연구용 그리딩 로봇 시스템의 개발)

  • 추창환;서동현;김찬수;박지영;임용표;김기대
    • Journal of Biosystems Engineering
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    • v.26 no.4
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    • pp.391-398
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    • 2001
  • A robot system for clone replication and gridding, which is a preliminary state of the genome research, was developed and evaluated its performance. This gridding robot system consisted of 1) a gridding heat that replicated the clone, 2) a manipulator, as a part of body of robot, which transferred the gridding head along x-, y-, z-axis, 3) a well plate arranging board, 4) a sterilization unit, and 5) a control unit. Performance of the system was evaluated with 1) repeatability of the robot system, 2) clone replication efficiency, 3) time requirement of the replication, and 4) sterilization efficiency. The repeatability error of the robot system showed 0.219 mm and 0.094 mm in the direction of x- and y-axis, respectively. The success rate of the clone replication with the gridding head was 100% on the membrane filter. The time required for the replication was four minutes and fifty-five seconds from the four 96 well plates to a 384 well plate meanwhile the required time with well experienced hand labor was three minutes thirty-five seconds. The gridding operation of clone could not be done by hand labor and the required time with robot system for the gridding on the membrance filter with the control program 5$\times$5: 1 copy and 384 gridding pins was twenty minutes and twenty-five seconds. The efficiency of the sterilization was considered to be satisfactory since no growth of fungi was found around the area of replication in the membrane filter.

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Modified CTAB DNA Methods for efficient DNA extraction from Rice (Oryza sativa L.) (벼 분자육종을 위한 CTAB DNA 추출 시스템 개량)

  • Lee, Jong-Hee;Kwak, Do-Yeon;Yeo, Un-Sang;Kim, Choon-Song;Jeon, Myeong-Gi;Kang, Jong-Rae;Park, Dong-Soo;Shin, Mun-Sik;Oh, Byeong-Geun;Hwang, Hung-goo
    • Korean Journal of Breeding Science
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    • v.40 no.3
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    • pp.286-290
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    • 2008
  • Many important traits have been tagged allowing plant breeders to apply marker assisted selection (MAS) in rice. PCR itself is simple to set up, and requires little hands-on time. However, a crucial limiting step of MAS programs is the reliable and efficient extraction of DNA which can be performed on thousands of individuals. In this study, We describe a modification of the DNA extraction method, in which cetyltrimethylammonium bromide (CTAB) is used to extract DNA from leaf tissues for suitable MAS in rice. We followed the standard 2% CTAB extraction method in all the procedure. In addition we used the 1.2 ml 8-strip tube instead of 1.5 ml E-tubes to fit the 8-multichannel pipette and employ the 96 well plate to use the swing bucket centrifuge. Our modified CTAB DNA extraction method offers several advantages with respect to traditional and simple methods. 1) adult leaf samples collected in paddy field are applicable. 2) 96 leaf samples can be homogenized only one-time by using tungsten carbonate bead and 96well block. 3) semiautomatic loading method using 8-multichannel pipette from DNA extraction to electrophoresis of PCR products. 4) our system can extract about 400 leaf samples per day by only one technicion. Therefore, this method could be useful for marker assisted breeding in rice.

Determination of an Optimum Orbiting Radius for an Oil-Less Scroll Air Compressor

  • Kim, Hyun-Jin;Lee, Yong-Ho;Kwon, Tae-Hun
    • International Journal of Air-Conditioning and Refrigeration
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    • v.16 no.4
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    • pp.124-129
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    • 2008
  • Design practice has been made on an oil-less scroll air compressor as an air supply device for a 2 kW fuel cell system where air pressure of 2 bar and flow rate of 120 liter/min are required. Basic structure of the scroll compressor includes double-sided scroll wrap for the orbiting scroll driven by two crankshafts connected to each other by a timing belt. These features can eliminate thrust surface which otherwise would produce frictional heat and jeopardize reliable operation of the orbiting scroll and the scroll element's deformation as well. This study focuses on optimum scroll wrap design; orbiting radius has been chosen as an independent design parameter. As the orbiting radius changes, scroll sizes such as scroll base plate and discharge port diameters change accordingly. Gas compression-related losses and mechanical loss also change with the orbiting radius. With a scroll base plate diameter of 120mm at most and discharge port of at least 10mm, the orbiting radius should be within the range of 2.5-4.0mm. With this range of the orbiting radius, it was estimated by performance analysis that the compressor efficiency reached to a maximum of ${\eta}_c$=96% at the orbiting radius of $r_s$=3.5mm for the scroll wrap height-to-thickness ratio of h/t=5.