Earticle

본 연구에서는 sonificator를 장착하여 세포막을 파쇄하고 현장에서 형광을 이용하여 조작이 간편하고 단시간에 DNA를 측정할 수 있는 자동화된 형광기를 개발하기 위하여 그람 양성균으로 간균인 Bacillus globigii와 구균인 Streptococcus epidermidis를 대상으로 최적의 세포 파쇄조건을 확립하고자 하였다. Sonificator probe 직경에 따라 세포의 파쇄 효과가 큰 차이를 보였으며 13 ㎜ probe로 20초동안 sonification시키는 것이 가장 효율적인 것으로 나타났으며 간균인 Bacillus globigii가 구균인 Streptococcus epidermidis보다 더 파쇄가 잘 되는 것으로 나타났다. 시료에 잠긴 Sonificator probe tip 깊이에 따라서도 세포의 파쇄 효과가 크게 나타났는데 시료에 잠긴 probe tip의 깊이가 깊을수록 큰 파쇄 효과를 발휘하였다. 선정된 최적의 파쇄 조건에서 최저 탐지농도는 Bacillus globigii, Streptococcus epidermidis 모두 5 × 105 CFU/㎖의 농도를 탐지 가능한 것으로 나타났다.

This study was carried out to establish the optimum condition for cell disruption with a sonificator in the detection of the gram positive bacteria, Bacillus globigii and Streptococcus epidermidis for the purpose of developing automatic fluorometer. The efficiency of sonificator on the Bacillus globigii and Streptococcus epidermidis disruption differed greatly according to the diameter of sonificator probe tip. The larger sonificator probe diameter showed greater disruption. Bacillus globigii was more disruptive than Streptococcus epidermidis. Sonificator probe of the 13 mm diameter was the most efficient one when sample was sonificated for 20 seconds. The detection limits of Bacillus globigii and Streptococcus epidermidis were 105 CFU/㎖ and 5 × 105 CFU/㎖ respectively when samples were sonificated for 20 seconds with a sonificator probe of 13 mm diameter.

본 연구에서는 sonificator를 장착하여 세포막을 파쇄하고 현장에서 형광을 이용하여 조작이 간편하고 단시간에 DNA를 측정할 수 있는 자동화된 형광기를 개발하기 위하여 그람 음성균인 Escherchia coli를 대상으로 최적의 세포파쇄조건을 확립하고자 하였다. Incubation time은 형광량에 크게 영향을 주지 못하는 것으로 나타났으며, 가열처리 방법은 현장에서 세포를 파괴하는 방법으로는 파쇄효과가 미약하고 적합하지 않은 것으로 나타났다. Sonificator probe 직경에 따라 세포의 파쇄 효과가 큰 차이를 보였으며 13 ㎜ probe로 20초 동안 sonification시키는 것이 가장 효율적인 것으로 나타났다. 시료에 잠긴 Sonificator probe tip 깊이에 따라서도 세포의 파쇄 효과가 크게 나타났는데 시료에 잠긴 probe tip의 깊이가 깊을수록 큰 파쇄 효과를 발휘하였다. 선정된 최적의 파쇄 조건에서 5 × 105 CFU/㎖의 Escherchia coli를 탐지 가능한 것으로 나타났다.

This study was carried out to establish the optimum condition for cell disruption with a sonificator in the detection of the gram negative bacteria, E. coli for the purpose of developing automatic fluorometer. The efficiency of sonification on the E. coli disruption was greatly dependent on the diameter of sonificator probe tip. The larger sonificator probe diameter showed greater disruption effect. Sonificator probe of 13 mm diameter was the most efficient one for E. coli when sonificated for 20 seconds. The efficiency of the E. coli disruption differed greatly according to the depth of sonificator probe tip sank in the sample solution. The shorter the distance between probe tip end and the bottom of the container, the higher the disruption efficiency. The detection limit of E. coli was 5 × 105 CFU/㎖ when sample was sonificated for 20 seconds with a sonificator probe of 13 mm diameter.

대기중 가스상 $H_2O_2$(Hydrogen Peroxide)는 액상 화학반응과 기상 라디칼반응사이에 연결고리의 역할을 할 뿐만 아니라, 대기중의 $SO_2$를 $H_2SO_4$로 산화시키는 산화제로서 구름, 안개, 이슬 및 빗물의 산화에 중요한 역할을 담당한다. 또한 가스상 $H_2O_2$는 연쇄종결자와 $HO_2$.(hydroperoxyl radical)농도의 지표로서 광화학 스모그에 있어 중요한 화학종이기도 하다. $H_2O_2$농도의 증가는 결국 대기의 산화율 및 속도를 증가시키고 대류권내의 액상중에서 $H_2SO_4$ 생성을 가속화시킨다는 것은 이미 잘 알려져 있는 사실이다. (중략)

A miniaturized fluorescence detection system based on total internal reflection (TIR) configuration, which is applicable to detecting the presence of biological materials labeled with fluorescence dye in micro total analysis systems (${\mu}TAS$), is proposed. In conventional fluorescence testing and analysis devices, interference between the excitation light beam and the emitted light from dyes is unavoidable. This paper presents a fluorescence detection system based on TIR configuration that allows the excitation light beam and the emitted light to be spatially perpendicular to each other so as to minimize the interference where fluorescence emission is detected at the orthogonal angle to the excitation beam. We achieved the limit of detection of about 5 nmol/L with a high linearity of 0.994 over a wide range of 6-FAM mol concentration, being comparable to that in earlier studies.

Quinine quantitative analysis, photo-bleaching, fluorescence quenching, and urine analysis have been performed by means of a UV-LED fluorometer, which can be easily built and used in a high-school laboratory. The quinine detection range is estimated to be 0.05-80 ppm, enough for many classroom luminescence experiments. The quinine content in commercial tonic water is determined from the calibration curve, and UV photo-bleaching of this anti-malarial drug is demonstrated with clear wavelength dependence. Halide quenching of quinine fluorescence is also observed and the increase in quenching efficiency in the order of $Cl^-$, $Br^-$, and $I^-$ is evident. Urine analyses for the student volunteers have been carried out and the results clearly reveal excretion of the ingested quinine. The student participants are exuberant throughout the course of this study and sense the practices resourceful.

In this study, the PHYTO-PAM-fluorometric method was used to evaluate the ETR$_{max}$ in terms of sensitivity to DIN/DIP against 14 microalgae: Prorocentrum micans, Heterocapsa triquetra, Gymnodinium impudicum, Cymnodinium catenatum, Amphidinium caterae, Chlorella vulgaris, Chroococcus minutus, Microcystis aeruginosa, Chlorella ellipsoidea, Nannochloris oculata, Oocystis lacustris, Chroomonas salina, Gloeocystis gigas, and Prymnessium parvum. We found that P. micans, H. triquetra, and A. caterae exposed to the maximum level of DIN/DIP were significantly smaller in the ETR$_{max}$ than that of the minimum and moderate mixture. Unlikely the ETR$_{max}$, the initial slope alpha was not significantly different at the level of 60 DIN/DIP. In G. catenatum, the moderate levels of 15 and 20 in DIN/DIP were found to be significantly different from the ETR$_{max}$ at Chl-Ch4. Gymnodinium impudicum had a higher value than that of the ETR$_{max}$ than that of dinoflagellates used in this study, ranging from 306.1 (Ch4, DIN/DIP: 10) to 520.1 (Ch4, DIN/DIP: 30). The ETR$_{max}$ value obtained from other microalgae was similar to C. impudicum at any of the ratios of DIN/DIP and channels. Consequently, the influence of offshore water current assures us of the suppression of photosynthesis and electron transport rate in dinoflagellates. Gymnodinium impudicum has not been researched in the area of red tides in Korea, but it will be enough to creat the massive algal blooms in the future because of higher potential photochemical availability.