• Title/Summary/Keyword: microplate method

검색결과 59건 처리시간 0.02초

$_L$ =lysine에 의한 Microcystis sp.의 선택적 성장억제 (The Effect of $_L$=lysine on Growth Inhibition of Microcystis sp.)

  • 송석환;신규철;한명수;최영길
    • 환경생물
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    • 제21권2호
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    • pp.216-221
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    • 2003
  • 본 연구는 $_L-lysine \; (_{-2}$, 6-Diaminohexanoic acid)의 영향을 받는 남조류와 남조류에 영향을 주는 $_L-lysine $의 농도를 파악 하고자 20종의 Microcystis.를 실험에 사용하였다. 남조류의 생장 억제 능력은 double-layered agar method와 microplate method를 이용하여 측정하였다. L-lysine의 농도를 100-${\mu}g\; ml-1~300 ${\mu}g\; ml^{-1}$이상에서 처리한 경우 Microcystis ichthyoblabe NIER-10021외 7종의 Microcystis sp.에서 투명대가 형성되었다. Microplate method서는10~500${\mu}g\; ml^{-1}$의 농도에서 생장억제 및 lysis를 나타내어TEk. 그리고 Microcystis viridis NIER-10020 외 3종은 10${\mu}g\; ml^{-1}$이하의 낮은 농도에서도 생장억제 및 분해를 나타내는 높은 활성을 보였다.

Microplate Reader를 이용하여 측정한 혈액의 엽산 농도와 실측량 기록법에 의한 엽산 섭취량 (Blood Folate Level Determined by a Microplate Reader and Folate Intake Measured by a Weighted Food Record)

  • 현태선;한영희;임은영
    • 대한지역사회영양학회지
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    • 제4권4호
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    • pp.512-520
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    • 1999
  • Microbiological method using a 96-well microplate reader for folate assay was established, and folate intake and blood folate concentrations of 23 female college students were assessed. To evaluate folate intake, dietary data were collected by a 3-day weight food record, and serum and RBC folate concentrations were measured by the new method. The coefficient of variation for the new method was less than 10%. Mean daily folate intake of the subjects was 126.7${\mu}g$ which is only 50.7% of the RDA. The mean concentrations of serum and RBC folate were 7.46ng/ml and 294.4ng/ml, respectively, which were within the normal range. These results indicate that folate intake seems to be underestimated due to incomplete food composition database. Therefore, folate database should be appropriately in order to asses folate intake accurately.

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Microplate Assay Measurement of Cytochrome P450-Carbon Monoxide Complexes

  • Choi, Suk-Jung;Kim, Mi-Ra;Kim, Sung-Il;Jeon, Joong-Kyun
    • BMB Reports
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    • 제36권3호
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    • pp.332-335
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    • 2003
  • Cytochrome P450 in microsomes can be quantitated using the characteristic 450 nm absorption peak of the CO adduct of reduced cytochrome P450. We developed a simple microplate assay method that is superior to previous methods. Our method is less laborious, suitable for analyzing many samples, and less sensitive to sample aggregation. Microsome samples in microplate wells were incubated in a CO chamber rather than bubbled with CO gas, and then reduced with sodium hydrosulfite solution. This modification allowed a reliable and reproducible assay by effectively eliminating variations between estimations.

A Colorimetric Microplate Assay Method for High Throughput Analysis of Lipase Activity

  • Choi, Suk-Jung;Hwang, Jung-Min;Kim, Sung-Il
    • BMB Reports
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    • 제36권4호
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    • pp.417-420
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    • 2003
  • The present work describes a colorimetric microplate assay for lipase activity based on the reaction between 5,5'-dithiobis(2-nitro benzoic acid) (DTNB) and the hydrolysis product of 2,3-dimercapto-1-propanol tributyrate (DMPTB). Reaction mixtures containing DTNB, DMPTB, and lipase were prepared in microplate wells, and the absorbance at 405nm was recorded after incubation at $37^{\circ}C$ for 30 min. A linear relationship was obtained in the range of 0.1-1 U of lipase activity by this method. The reaction conditions were also optimized for the range of 0.01-0.1 U or 1-10 U. When assaying crude tissue extracts, the reaction of DTNB with non-specific reducing agents created a major source of error. However, this error was corrected by the use of blank samples that did not contain DMPTB.

A Rapid Method for Estimating the Levels of Urinary Thiobarbituric Acid Reactive Substances for Environmental Epidemiologic Survey

  • Kil, Han-Na;Eom, Sang-Yong;Park, Jung-Duck;Kawamoto, Toshihiro;Kim, Yong-Dae;Kim, Heon
    • Toxicological Research
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    • 제30권1호
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    • pp.7-11
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    • 2014
  • Malondialdehyde (MDA), used as an oxidative stress marker, is commonly assayed by measuring the thiobarbituric acid reactive substances (TBARS) using HPLC, as an indicator of the MDA concentration. Since the HPLC method, though highly specific, is time-consuming and expensive, usually it is not suitable for the rapid test in large-scale environmental epidemiologic surveys. The purpose of this study is to develop a simple and rapid method for estimating TBARS levels by using a multiple regression equation that includes TBARS levels measured with a microplate reader as an independent variable. Twelve hour urine samples were obtained from 715 subjects. The concentration of TBARS was measured at three different wavelengths (fluorescence: ${\lambda}-_{ex}$ 530 nm and ${\lambda}-_{em}$ 550 nm; ${\lambda}-_{ex}$ 515 nm and ${\lambda}-_{em}$ 553 nm; and absorbance: 532 nm) using microplate reader as well as HPLC. 500 samples were used to develop a regression equation, and the remaining 215 samples were used to evaluate the validity of the regression analysis. The induced multiple regression equation is as follows: TBARS level (${\mu}M$) = -0.282 + 1.830 ${\times}$ (TBARS level measured with a microplate reader at the fluorescence wavelengths ${\lambda}-_{ex}$ 530 nm and ${\lambda}-_{em}$ 550 nm, ${\mu}M$) -0.685 ${\times}$ (TBARS level measured with a microplate reader at the fluorescence wavelengths ${\lambda}-_{ex}$ 515 nm and ${\lambda}-_{em}$ 553 nm, ${\mu}M$) + 0.035 ${\times}$ (TBARS level measured with a microplate reader at the absorbance wavelength 532 nm, ${\mu}M$). The estimated TBARS levels showed a better correlation with, and are closer to, the corresponding TBARS levels measured by HPLC compared to the values obtained by the microplate method. The TBARS estimation method reported here is simple and rapid, and that is generally in concordance with HPLC measurements. This method might be a useful tool for monitoring of urinary TBARS level in environmental epidemiologic surveys with large sample sizes.

96-well microplate를 이용한 Trichophyton Rubrum의 항진균제 감수성검사 (Antimycotic Susceptibility Testing of Trichophyton Rubrum by Microculture Method)

  • 이무웅;김종철;최종수;김기홍
    • Journal of Yeungnam Medical Science
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    • 제9권2호
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    • pp.396-406
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    • 1992
  • 저자들은 Granade와 Artis의 방법에 따라 96-well microplate와 24-well macroplate를 이 용하여 T. rubrum 9주를 대상으로 경구용 항진균제인 ketoconazole과 itraconazole에 대한 MIC를 측정하여 실제 임상사용 가능성을 알아보고 배양온도, 배양용기의 크기, 배지의 종류를 달리하여 MIC에 영향을 줄 수 있는 요소를 점검하여 다음과 같은 결론을 얻었다. 1. 96-well microplate를 사용하여 $25^{\circ}C$에서 배양시 균농도에 따른 판독시기의 차이는 높은 균농도(흡광도 2.0, 1.0)에서는 4일만에, 낮은 균농도(흡광도 0.5, 0.25)에서는 6-8일만에 판독할 수 있었고, MIC는 높은 균농도에서 높았으나 시간이 경과시 점차 차이가 줄어들었다. 2. $37^{\circ}C$$25^{\circ}C$에서 각각 배양시 배양온도에 따른 MIC의 차이는 96-well microplate를 사용하여 $37^{\circ}C$에서 배양시 ketoconazole에 대한 MIC는 0.006이하-$0.04{\mu}g/ml$, itraconazole에 대한 MIC는 0.006이하-$0.04{\mu}g/ml$였으며 $25^{\circ}C$에서의 ketoconazole에 대한 MIC는 0.08-$5.68{\mu}g/ml$, itraconazole에 대한 MIC는 0.006-$0.71{\mu}g/ml$$37^{\circ}C$에서의 MIC는 $25^{\circ}C$에서의 MIC에 비해 현저히 낮았다. 3. 24-well microplate와 96-well microplate에서 각각 배양시 배양용기의 크기에 따른 판독시기는 96-well microplate 액체배지에서는 4-6일로 24-well macroplate 액체배지에서의 8-12일에 비해 판독시기가 빨랐으나, MIC의 차이는 없었다. 4. 액체배지와 고체배지에서 배양시 배지종류에 따른 MIC의 차이는 액체배지를 함유한 24-well macroplate를 이용한 경우 ketoconazole에 대한 MIC는 0.006이하-$5.68{\mu}g/ml$, itraconazole에 대한 MIC는 0.006 이하-$0.36{\mu}g/ml$였고 고체배지에서는 ketoconazole에 대한 MIC는 0.006이하-$5.68{\mu}g/ml$, itraconazole에 대한 MIC는 0.006 이하-$5.68{\mu}g/ml$로 고체배지에서의 MIC가 다소 높게 측정되었다. 5. 이상의 결과를 종합하여 볼때 96-well microplate를 사용하여, 흡광도 1.0의 균농도로 접종하여, $25^{\circ}C$에서 배양 후 5-6일째 육안으로 판독하는 것이 항진균제 감수성 검사를 빠르고 간편하게 실시 할 수 있는 방법이다.

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Microplate hybridization assay for detection of isoniazid resistance in Mycobacterium tuberculosis

  • Han, Hye-Eun;Lee, In-Soo;Hwang, Joo-Hwan;Bang, Hye-Eun;Kim, Yeun;Cho, Sang-Nae;Kim, Tae-Ue;Lee, Hye-Young
    • BMB Reports
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    • 제42권2호
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    • pp.81-85
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    • 2009
  • Early and accurate detection of drug resistant Mycobacterium tuberculosis can improve both the treatment outcome and public health control of tuberculosis. A number of molecular-based techniques have been developed including ones using probe molecules that target drug resistance-related mutations. Although these techniques are highly specific and sensitive, mixed signals can be obtained when the drug resistant isolates are mixed with drug susceptible isolates. In order to overcome this problem, we developed a new drug susceptibility test (DST) for one of the most effective anti-tuberculosis drug, isoniazid. This technique employed a microplate hybridization assay that quantified signals from each probe molecule, and was evaluated using clinical isolates. The evaluation analysis clearly showed that the microplate hybridization assay was an accurate and rapid method that overcame the limitations of DST based on conventional molecular techniques.

생약 추출물에 의한 superoxide와 hydroxyl 라디칼 소거능 검색 방법의 개선 (An Improved method in Screening of Superoxide and Hydroxyl Radical Scavenging Activities of Plant Medicinal Extracts)

  • 이호섭;강대길
    • 생약학회지
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    • 제32권3호통권126호
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    • pp.253-256
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    • 2001
  • The present study was designed for the improvement of routine measurement of superoxide and hydroxyl radical scavenging activities utilized by a microplate reader. Superoxide radical scavenging activity by the ascorbic acid, which is a well-known superoxide scavenger, was determined in a dose-dependent manner. Hydroxyl radical scavenging activity by the thiourea, which is a well-known hydroxyl radical scavenger, was also well detected in a dose-dependent manner. Our results suggest that the use of microplate reader to assay the superoxide and hydroxyl radical scavenging activities improves the accuracy of data and enables the use of much smaller amounts of samples and/or reagents, with much simpler experimental procedure. Therefore, These methods appear to be suitable for screening of superoxide and hydroxyl radical scavenging activities in both the plant medicinal extracts and the isolated compounds.

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고추의 카로티노이드 함량 분석을 위한 신속·간편 마이크로플레이트법 (A Simple and Fast Microplate Method for Analysis of Carotenoids Content in Chili Pepper (Capsicum annuum L.))

  • 홍순철;한정헌;이준대;안율균;양은영;채수영;김수;윤재복
    • 원예과학기술지
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    • 제31권6호
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    • pp.807-812
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    • 2013
  • 본 연구에서는 폴리스틸렌 재질의 96-well 마이크로플레이트와 ELISA 판독기를 이용하여 고추의 카로티노이드 함량을 간편하게 분석할 수 있는 방법을 개발하고자 하였다. 분광광도계를 이용하지 않고 450nm 필터가 장착된 ELISA 판독기로도 카로티노이드 분석이 가능하였으나, 카로티노이드 추출 시 사용하는 아세톤으로 인해 마이크로플레이트의 표면이 불투명하게 변색되었다. 그러나 아세톤 색소추출물을 메탄올로 10배 희석한 용액을 사용할 경우, 플레이트 표면과 용액 내 카로티노이드 성분의 변화 없이 카로티노이드 측정이 가능하였고, ASTA-20.1법으로 측정된 결과와도 높은 상관관계를 보였다. 한편, 카로티노이드 추출 조건으로는 고춧가루 0.1g을 아세톤 10mL에 첨가하고 실온에서 4시간 동안 교반하는 것이 적합하였다. 본 실험의 ELISA 판독기를 이용한 색소분석법(Microplate법으로 명명)은 대량의 시료를 처리할 수 있을 뿐만 아니라 아세톤 폐액의 발생량을 ASTA-20.1법의 1/10 수준으로 줄일 수 있어 실제 고추 육종에서 고색소 계통을 선발할 때 매우 유용할 것으로 판단된다.

Nonlinear bending of multilayer functionally graded graphene-reinforced skew microplates under mechanical and thermal loads using FSDT and MCST: A study in large deformation

  • J. Jenabi;A.R. Nezamabadi;M. Karami Khorramabadi
    • Structural Engineering and Mechanics
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    • 제90권3호
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    • pp.219-232
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    • 2024
  • In current study, for the first time, Nonlinear Bending of a skew microplate made of a laminated composite strengthened with graphene nanosheets is investigated. A mixture of mechanical and thermal stresses is applied to the plate, and the reaction is analyzed using the First Shear Deformation Theory (FSDT). Since different percentages of graphene sheets are included in the multilayer structure of the composite, the characteristics of the composite are functionally graded throughout its thickness. Halpin-Tsai models are used to characterize mechanical qualities, whereas Schapery models are used to characterize thermal properties. The microplate's non-linear strain is first calculated by calculating the plate shear deformation and using the Green-Lagrange tensor and von Karman assumptions. Then the elements of the Couple and Cauchy stress tensors using the Modified Coupled Stress Theory (MCST) are derived. Next, using the Hamilton Principle, the microplate's governing equations and associated boundary conditions are calculated. The nonlinear differential equations are linearized by utilizing auxiliary variables in the nonlinear solution by applying the Frechet approach. The linearized equations are rectified via an iterative loop to precisely solve the problem. For this, the Differential Quadrature Method (DQM) is utilized, and the outcomes are shown for the basic support boundary condition. To ascertain the maximum values of microplate deflection for a range of circumstances-such as skew angles, volume fractions, configurations, temperatures, and length scales-a parametric analysis is carried out. To shed light on how the microplate behaves in these various circumstances, the resulting results are analyzed.