• Journal of Microbiology and Biotechnology
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• v.31 no.2
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• pp.233-239
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• 2021

Cyanobacteriochromes (CBCRs) are phytochrome-related photoreceptor proteins in cyanobacteria and cover a wide spectral range from ultraviolet to far-red. A single GAF domain that they contain can bind bilin(s) autocatalytically via heterologous recombination and then fluoresce, with potential applications as biomarkers and biosensors. Here, we report that a novel red/green CBCR GAF domain, SPI1085g2 from Spirulina subsalsa, covalently binds both phycocyanobilin (PCB) and phycoerythrobilin (PEB). The PCB-binding GAF domain exhibited canonical red/green photoconversion with weak fluorescence emission. However, the PEB-binding GAF domain, SPI1085g2-PEB, exhibited an intense orange fluorescence (λabs.max = 520 nm, λfluor.max = 555 nm), with a fluorescence quantum yield close to 1.0. The fluorescence of SPI1085g2-PEB was selectively and instantaneously quenched by copper ions in a concentration-dependent manner and exhibited reversibility upon treatment with the metal chelator EDTA. This study identified a novel PEB-binding cyanobacteriochrome-based fluorescent protein with the highest quantum yield reported to date and suggests its potential as a biosensor for the rapid detection of copper ions.

• Medical Lasers
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• v.10 no.3
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• pp.123-131
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• 2021

Optical imaging modalities with properties of real-time, non-invasive, in vivo, and high resolution for image-guided surgery have been widely studied. In this review, we introduce two optical imaging systems, that could be the core of image-guided surgery and introduce the system configuration, implementation, and operation methods. First, we introduce the optical coherence tomography (OCT) system implemented by our research group. This system is implemented based on a swept-source, and the system has an axial resolution of 11 ㎛ and a lateral resolution of 22 ㎛. Second, we introduce a fluorescence imaging system. The fluorescence imaging system was implemented based on the absorption and fluorescence wavelength of indocyanine green (ICG), with a light-emitting diode (LED) light source. To confirm the performance of the two imaging systems, human malignant melanoma cells were injected into BALB/c nude mice to create a xenograft model and using this, OCT images of cancer and pathological slide images were compared. In addition, in a mouse model, an intravenous injection of indocyanine green was used with a fluorescence imaging system to detect real-time images moving along blood vessels and to detect sentinel lymph nodes, which could be very important for cancer staging. Finally, polarization-sensitive OCT to find the boundaries of cancer in real-time and real-time image-guided surgery using a developed contrast agent and fluorescence imaging system were introduced.

• Journal of Veterinary Clinics
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• v.39 no.6
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• pp.395-399
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• 2022

A 6-year-old spayed, female golden retriever dog was presented with a skin mass on the dorsal region of the right carpus. The cytology result of the region revealed characteristics of mast cell tumors (MCTs). Short wave-infrared fluorescence-guided surgery using Indocyanine green (ICG) was performed to determine the surgical margin of the tumor. ICG was injected intravenously 24 hours before the surgery and the patient was hospitalized and carefully monitored. During the surgery, ICG fluorescence-based surgery was performed to identify the tumor and the surgical margin. The tumor was visible, and the skin mass was resected using NIR device for the guidance of the surgical margin of the tumor. Once the resection was complete, the surgical site was again inspected with SWIR fluorescence imaging to identify residual tumor cells. The resected tumor, using ICG navigation, was classified as low-grade cutaneous MCT and the margin was complete on the histopathological result. We report herein a case of resection of a cutaneous MCT in a dog using SWIR fluorescence imaging ICG which can be potentially used for the identification of tumors and evaluation of the surgical margin for complete resection.

• Journal of the Korean Institute of Electrical and Electronic Material Engineers
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• v.35 no.2
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• pp.149-154
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• 2022

In this paper, laser-induced fluorescence properties of four plastics were characterized through spectrometer analysis for real-time microplastic counting. Recently, environmental problems related to microplastics have emerged. In order to detect microplastics, analysis methods such as FT-IR and Raman are used. However, they have the disadvantages of being time-consuming and requiring a pretreatment process. In most plastic products on the market, 10% to 30% of plasticizers and reinforcing agents are added. Therefore, most microplastics present in seawater and freshwater emit fluorescence signals by 270 nm UV light source regardless of their type due to their molecular structure due to additives. Real-time microplastics counting is possible more easily by using the proposed laser-induced fluorescence detection method because of the fluorescence expression characteristic of 340 nm that appears due to the plasticizer of plastics.

• Journal of Microbiology and Biotechnology
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• v.33 no.12
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• pp.1681-1691
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• 2023

Flavin mononucleotide-binding proteins or domains emit cyan-green fluorescence under aerobic and anaerobic conditions, but relatively low fluorescence and less thermostability limit their application as reporters. In this work, we incorporated the codon-optimized fluorescent protein from Chlamydomonas reinhardtii with two different linkers independently into the redox-responsive split intein construct, overexpressed the precursors in hyperoxic Escherichia coli SHuffle T7 strain, and cyclized the target proteins in vitro in the presence of the reducing agent. Compared with the purified linear protein, the cyclic protein with the short linker displayed enhanced fluorescence. In contrast, cyclized protein with incorporation of the long linker including the myc-tag and human rhinovirus 3C protease cleavable sequence emitted slightly increased fluorescence compared with the protein linearized with the protease cleavage. The cyclic protein with the short linker also exhibited increased thermal stability and exopeptidase resistance. Moreover, induction of the target proteins in an oxygen-deficient culture rendered fluorescent E. coli BL21 (DE3) cells brighter than those overexpressing the linear construct. Thus, the cyclic reporter can hopefully be used in certain thermophilic anaerobes.

• Analytical Science and Technology
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• v.27 no.4
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• pp.187-195
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• 2014

It is very important to minimize the damage of latent fingerprints at enhancing the contrast. This study proves the enhancement effects of cyanoacrylate-fumed latent fingerprints using p-dimethylaminobenzealdehyde (DMAB) on semi-porous surfaces and the influence factors. The latent fingerprints in experiment were developed for cyanoacrylate treatment in a vacuum chamber and used after drying at room temperature for 24 hours. For fluorescence staining, the cyanoacrylate-developed marks using DMAB were sublimated during 48 hours under the different conditions of surface area, temperature, atmospheric pressure. First experiment showed how surface area effects on the sublimation rate and fluorescence intensity by DMAB of particle size and container size. In addition, the fluorescence staining using DMAB with solvent-free contact method had the greatest fluorescence intensity after 36 hours and a low fluorescence intensity over a certain size of surface area. Second experiment showed that the evaporation of DMAB solid crystals got a satisfying result in a temperature of $20^{\circ}C$ and reduced time to get the greatest fluorescence intensity. It took a long time to get a optimum level of fluorescence intensity at $30^{\circ}C$ or more and it was less effective in fluorescence intensity. Third experiment on the pressure indicated that the fluorescence intensity of vacuum was weaker than nonvacuum but it was inapplicable to very high variations in pressure.

• Journal of Microbiology and Biotechnology
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• v.17 no.12
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• pp.1922-1929
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• 2007

A simple whole-cell-based sensing system is proposed for determining the cell mass of H. pluvialis using ultraviolet fluorescence spectroscopy. An emission signal at 368 nm was used to detect the various kinds of green, green-brown, brown-red, and red H. pluvialis cells. The fluorescence emission intensities of the cells were highest at 368 nm with an excitation wavelength of 227 nm. An excitation wavelength of 227 nm was then selected for cell-mass sensing, as the emission fluorescence intensities of the cell suspensions were highest at this wavelength after subtracting the background interference. The emission fluorescence intensities of HPLC-grade water, filtered water, and HPLC-grade water containing a modified Bold's basal medium (MBBM) were measured and the difference was less than 1.6 for the selected wavelengths. Moreover, there was no difference in the emission intensity at 368 nm among suspensions of the various morphological states of the cells. A calibration curve of the fluorescence emission intensities. and cell mass was obtained with a high correlation ($R^2=0.9938$) for the various morphological forms of H. pluvialis. Accordingly, the proposed method showed no significant dependency on the various morphological cell forms, making it applicable for cell-mass measurement. A high correlation was found between the fluorescence emission intensities and the dry cell weight with a mixture of green, green-brown, brown-red, and red cells. In conclusion, the proposed model can be directly used for cell-mass sensing without any pretreatment and has potential use as a noninvasive method for the online determination of algal biomass.

• Proceedings of the KSMRM Conference
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• 2004.09a
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• pp.23-32
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• 2004

The chemotherapy sensitive Lewis lung carcinoma (LLC) and chemotherapy resistant Lewis lung carcinoma (CR-LLC) tumors concurrently implanted in mice, and compare these findings with histological macroscopic observations against 3D reconstruction of Fluorescence Molecular Tomography (FMT) preformed in vivo on the same animals. For the 3D image reconstruction we used 32 laser source images, a flat image and 3D surface rendering that confused for 3D Fluorescence Molecular Imaging (FMI). A minimum of ten tissue sections were analyzed per tumor for quantification of the TUNEL-positive cells, cell-associated Cy5.5-Annexin and vessel-associated Alexa Fluor-Lectin. These are useful apoptosis and angiogenesis markers, and they serve as validation experiments to data obtained in vivousing a Cy5.5-Annexin V conjugate injected intravenously in chemotherapy-treated animals carrying the tumors studied histologically. We detected higher levels of apoptosis and corresponding higher levels of Cy5.5 fluorescence in the LLC vs. the CR-LLC tumors according to tissue depth and these findings confirm that in vivo staining with the Cy5.5-Annexing conjugate correlates well with in vitro TUNEL staining and is consistent with the higher apoptotic index expected from the LLC line. There appeared to be 1.38% more apoptosis for LLC than CR-LLC. Consequently there is good correlation between the histology results and in vivo fluorescence-mediated optical imaging. In conclusion the apoptotic images of 3D FMI were validated by microscopic histological image analysis. This is a significant result for the continuous progress of fluorescence 3D imaging research.

• Journal of Environmental Science International
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• v.22 no.11
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• pp.1403-1413
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• 2013

The effects of cadmium ions ($Cd^{2+}$) on the Chl a fluorescence of Ricciocarpos natans were investigated in order to determine whether Chl fluorescence can be used as a biomarker to estimate the physiological responses of plants to cadmium stress. In all plants treated with $Cd^{2+}$, the image of Fv/Fm, which represents the maximum photochemical efficiency of PSII, changed as the $Cd^{2+}$ concentration increased, when treated for 48 h or more. Changes of ${\Phi}_{PSII}$ and $Q_P$ images were recognized even at 10 ${\mu}M$ $Cd^{2+}$. The Chl a O-J-I-P fluorescence transient was also affected even at 10 ${\mu}M$ $Cd^{2+}$. The fluorescence yield decreased considerably in steps J, I and P in plants treated with $Cd^{2+}$, although a typical polyphasic rise was observed in non-treated plants. The Chl fluorescence parameters, Fm, Fv/Fo, Sm, SFIabs, PIabs and ETo/CS, decreased as the $Cd^{2+}$ concentration increased, while the Mo and Kn parameters increased. Peroxidase activity decreased significantly and catalase activity increased as the $Cd^{2+}$ concentration increased. Because of its sensitivity to $Cd^{2+}$ Ricciocarpos natans is useful in experiments investigating the responses of plants to cadmium exposure. Several parameters (Fm, Fv/Fo, Sm, SFIabs, PIabs, ETo/CS, Mo and Kn) can be applied to determine quantitatively the physiological states of plants under cadmium stress.

• 한국신재생에너지학회:학술대회논문집
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• 2010.06a
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• pp.88.2-88.2
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• 2010

Dye-sensitized solar cells (DSSCs) have attracted considerable attention on account of their high solar energy-to-conversion efficiencies and low cost processes compared to conventional p-n junction solar cells. The mechanism of DSSC is based on the injection of electrons from the photo excited dyes into the conduction band of the semiconductor electrode. The oxidized dye is reduced by the hole injection into either the hole conductor or the electrolyte. Thus, the light harvesting effect of dye plays an important role in capturing the photons and generating the electron/hole pair, as well as transferring them to the interface of the semiconductor and the electrolyte, respectively. We used the organic fluorescence materials which can absorb short wavelength light and emit longer wavelength region where dye sensitize effectively. In this work, the DSSCs were fabricated with fluorescence materials added $TiO_2$ photo-electrode which were sensitized with metal-free organic dyes. The photovoltaic performances of fluorescence aided DSSCs were compared, and the recombination dark current curves and the incident photon-to-current (IPCE) efficiencies were measured in order to characterize the effects of the additional light harvesting effect in DSSC. Electro-optical measurements were also used to optimize the fluorescence material contents on TiO2 photo-electrode surface for higher conversion efficiency (${\eta}$), fill factor (FF), open-circuit voltage (VOC) and short-circuit current (ISC). The enhanced light harvesting effect by the judicious choice/design of the fluorescence materials and sensitizing dyes permits the enhancement of photovoltaic performance of DSSC.