This work involves the development of a novel waste-derived carbon dots (CDs) conjugated with silver (Ag) nanohybrid system-based Fluorescence Resonance Energy Transfer (FRET) sensor for the detection of melamine. CDs and Ag nanoparticles served as energy donors and energy acceptors, respectively. CDs were synthesized from orange peel waste through a combined hydrothermal and ultra-sonication route. The synthesized CDs had hydroxyl, amino, and carboxyl groups on their surface, explaining that waste-derived CDs can act as reducing and stabilizing agents and showed strong absorption and fluorescence emission at 305 and 460 nm, respectively. The bandgap, linear refractive index, conduction band, and valance band potential of CDs were observed to be 2.86, 1.849, 1.14, and 4.002 eV, respectively. No significant difference was observed in the fluorescence properties at different pH (acid and alkaline) and ionic concentrations. Given their fluorescent nature, the synthesized CDs were used for the detection of melamine. The fluorescence of CDs was found to be quenched by Ag+ due to the FRET energy transfer between CDs to Ag. Notably, the zeta potential of Ag@CDs was changed from − 28.7 mV to − 30.6 mV after the incorporation of Ag+. Ag@CDs showed excellent selectivity and sensitivity toward the sensing of melamine in the aqueous solutions with the limit of detection ~ 0.85 μM. Increasing the melamine level also raises the FL intensity of Ag@CDs. The substrate was effectively used in the detection of melamine in milk as a real application and the recovery percentage was found to be 98.03%. Moreover, other adulterants such as urea and formaldehyde can be detected selectively by Ag@CDs. Overall, the synthesized Ag@CDs can be used as an efficient material for sensing applications involving such food adulterants.
We successfully synthesize water-dispersible CTAB-capped CdSe@ZnS quantum dots with the crystal size of the CdSe quantum dots controlled from green to orange colors. The quenching effect of Fe(DTC)3 is very efficient to turn off the emission light of quantum dots at four molar ratios of the CdSe quantum dots, that is, the effective covering the surface of quantum dots with Fe(DTC)3. However, the reaction with Fe(DTC)3 for more than 24 h is required to completely realize the quenching effect. The highly quenched quantum dots efficiently detect nitric oxide at nano-molar concentration of 110nM of NO with 34% of recovery of emission light intensity. We suggest that Fe(DTC)3-hybridized CdSe@ZnS quantum dots are an excellent fluorescence resonance energy transfer probe for the detection of nitric oxide in biological systems.
The purpose of this study was to examine the characteristics of acetaminophen (APAP)-induced liver damage, using fluorescence bioimaging, serum biochemistry, and histopathology. At six weeks of age, eighteen mice were divided into three groups as group 1 (G1) as control, group 2 (G2) as fluorescence probe control and group 3 (G3) as APAP-treated. G3 mice were orally treated with APAP (800 mg/kg b.w.), while G1 and G2 mice were treated with 0.9% saline. Twenty-two hours after APAP treatment, G2 and G3 mice were intravenously treated with Annexin-Vivo 750 as probe, while G1 mice were treated with saline. Fluorescence bioimaging was performed at two hours after probe treatment. The mice were sacrificed and serum levels of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase and lactate dehydrogenase were analyzed. Liver damage was examined by hematoxylin and eosin (H&E) staining and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining. In vivo bioimaging, fluorescence intensity of the region of interest (ROI) was significantly increased in the livers of G2 and G3 mice compared with those in G1 mice (p<0.05 and p<0.01). In addition, ex vivo bioimaging confirmed that the fluorescence intensity of the ROI was significantly increased in the livers of G2 and G3 mice compared with those in G1 mice (p<0.05 and p<0.01). All examined serum parameters of G3 were significantly increased compared with G1 and G2 (p<0.05 and p<0.01). H&E examination showed acute hepatic cell necrosis in the livers of G3 mice, while there was no cell death in the livers of G1 and G2 mice. TUNEL staining also showed many cell death features in G3 mice, whereas no pathological findings were shown in G1 or G2 mice. In summary, fluorescence bioimaging showed the possibility of cell death detection in the livers of mice treated with APAP, and this was corroborated by serum chemistry and histopathological examination.
본 연구는 육안판단이 아닌 엽록소형광 이미지 측정기 법을 이용하여 비파괴적으로 수박접목묘 플러그트레이 단일 셀에 대해 건조스트레스를 정량화하고자 수행되었 다. 접목 후 6일차 수박접목묘를 3일동안 균일한 관수관 리 하에서 재배한 후 건조스트레스를 부여하였다. 이후 플러그트레이 단일 셀 형태의 수분함량센서를 이용하여 D1(53.0%, 충분한 수분상태)단계부터 D9(15.7%, 극심한 건조스트레스)단계까지 9개 그룹으로 분류하고 엽록소 형광을 측정하였다. 또한 건조스트레스에 영향을 받은 묘(D5-D9)에 재관수하여 육안판단으로 확인되지 않은 광합성 및 생육 회복 수준을 측정하였다. 3개의 건조스트레스 단계의 엽록소형광 곡선 형태는 건조스트레스 조기 탐지에 대해 다른 양상을 보였다. 총 16개의 엽록소 형광 지수는 건조스트레스에 노출되면서 지속적으로 감소하였으며, 육안으로 판단 가능한 D5(32.1%)단계에서 크게 감소하였다. 형광감소율(Rfd_Lss)는 초기 건조스트 레스 수준(D5-D6)에서 명확하게 감소하기 시작하였으며, 최대 광화학효율(Fv/Fm)은 극심한 건조스트레스 수준 (D7-D9)에서 크게 감소하였다. 따라서, Rfd_Lss 및 Fv/ Fm 지수를 건조스트레스의 초기 및 이후 단계에서 생육 및 광합성 회복 평가를 위한 지표로 선정하였다. 개별 엽록소형광 지수의 수치값 차이와 엽록소형광 이미지를 통해 건조스트레스 수준이 직관적으로 확인되었다. 이러 한 결과는 Rfd_Lss와 Fv/Fm은 각각 초기 및 극심한 건 조스트레스를 탐지하지 위한 엽록소형광 지수로 활용될 수 있으며, Fv/Fm은 재관수시 회복 평가를 위한 최적의 엽록소형광 지수로 판단된다.
In this study, we have developed a fluorescence chromatographic assay for the quantification of total cholesterol in serum, which is a well-known risk predictor for cardiovascular diseases. The new assay system consists of a chromatographic strip in a cartridge, enzyme buffer containing cholesterol esterase, cholesterol oxidase, horseradish peroxidase, and color developer AEC, and a laser fluorescence scanner. The correlation coefficient (r) between cholesterol concentration and relative fluorescence units was 0.968 in the new assay, showing a reliable linearity through the tested range of cholesterol. Recovery test and comparability with a Hitachi 747 instrument showed 106.5-94% and r = 0.939 (p<0.001), respectively. The new assay system for cholesterol was developed as a pre-POCT platform conducted in clinics since it is fast (8 min) and uses a small volume of sample (5 μl), and it may be applied for on-site diagnostics to replace expensive automated biochemical analyzer.
본 논문에서는 식품 중 퀴놀론계(QNs) 합성항균제(marbofloxacin, norfloxacin, danofloxacin, ciprofloxacin, enrofloxacin, difloxacin, sarafloxacin, oxolinic acid 그리고 flumequine) 9종을 분석하기 위하여, 액-액 추출 과정을 거쳐서 형광검출기가 장착된 액체크로마토그래피를 이용하여 QNs를 효율적으로 동시분석하는 방법을 확립하였다. 컬럼은 Zorbax Eclipse XDB-C8(150 mm×4.6 mm, 5 μm), 이동상 용매는 200 mM ammonium acetate buffer (pH 4.5)와 ACN로 기울기 용리를 사용하였으며, 유속은 1.5 ml/min, 그리고, 주입량은 10 μl로 설정하여 분석하였다. 확립 된 분석조건으로, 표준검정 곡선은 10-500 μg/kg의 농도범위에서 상관계수가 0.9989 이상의 양호한 직선성을 나타내었으며, 회수율은 50, 100 그리고 500 μg/kg의 농도에서 89.6-106.5 %로, 향상된 추출효율을 나타내었다. 검출한계는 1-16 μg/kg이었고, 정량한계는 3-47 μg/kg이었으며, 일내(intra-day)와 일간(inter-day) 정밀도(CV%)는 0.2-15.0 %, 0.5-11.7 %이었다. 따라서, 확립된 분석방법은 광어 및 계란 중의 QNs을 효과적으로 분석하는데 이용될 수 있을 것으로 사료된다.
HPLC에 의한 주요 aflatoxins(afatoxin B₁, B₂, G₁ 및 G₂)의 동시 분석에서 postcolumn 유도체화법을 시도하였다. Electrochemical cell(Kobra-cell)을 사용한 postcolumn 유도체화법은 기존의 precolumn 유도체화법보다 분석시간을 단축하였으며(약 1/2 단축), 더 안전하고, 향상된 분석능을 보였다. Aflatoxin B₁과 G₁의 경우 10~100 ppb에서, 그리고 B₂와 G₂의 경우 3~30 ppb에서 직선성을 나타내었다. Aflatoxin B₁과 G₁은 각각 88.9% 및 100.5%로 양호환 회수육을 보였다. Aflatoxin B₂와 G₂의 경우 분리도는 우수하였으나 회수율에 있어서 변이가 크게 나타났다.
A new chemosensor based on a self-assembled system has been devised to detect Hg2+ions efficiently. We demonstrated that the amphiphilic building blocks consisting of pyrene and boronic acid (1) aggregate in aqueous solutions and provide an outstanding sensing platform for sensitive detection. The self-assembled 1 exhibited high selectivity and sensitivity for Hg2+ion detection via fluorescence quenching, where the Hg2+ion detection ensued from a fast transmetallation of 1. The Stern-Volmer (SV) quenching constant for its fluorescence quenching by Hg2+ions was approximately 1.58 × 108 M-1. In addition, self-assembled 1 exhibited excellent sensing abilities at nano-molar concentration levels when tap water and freshwater samples were contaminated with of Hg2+ ions.
Single nucleotide polymorphisms (SNPs) are valuable DNA markers due to their abundance and potential for use in automated high-throughput genotyping. Numerous SNP genotyping assays have been developed. In this report, one of effective and high throughput SNP genotyping assays, which was named the template-directed dye-terminator incorporation with fluorescence polarization detection (FP-TDI) was described. Although the most of this assay succeed, the objective of this work was to determine the reasons for the failures, find ways to improve the assay and reduce the running cost. Ninety F2 -derived soybean, Glycine max (L.) Merr., RILs from a cross between 'Pureunkong' and 'Jinpumkong 2' were genotyped at four SNPs. FP measurement was done on Victot3 microplate reader (perkinelmer Inc., Boston, MA, USA). Increasing the number of thermal cycles in the single-base extension step increased the separation of the FP values between the products corresponding to different genotypes. But in some assays, excess of heterozygous genotypes was observed with increase of PCR cycles. We discovered that the excess heterozygous was due to misincorporation of one of the dyeterminators during the primer extension reaction. After pyrophosphatase incubation and thermal cycle control, misincoporation can be effectively prevented. Using long amplicons instead of short amplicons for SNP genotyping and decreasing the amount of dye terminator and Acyclopol Taq polymerase to 1/2 or 1/3 decreased the cost of the assay. With these minor adjustments, the FP-TDI assay can be used more accurately and cost-effectively.