• Bulletin of the Korean Chemical Society
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• v.26 no.6
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• pp.979-982
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• 2005

• Bulletin of the Korean Chemical Society
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• v.26 no.2
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• pp.315-318
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• 2005

• Bulletin of the Korean Chemical Society
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• v.28 no.5
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• pp.783-790
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• 2007

The immobilization of proteins and their molecular interactions on various polymer-modified glass substrates [i.e. 3-aminopropyltriethoxysilane (APTS), 3-glycidoxypropyltrimethoxysilane (GPTS), poly (ethylene glycol) diacrylate (PEG-DA), chitosan (CHI), glutaraldehyde (GA), 3-(trichlorosilyl)propyl methacrylate (TPM), 3'-mercaptopropyltrimethoxysilane (MPTMS), glycidyl methacrylate (GMA) and poly-l-lysine (PL).] for potential applications in a nanoarray protein chip at the single-molecule level was evaluated using prismtype dual-color total internal reflection fluorescence microscopy (dual-color TIRFM). A dual-color TIRF microscope, which contained two individual laser beams and a single high-sensitivity camera, was used for the rapid and simultaneous dual-color detection of the interactions and colocalization of different proteins labeled with different fluorescent dyes such as Alexa Fluor® 488, Qdot® 525 and Alexa Fluor® 633. Most of the polymer-modified glass substrates showed good stability and a relative high signal-to-noise (S/N) ratio over a 40-day period after making the substrates. The GPTS/CHI/GA-modified glass substrate showed a 13.5-56.3% higher relative S/N ratio than the other substrates. 1% Top-Block in 10 mM phosphate buffered saline (pH 7.4) showed a 99.2% increase in the blocking effect of non-specific adsorption. These results show that dual-color TIRFM is a powerful methodology for detecting proteins at the single-molecule level with potential applications in nanoarray chips or nano-biosensors.

• Journal of the Korean Society of Visualization
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• v.3 no.1
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• pp.3-19
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• 2005

Novel optical techniques are presented for three-dimensional tracking of nanoparticles; Optical Serial Sectioning Microscopy (OSSM) and Ratiometric Total Internal Reflection Fluorescent Microscopy (R-TIRFM). OSSM measures optically diffracted particle images, the so-called Point Spread Function (PSF), and dotermines the defocusing or line-of-sight location of the imaged particle measured from the focal plane. The line-of-sight Brownian motion detection using the OSSM technique is proposed in lieu of the more cumbersome two-dimensional Brownian motion tracking on the imaging plane as a potentially more effective tool to nonintrusively map the temperature fields for nanoparticle suspension fluids. On the other hand, R-TIRFM is presented to experimentally examine the classic theory on the near-wall hindered Brownian diffusive motion. An evanescent wave field from the total internal reflection of a 488-nm bandwidth of an argon-ion laser is used to provide a thin illumination field of an order of a few hundred nanometers from the wall. The experimental results show good agreement with the lateral hindrance theory, but show discrepancies from the normal hindrance theory. It is conjectured that the discrepancies can be attributed to the additional hindering effects, including electrostatic and electro-osmotic interactions between the negatively charged tracer particles and the glass surface.

• Journal of the Korean Society of Visualization
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• v.8 no.4
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• pp.31-37
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• 2010

In this study, a novel hybrid micro/nano PIV system combining defocusing and TIRFM technique has been developed for the multiscale flow measurement. With the developed system, both far and near field velocity fields have been measured simultaneously in a 2D straight microchannel and the particle trajectories were extracted by the nearest tracking algorithm. The shear rate values taken from experimental results have been estimated by comparing with the analytical solution of 2D Poiseuille flow and it is confirmed that the result shows good agreement with the theoretical value.

• 한국가시화정보학회:학술대회논문집
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• 2004.12a
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• pp.91-110
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• 2004

A comprehensive three-dimensional nano-particle tracking technique in micro- and nano-scale spatial resolution using the Total Internal Reflection Fluorescence Microscope (TIRFM) is discussed. Evanescent waves from the total internal reflection of a 488nm argon-ion laser are used to measure the hindered Brownian diffusion within few hundred nanometers of a glass-water interface. 200-nm fluorescence-coated polystyrene spheres are used as tracers to achieve three-dimensional tracking within the near-wall penetration depth. A novel ratiometric imaging technique coupled with a neural network model is used to tag and track the tracer particles. This technique allows for the determination of the relative depth wise locations of the particles. This analysis, to our knowledge is the first such three-dimensional ratiometric nano-particle tracking velocimetry technique to be applied for measuring Brownian diffusion close to the wall.