• Clinical and Experimental Reproductive Medicine
• /
• v.24 no.3
• /
• pp.335-340
• /
• 1997

Fluorescence in situ hybridization (FISH) techniques allow the enumeration of chromosome abnormalities and from a great potential for many clinical applications. In order to produce quantitative and reproducible results, expensive tools such as a cooled CCD camera and a computer software are required. We have developed a Chromosome Image Processing System (Chips) using FISH that allows the detection and mapping of the genetic aberrations. The aim of our study, therefore, is to evaluate the capabilities of our original system using a black-and-white video camera. As a model system, three repetitive DNA probes (D18Z1, DXZ1, and DYZ3) were hybridized to variety different clinical samples such as human metaphase spreads and interphase nuclei obtained from uncultured peripheral blood lymphocytes, uncultured amniocytes, and germ cells. The visualization of the FISH signals was performed using our system for image acquisition and pseudocoloring. FISH images were obtained by combining images from each of probes and DAPI counterstain captured separately. Using our original system, the aberrations of single or multiple chromosomes in a single hybridization experiment using chromosomes and interphase nuclei from a variety of cell types, including lymphocytes, amniocytes, sperm, and biopsied blastomeres, were enabled to evaluate. There were no differences in the image quality in accordance with FISH method, fluorochrome types, or different clinical samples. Always bright signals were detected using our system. Our system also yielded constant results. Our Chips would permit a level of performance of FISH analysis on metaphase chromosomes and interphase nuclei with unparalleled capabilities. Thus, it would be useful for clinical purposes.

• Journal of Microbiology and Biotechnology
• /
• v.31 no.9
• /
• pp.1323-1329
• /
• 2021

Micro-scale magnetic beads are widely used for isolation of proteins, DNA, and cells, leading to the development of in vitro diagnostics. Efficient isolation of target biomolecules is one of the keys to developing a simple and rapid point-of-care diagnostic. A zinc finger protein (ZFP) is a double-stranded (ds) DNA-binding domain, providing a useful scaffold for direct reading of the sequence information. Here, we utilized two engineered ZFPs (Stx2-268 and SEB-435) to detect the Shiga toxin (stx2) gene and the staphylococcal enterotoxin B (seb) gene present in foodborne pathogens, Escherichia coli O157 and Staphylococcus aureus, respectively. Engineered ZFPs are immobilized on a paramagnetic bead as a detection platform to efficiently isolate the target dsDNA-ZFP bound complex. The small paramagnetic beads provide a high surface area to volume ratio, allowing more ZFPs to be immobilized on the beads, which leads to increased target DNA detection. The fluorescence signal was measured upon ZFP binding to fluorophore-labeled target dsDNA. In this study, our system provided a detection limit of ≤ 60 fmol and demonstrated high specificity with multiplexing capability, suggesting a potential for development into a simple and reliable diagnostic for detecting multiple pathogens without target amplification.

• Proceedings of the Korean Vacuum Society Conference
• /
• 2013.08a
• /
• pp.281.2-281.2
• /
• 2013

Since heavy metal ions included in water or food resources have critical effects on human health, highly sensitive, rapid and selective analysis for heavy metal detection has been extensively explored by means of electrochemical, optical and colorimetric methods. For example, quantum dots (QDs), such as semiconductor QDs, have received enormous attention due to extraordinary optical properties including high fluorescence intensity and its narrow emission peaks, and have been utilized for heavy metal ion detection. However, the semiconductor QDs have a drawback of serious toxicity derived from cadmium, lead and other lethal elements, thereby limiting its application in the environmental screening system. On the other hand, Graphene oxide (GO) has proven its superlative properties of biocompatibility, unique photoluminescence (PL), good quenching efficiency and facile surface modification. Recently, the size of GO was controlled to a few nanometers, enhancing its optical properties to be applied for biological or chemical sensors. Interestingly, the presence of various oxygenous functional groups of GO contributes to opening the band gap of graphene, resulting in a unique PL emission pattern, and the control of the sp2 domain in the sp3 matrix of GO can tune the PL intensity as well as the PL emission wavelength. Herein, we reported a photoluminescent GO array on which heavy metal ion-specific DNA aptamers were immobilized, and sensitive and multiplex heavy metal ion detection was performed utilizing fluorescence resonance energy transfer (FRET) between the photoluminescent monolayered GO and the captured metal ion.

• Proceedings of the Korean Vacuum Society Conference
• /
• 2013.08a
• /
• pp.88-89
• /
• 2013

A variety of influenza A viruses from animal hosts are continuously prevalent throughout the world which cause human epidemics resulting millions of human infections and enormous industrial and economic damages. Thus, early diagnosis of such pathogen is of paramount importance for biomedical examination and public healthcare screening. To approach this issue, here we propose a fully integrated Rotary genetic analysis system, called Rotary Genetic Analyzer, for on-site detection of influenza A viruses with high speed. The Rotary Genetic Analyzer is made up of four parts including a disposable microchip, a servo motor for precise and high rate spinning of the chip, thermal blocks for temperature control, and a miniaturized optical fluorescence detector as shown Fig. 1. A thermal block made from duralumin is integrated with a film heater at the bottom and a resistance temperature detector (RTD) in the middle. For the efficient performance of RT-PCR, three thermal blocks are placed on the Rotary stage and the temperature of each block is corresponded to the thermal cycling, namely $95^{\circ}C$ (denature), $58^{\circ}C$ (annealing), and $72^{\circ}C$ (extension). Rotary RT-PCR was performed to amplify the target gene which was monitored by an optical fluorescent detector above the extension block. A disposable microdevice (10 cm diameter) consists of a solid-phase extraction based sample pretreatment unit, bead chamber, and 4 ${\mu}L$ of the PCR chamber as shown Fig. 2. The microchip is fabricated using a patterned polycarbonate (PC) sheet with 1 mm thickness and a PC film with 130 ${\mu}m$ thickness, which layers are thermally bonded at $138^{\circ}C$ using acetone vapour. Silicatreated microglass beads with 150~212 ${\mu}L$ diameter are introduced into the sample pretreatment chambers and held in place by weir structure for construction of solid-phase extraction system. Fig. 3 shows strobed images of sequential loading of three samples. Three samples were loaded into the reservoir simultaneously (Fig. 3A), then the influenza A H3N2 viral RNA sample was loaded at 5000 RPM for 10 sec (Fig. 3B). Washing buffer was followed at 5000 RPM for 5 min (Fig. 3C), and angular frequency was decreased to 100 RPM for siphon priming of PCR cocktail to the channel as shown in Figure 3D. Finally the PCR cocktail was loaded to the bead chamber at 2000 RPM for 10 sec, and then RPM was increased up to 5000 RPM for 1 min to obtain the as much as PCR cocktail containing the RNA template (Fig. 3E). In this system, the wastes from RNA samples and washing buffer were transported to the waste chamber, which is fully filled to the chamber with precise optimization. Then, the PCR cocktail was able to transport to the PCR chamber. Fig. 3F shows the final image of the sample pretreatment. PCR cocktail containing RNA template is successfully isolated from waste. To detect the influenza A H3N2 virus, the purified RNA with PCR cocktail in the PCR chamber was amplified by using performed the RNA capture on the proposed microdevice. The fluorescence images were described in Figure 4A at the 0, 40 cycles. The fluorescence signal (40 cycle) was drastically increased confirming the influenza A H3N2 virus. The real-time profiles were successfully obtained using the optical fluorescence detector as shown in Figure 4B. The Rotary PCR and off-chip PCR were compared with same amount of influenza A H3N2 virus. The Ct value of Rotary PCR was smaller than the off-chip PCR without contamination. The whole process of the sample pretreatment and RT-PCR could be accomplished in 30 min on the fully integrated Rotary Genetic Analyzer system. We have demonstrated a fully integrated and portable Rotary Genetic Analyzer for detection of the gene expression of influenza A virus, which has 'Sample-in-answer-out' capability including sample pretreatment, rotary amplification, and optical detection. Target gene amplification was real-time monitored using the integrated Rotary Genetic Analyzer system.

• Proceedings of the KIEE Conference
• /
• 2002.11a
• /
• pp.60-63
• /
• 2002

This paper presents bead-based microbiocihps to detect and separate target proteins. Micro beads coated with capture proteins were introduced into a microchamber, and target proteins flowing across the chamber were bound and concentrated. The chip was connected with an external fluid system. Bead surfaces were double-coated with photo-cleavable linkers and capture proteins. The proteins bound on the beads were photo-separated under UV irradiation, and excited to be measured in fluorescence. $38{\sim}50{\mu}m$ sized polystyrene beads were used. SOGs(silicon-on-glass) were used to fabricate the microchip having glasses bonded on both sides. 100 ${\mu}m$ thick silicon channel was formed through silicon deep RIE process. The upper glass cover had holed through to have inlets and outlets fabricated by powder-blastings. In this study, biotin and streptavidin were used as capture proteins and detection proteins, respectively. The protein mixtures of streptavidin, HSA(human serum albumin) and ovalbumin were applied for selective detection test.

• Toxicological Research
• /
• v.25 no.4
• /
• pp.189-193
• /
• 2009

Hypoxia inducible factor $1\alpha$ (HIF-$1\alpha$) is a potential marker of carcicnogenesis since it is overexpresssed in many human cancers such as brain, breast, and uterus, and its role has implicated in tumor cell growth and metastasis. In this study, we established a stable cell line that express green fluorescence protein (GFP)-fused hypoxia inducible factor $1\alpha$ (HIF-$1\alpha$) and evaluated the potential use of this cell line for assessment of carcinogenicity of chemical toxicants. Western blot analysis as well as fluorescence measurements showed that protein-level of GFP-HIF-$1\alpha$ was significantly enhanced in a dose-dependent manner upon treatment of hypoxia mimicking agents such as dexferrioxamine and $CoCl_2$. Well-Known tumor promoters such as mitomycin and methyl methanesulfonate. significantly induced the fluorescence intensity of GFP-HIF-$1\alpha$, whereas the known negative controls such as o-anthranilic acid and benzethonium chloride, did not. These results indicate that HIF-$1\alpha$ could be a biological parameter for detection of tumor initiators/promoters and suggest that the GFP-HIF-$1\alpha$ cell line is a useful system for screening of carcinogenic toxicants.

• Journal of Soil and Groundwater Environment
• /
• v.7 no.2
• /
• pp.35-44
• /
• 2002

In order to determine the contamination of the aromatic hydrocarbons in soil, a fiber-optic sensing technique with fluorescence detector has been proposed. Previous researches have shown that the optimal condition for detecting benzene, toluene, ethylbenzene, xylene (BTEX) was 260 nm /290 nm (excitation/emission wavelength). However, broader fluorescence spectra of BTEX-polluted soil sample ranging from 300 nm to 600 nm were observed. Additionally, the intensity of fluorescence increased with increasing BTEX concentration, which was conspicuous in the fine-particle soil, The overall results indicated that the suggested technique could be useful for in-situ monitoring system for subsurface oil-storage tank.

• Journal of the Korean Chemical Society
• /
• v.38 no.9
• /
• pp.653-659
• /
• 1994

The fluorescence intensities of europium and samarium can be greatly enhanced in the presence 2-thenoyltrifluoroacetone(TTA), n-octanol and Triton X-100 in aqueous solution of pH 7. It was also found that the fluorescence intensity can be greatly increased by the addition of excess of $La^{3+}$. The excitation and emission wavelengths of europium and samarium were 345 nm, 380 nm and 617 nm, 567 nm, respectively. The fluorescence intensity was a linear function of the concentration of europium and samarium in the range TEX>$1{\times}10^{-7}∼1{\tiems}10^{-9}\;M,\;1{\tiems}10^{-5}∼1{\times}10^{-7}\;M$, respectively, and the detection limits were 1$1{\times}10^{-11}\;M$ for europium and $1{\times}10^{-8}\;M$ for samarium and the luminescence mechanism of the system is discussed.

• Nuclear Engineering and Technology
• /
• v.55 no.10
• /
• pp.3844-3853
• /
• 2023

Although radiation and chemotherapy methods for cancer therapy have advanced significantly, surgical resection is still recommended for most cancers. Therefore, intraoperative imaging studies have emerged as a surgical tool for identifying tumor margins. Intraoperative imaging has been examined using conventional imaging devices, such as optical near-infrared probes, gamma probes, and ultrasound devices. However, each modality has its limitations, such as depth penetration and spatial resolution. To overcome these limitations, hybrid imaging modalities and tracer studies are being developed. In a previous study, a multi-modal laparoscope with silicon photo-multiplier (SiPM)-based gamma detection acquired a 1 s interval gamma image. However, improvements in the near-infrared fluorophore (NIRF) signal intensity and gamma image central defects are needed to further evaluate the usefulness of multi-modal systems. In this study, an attempt was made to change the NIRF image acquisition method and the SiPM-based gamma detector to improve the source detection ability and reduce the image acquisition time. The performance of the multi-modal system using a complementary metal oxide semiconductor and modified SiPM gamma detector was evaluated in a phantom test. In future studies, a multi-modal system will be further optimized for pilot preclinical studies.

• Journal of Microbiology and Biotechnology
• /
• v.27 no.4
• /
• pp.808-815
• /
• 2017

We demonstrated the quantitative detection of a toxin-producing Microcystis aeruginosa (M. aeruginosa) strain with the laboratory protocol of the NanoGene assay. The NanoGene assay was selected because its laboratory protocol is in the process of being transplanted into a portable system. The mcyD gene of M. aeruginosa was targeted and, as expected, its corresponding fluorescence signal was linearly proportional to the mcyD gene copy number. The sensitivity of the NanoGene assay for this purpose was validated using both dsDNA mcyD gene amplicons and genomic DNAs (gDNA). The limit of detection was determined to be 38 mcyD gene copies per reaction and 9 algal cells/ml water. The specificity of the assay was also demonstrated by the addition of gDNA extracted from environmental algae into the hybridization reaction. Detection of M. aeruginosa was performed in the environmental samples with environmentally relevant sensitivity (${\sim}10^5$ algal cells/ml) and specificity. As expected, M. aeruginosa were not detected in nonspecific environmental algal gDNA over the range of $2{\times}10^0$ to $2{\times}10^7$ algal cells/ml.