• Journal of the Korean Chemical Society
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• v.67 no.1
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• pp.64-67
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• 2023

• Molecules and Cells
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• v.26 no.1
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• pp.87-92
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• 2008

We employed dual color Fluorescence Cross Correlation Spectroscopy (FCCS) to measure the interaction between PKA regulatory (RII) and catalytic subunits (CAT) in living cells. Elevation of intracellular cAMP with forskolin decreased the cross-correlation amplitude between RFP-fused RII (RII -mRFP) and GFP-fused CAT (CAT-EGFP) by 50%, indicating that cAMP elevation leads to dissociation of RII-CAT complexes. Moreover, diffusion coefficient analysis showed that the diffusion rate of CAT-EGFP was significantly increased, suggesting that the decreased RII-CAT association caused by cAMP generated free CAT subunits. Our study demonstrates that in vivo FCCS measurements and their quantitative analysis permit one not only to directly quantify protein-protein interactions but also to estimate changes in the intracellular cAMP concentration.

• Korean Journal of Optics and Photonics
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• v.24 no.2
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• pp.71-76
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• 2013

Using fluorescence correlation spectroscopy, we analyzed the change of effective focal volume of a confocal system with light intensity. The fluorescence correlation spectroscopy system was home-built in accordance with the He-Ne laser with a wavelength of 632.8 nm, and two kinds of samples (AlexaFluor657 and Quantum dot655) suitable for the wavelength of the laser beam were used. For each sample, we analyzed and compared the correlation functions obtained while changing the intensity of the light source in a range of 1~50 ${\mu}W$. The result shows that the radius of the focal area increases linearly through the increase of particle number and diffusion time in response to an intensity change in weak light below 10 ${\mu}W$. In the higher intensity region (>10~15 ${\mu}W$), the increasing rate of particle number and diffusion time keep increasing but at a much slower rate. Through this result, it was also found that the radius increasing rate of the focal area was reduced however, the radius still increased slightly.

• Korean Journal of Optics and Photonics
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• v.23 no.3
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• pp.113-118
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• 2012

The concentration and hydrodynamic radius of nano-sized fluorescence particles diffusing in solution were compared by using fluorescence correlation spectroscopy (FCS), which can measure the variation of the correlation function of a fluorescence signal by size and number of particles. The used nano-sized fluorescence particles are Alex Fluor 647, quantum dots, and fluorescence beads, and three kinds of sample solutions with different concentrations were prepared by dilution to 1/10 and 1/100 with distilled water for each kind of particles. The effective focal volumes were calculated by using the known diffusion coefficient of Alexa Fluor 647 particles, and the diffusion time, number of particles in focal volume, and variation of concentration according to the dilution could be measured by the FCS system. Through this study, we determined that the concentrations of arbitrarily diluted sample solutions can be measured by a home-built FCS setup in the range of 0.1 nM ~ 10 nM and that the diffusion coefficient of the quantum dot was $27{\pm}1{\mu}m^2/s$.

• Journal of the Optical Society of Korea
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• v.12 no.3
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• pp.170-173
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• 2008

In this study, we simulated a statistical model of FCS(fluorescence correlation spectroscopy) based on a Poisson process to understand and explain observations of the experiment performed on molecules of ultra-low concentration by the home-built laser-scanning confocal microscope. The statistical model confirmed that the relative mean square amplitude of fluctuations is shown to be inversely proportional to the average number of molecules, even in the ultra-low concentration, if some conditions are satisfied. Signal-to-noise ratio and the variability of dwelling time under the confocal volume were found to be effective conditions for the experiment.

• Journal of the Optical Society of Korea
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• v.18 no.5
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• pp.598-604
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• 2014

A polarization fluorescence correlation spectroscopy system based on a confocal microscope was built to study the rotational and translational diffusion of CdSe/CdS quantum rods (Q-rods), with the same and different polarization states between the polarizer and the analyzer (i.e. the XXX and XYY states). The rotational diffusion amplitude showed the dependences on polarization of $0.75{\pm}0.05$ in the XXX state and $0.26{\pm}0.03$ in the XYY state, when the translational diffusion amplitude was 1. The diffusion coefficients of the Q-rods were found based on their translational and rotational diffusion times in the two polarization states, in solutions with viscosity ranging from 0.9 to 6.9 cP. The translational and rotational diffusion coefficients ranged from $1.5{\times}10^{-11}$ to $2.6{\times}10^{-12}m^2s^{-1}$ and from $2.9{\times}10^5$ to $5.6{\times}10^4s^{-1}$, respectively.

• Journal of the Korean Chemical Society
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• v.55 no.5
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• pp.746-750
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• 2011

Fluorescence correlation spectroscopy (FCS) is an emerging fluorescence technique used to study the dynamics of proteins on a millisecond to microsecond time scale at the single-molecule level. Solution pH-modulated protein conformational changes can be manifested by binding rate, fluorescence lifetime, and binding specificity of a probe molecule. The fluorescence lifetime of Nile red (NR) bound to apomyoglobin (apoMb) was measured to be $6{\pm}0.3$ ns, much longer than that in water solution ($2.9{\pm}0.2$ ns). As the unfolding population of apoMb increased by lowering pH of solution, the fraction for the longer lifetime of NR decreased with an increasing fraction for the shorter lifetime of NR in water. Unlike 1-anilino-8-naphthalene sulfonic acid, which has many lifetimes due to nonspecific binding to the unfolded apoMb, NR bound to apoMb possesses only a single lifetime. These results suggest that NR binds specifically to native apoMb and thus can be utilized to probe the folding/unfolding dynamics of apoMb using FCS.

• Journal of the Korean Chemical Society
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• v.47 no.5
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• pp.447-459
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• 2003

Generalized 2D correlation spectroscopy has been applied extensively to the analysis of spectral data sets obtained during the observation of a system under some external perturbation. It is used in various fields of spectroscopy including IR, Raman, UV, fluorescence, X-ray diffraction, and X-ray absorption spectroscopy (XAS) as well as chromatography. 2D hetero-spectral correlation analysis compares two completely different types of spectra obtained for a system under the same perturbation. Because of the wide range of applications of this technique, it has become one of the standard analytical techniques for the analytical chemistry, physical chemistry, biochemistry, and so on, and for studies of polymers, biomolecules, nanomaterials, etc. In this paper, we will introduce the principle of generalized 2D correlation spectroscopy and its applications that we have studied.

• Journal of Biosystems Engineering
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• v.35 no.2
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• pp.124-131
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• 2010

Chlorophyll fluorescence has been researched for assessing fruit post-harvest quality and condition. The objective of this preliminary research was to investigate the potential of fluorescence spectroscopy for measuring apple fruit quality. Ultraviolet (UV) and blue light was used as an excitation source for inducing fluorescence in apples. Fluorescence spectra were measured from 'Golden Delicious' (GD) and 'Red Delicious' (RD) apples using a visible/near-infrared spectrometer after one, three, and five minutes of continuous UV/blue light illumination. Standard destructive tests were performed to measure fruit firmness, skin and flesh color, soluble solids and acid content from the apples. Calibration models for each of the three illumination time periods were developed to predict fruit quality indexes. The results showed that fluorescence emission decreased steadily during the first three minutes of UV/blue light illumination and was stable within five minutes. The differences were minimal in the model prediction results based on fluorescence data at one, three or five minutes of illumination. Overall, better predictions were obtained for apple skin chroma and hue and flesh hue with values for the correlation coefficient of validation between 0.80 and 0.90 for both GD and RD. Relatively poor predictions were obtained for fruit firmness, soluble solids content, titrational acid, and flesh chroma. This research has demonstrated that fluorescence spectroscopy is potentially useful for assessing selected quality attributes of apple fruit and further research is needed to improve fluorescence measurements so that better predictions of fruit quality can be achieved.

• Nuclear Engineering and Technology
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• v.32 no.5
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• pp.446-456
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• 2000

Laser induced photoacoustic, fluorescence, and photon correlation spectroscopies were applied to the chemical characterization of radionuclides in connection with the radiowaste treatment and disposal. Their measuring principles and systems were briefly described together with their advantages over conventional spectroscopies. Also, other applications of lasers are introduced. Laser induced photoacoustic spectra were measured for a P $r^{3+}$ solution with a very low molar absorptivity. The detection sensitivity was 4.3 $\times$10$^{-5}$ c $m^{1}$ and was 100 times better than that of a UV/VIS spectrophotometer. The Eu(III) excitation spectra($^{7}$ $F_{0}$ longrightarrow $^{5}$ $D_{0}$ transition) were measured for Eu(III)-phthalate complexes using laser fluorescence spectroscopy, showing that only two species, 1:1 and 1:2 complexes, are present in the Eu(III)-phthalic acid system. The size and size distribution for colloidal humic acids and Eu(III)-humate colloids was determined using photon correlation spectroscopy. The presence of Eu(III) enhanced the aggregation of humic acids.s.