• Title/Summary/Keyword: Patterned fluorescence images

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The First Synthesis of a Transition Metal-Catalyzed Homopolymer Having Pendent t-Boc-Protected Quinizarin for Patterned Fluorescence Images

  • Jimmy Yoo;Lee, Jae-Hyoung;Iwhan Cho;Ahn, Kwang-Duk;Kim, Jong-Man
    • Macromolecular Research
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    • v.11 no.1
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    • pp.69-72
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    • 2003
  • A homopolymer having pendent t-Boc-protected quinizarin moieties has been prepared for patterned fluorescence images. The homopolymer P(t-BQN (2-norbornenylmethyl di-tert-butoxycarbonylquinizarin)) 5 was prepared by palladium-catalyzed addition polymerization. The t-Boc-protecting groups of the polymer were efficiently removed during chemical amplification process and revealed original properties of quinizarin, allowing patterned fluorescence images in the polymer film.

Patterned Fluorescence Images with a t-Boc-Protected Coumarin Derivative

  • Min Sung-Jun;Park Bum Jun;Kim Jong-Man
    • Macromolecular Research
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    • v.12 no.6
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    • pp.615-617
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    • 2004
  • We have developed an efficient method for the generation of patterned fluorescence images using a protected precursor molecule. The t-Boc-protecting group of a coumarin derivative was readily removed from a polymer film upon irradiation with UV light in the presence of a photoacid generator to provide the original properties of the coumarin. Fine fluorescence patterns were obtained when using this photolithographic method.

Transient Protection of Intramolecular Hydrogen Bonding: A Simple but Elegant Approach for Functional Imaging

  • Kim, Jong-Man;Min, Sung-Jun;Park, Bum-Jun;Lee, Jae-Hyung;Ahn, Kwang-Duk
    • Macromolecular Research
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    • v.12 no.5
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    • pp.493-500
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    • 2004
  • We have developed a novel method for patterning functional images in thin polymer films. The key materials we utilized for the imaging were dihydroxyanthraquinones protected with acid-labile tert-butoxycarbonyl (t-Boc) blocking groups. Among the tested compounds, 1,4-dihydroxyanthraquinone (quinizarin; 1) underwent the most drastic change in terms of its color and fluorescence upon protection. We prepared the t-Boc-protected quinizarin and polymers bearing the protected quinizarins as pendent groups. To investigate the possibility of a single-component imaging system, we synthesized a styrenic monomer 14 incorporating protected quinizarin and a maleimide derivative 15 bearing a photoacid generating group and subjected them to polymerization. Selective removal of the protecting groups of the quinizarin moieties in the exposed area using photolithographic techniques allowed regeneration of quinizarin and patterned fluorescence images in the polymer films.

Radially patterned polycaprolactone nanofibers as an active wound dressing agent

  • Shin, Dongwoo;Kim, Min Sup;Yang, Chae Eun;Lee, Won Jai;Roh, Tai Suk;Baek, Wooyeol
    • Archives of Plastic Surgery
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    • v.46 no.5
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    • pp.399-404
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    • 2019
  • Background The objectives of this study were to design polycaprolactone nanofibers with a radial pattern using a modified electrospinning method and to evaluate the effect of radial nanofiber deposition on mechanical and biological properties compared to non-patterned samples. Methods Radially patterned polycaprolactone nanofibers were prepared with a modified electrospinning method and compared with randomly deposited nanofibers. The surface morphology of samples was observed under scanning electron microscopy (SEM). The tensile properties of nanofibrous mats were measured using a tabletop uniaxial testing machine. Fluorescence-stained human bone marrow stem cells were placed along the perimeter of the radially patterned and randomly deposited. Their migration toward the center was observed on days 1, 4, and 7, and quantitatively measured using ImageJ software. Results Overall, there were no statistically significant differences in mechanical properties between the two types of polycaprolactone nanofibrous mats. SEM images of the obtained samples suggested that the directionality of the nanofibers was toward the central area, regardless of where the nanofibers were located throughout the entire sample. Florescence images showed stronger fluorescence inside the circle in radially aligned nanofibers, with significant differences on days 4 and 7, indicating that migration was quicker along radially aligned nanofibers than along randomly deposited nanofibers. Conclusions In this study, we successfully used modified electrospinning to fabricate radially aligned nanofibers with similar mechanical properties to those of conventional randomly aligned nanofibers. In addition, we observed faster migration along radially aligned nanofibers than along randomly deposited nanofibers. Collectively, the radially aligned nanofibers may have the potential for tissue regeneration in combination with stem cells.

Construction and Patterning of the biospecific affinity surfaces on gold using dendrimer

  • Hong, Mi-Yeong;Yun, Hyeon-Cheol;Kim, Hak-Seong
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.703-706
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    • 2000
  • We constructed a biospecific affinity surface using hyper-branched dendrimers on gold for biospecific recognition, and characterized the resulting surfaces by using confocal fluorescence microscopy. The dendrimer monolayer was firstly constructed on the mercaptoundecanoic acid SAM/Au with pentafluorophenyl ester activation and further functionalized with sulfo-NHS-biotin, an activated ester of biotin. To confirm the formation of biospecific affinity surface, FITC(fluorescein isothiocyanate)-labeled avidin was loaded onto the biotinylated dendrimer monolayer, and fluorescence images of the bound avidins were investigated with a confocal microscope. The constructed biospecific affinity surface showed a much more dense and uniform fluorescence compared to those from poly-L-lysine- and cystamine SAM-based affinity surfaces. For the dependency on the concentration of added FITC-labeled avidin on the affinity surface, derived fluorescence could be detectable from as low as $1{\mu}g/ml$, and intensified up to $50{\mu}g/ml$. Further reaction of FITC-labeled avidin layer with TMR(tetramethylrhodamine)-biocytins resulted in the efficient FRET(fluorescence resonance energy transfer) phenomenon. As an extension of the study, we attempted a patterning of the affinity surfaces on gold by microcontact printing. Fluorescence of the patterned surface demonstrated that FITC-labeled avidin molecules were specifically bound to the biotinylated patches.

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Integrated Rotary Genetic Analysis Microsystem for Influenza A Virus Detection

  • Jung, Jae Hwan;Park, Byung Hyun;Choi, Seok Jin;Seo, Tae Seok
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.08a
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    • pp.88-89
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    • 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.

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