• 대한화학회지
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• 제67권1호
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• pp.64-67
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• 2023

• Molecules and Cells
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• 제26권1호
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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.

• 한국광학회지
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• 제24권2호
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• pp.71-76
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• 2013

형광상관분광법을 이용하여 광세기에 따른 공초점 시스템의 유효 초점 부피의 변화를 분석하였다. 형광상관분광장치는 632.8 nm 파장의 He-Ne 레이저에 맞춰서 실험실에서 자체 제작하였고, 시료 또한 레이저 파장에 적합한 두 종류의 시료 AlexaFluor647과 quantum dot 655를 사용하였다. 각 시료에 대해 광원의 세기를 1~50 ${\mu}W$ 범위내에서 변화시켜가며 얻어진 상관함수를 비교 분석하였다. 10 ${\mu}W$ 이하의 약한 광 세기에서는 세기 변화에 따라 입자수와 확산시간이 증가하는 것을 통해 초점 영역의 반지름이 선형적으로 증가하는 결과를 보였다. 반면 10~15 ${\mu}W$ 이상에서는 입자의 수와 확산 시간의 증가율은 감소하였지만 미세하게 계속해서 증가하는 결과를 보였고, 이 결과를 통해 초점영역의 반지름 역시 증가율은 감소하였지만 미세하게 증가한 것을 알 수 있었다.

• 한국광학회지
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• 제23권3호
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• pp.113-118
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• 2012

형광입자의 크기와 수에 따라 형광 신호의 상관함수 변화를 측정하는 형광상관분광법을 이용하여 용액 내에서 확산 운동하는 나노크기 형광 입자들의 농도와 유체역학적 반지름을 비교하였다. 시료에 사용된 나노크기 형광 입자들은 Alexa Fluor 647, 양자점, 형광 bead이고, 증류수에서 1/10, 1/100로 입자들이 들어있는 용액을 희석하여 각 입자들에 대해 3가지의 다른 농도의 시료를 준비하였다. Alex Fluor 647의 알려져 있는 확산시간을 이용하여 형광상관분광장치의 유효초점 부피를 구하고, 각 입자들의 확산계수, 크기, 희석에 따른 농도 변화를 측정할 수 있었다. 본 연구를 통해, 자체 제작된 형광상관분광장치로 임의적으로 희석된 시료들의 농도를 약 0.1 nM ~ 10 nM의 범위에서 측정할 수 있었고, 양자점의 확산계수를 $27{\pm}1{\mu}m^2/s$로 결정할 수 있었다.

• Journal of the Optical Society of Korea
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• 제12권3호
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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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• 제18권5호
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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.

• 대한화학회지
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• 제55권5호
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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.

• 대한화학회지
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• 제47권5호
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• pp.447-459
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• 2003

일반화된 이차원 상관 분광학은 모든 분자 분광학 즉, 적외선 (infrared, IR), 근적외선 (near-infrared, NIR), 라만 (Raman) 및 형광 (fluorescence) 분광학뿐만 아니라 X-ray 회절, X-ray 흡수 분광학 (XAS), 크로마토그래피 (chromatography) 등에 적용되어 시간뿐만 아니라 온도, 압력, 농도, 조성과 같은 다른 물리적 변화인 외부 섭동 (perturbation) 아래에서 얻은 스펙트럼 분석의 새로운 분광학으로 다양한 분야의 연구가 활발히 진행 중이다. 또한 같은 외부 섭동 아래에서 얻은 완전히 다른 두 스펙트럼의 2D hetero-spectral correlation analysis가 가능하므로 다양한 분야 즉, 생체 물질, 고분자, 나노입자 등의 연구가 활발히 진행 중이다. 이런 다양한 분야의 응용성 때문에 ？일반？?이차원 상관 분광학？？물리, 분석, 고분자, 나노물질 및 생화학 연구에 새로운 방향을 제시할 수 있을 뿐만 아니라 바이오 나노기술 연구에 상승 효과를 제공할 수 있다. 본 논문은 "일반화된 이차원 상관 분광학"의 원리와 그 다양한 응용성을 본 저자들의 연구를 중심으로 소개하고자 한다.

• Journal of Biosystems Engineering
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• 제35권2호
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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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• 제32권5호
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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.