• 제목/요약/키워드: Fluorescence Imaging

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Current Statues of Phenomics and its Application for Crop Improvement: Imaging Systems for High-throughput Screening (작물육종 효율 극대화를 위한 피노믹스(phenomics) 연구동향: 화상기술을 이용한 식물 표현형 분석을 중심으로)

  • Lee, Seong-Kon;Kwon, Tack-Ryoun;Suh, Eun-Jung;Bae, Shin-Chul
    • Korean Journal of Breeding Science
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    • v.43 no.4
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    • pp.233-240
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    • 2011
  • Food security has been a main global issue due to climate changes and growing world population expected to 9 billion by 2050. While biodiversity is becoming more highlight, breeders are confronting shortage of various genetic materials needed for new variety to tackle food shortage challenge. Though biotechnology is still under debate on potential risk to human and environment, it is considered as one of alternative tools to address food supply issue for its potential to create a number of variations in genetic resource. The new technology, phenomics, is developing to improve efficiency of crop improvement. Phenomics is concerned with the measurement of phenomes which are the physical, morphological, physiological and/or biochemical traits of organisms as they change in response to genetic mutation and environmental influences. It can be served to provide better understanding of phenotypes at whole plant. For last decades, high-throughput screening (HTS) systems have been developed to measure phenomes, rapidly and quantitatively. Imaging technology such as thermal and chlorophyll fluorescence imaging systems is an area of HTS which has been used in agriculture. In this article, we review the current statues of high-throughput screening system in phenomics and its application for crop improvement.

$In$ $vitro$ MRI and Characterization of Rat Mesenchymal Stem Cells Transduced with Ferritin as MR Reporter Gene (페리틴 리포터 유전자를 발현하는 백서 중간엽 줄기세포의 특성과 자기공명영상 연구)

  • Shin, Cheong-Il;Lee, Whal;Woo, Ji-Su;Park, Eun-Ah;Kim, Pan-Ki;Song, Hyun-Bok;Kim, Hoe-Suk
    • Investigative Magnetic Resonance Imaging
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    • v.16 no.1
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    • pp.47-54
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    • 2012
  • Purpose : This study was performed to evaluate the characteristics of rat mesenchymal stem cells (RMSCs) transduced with human ferritin gene and investigate $in$ $vitro$ MRI detectability of ferritin-transduced RMSCs. Materials and Methods: The RMSCs expressing both myc-tagged human ferritin heavy chain subunit (myc-FTH) and green fluorescence protein (GFP) were transduced with lentiviurs. Transduced cells were sorted by GFP expression using a fluorescence-activated cell sorter. Myc-FTH and GFP expression in transduced cells were detected by immunofluorescence staining. The cell proliferative ability and viability were assessed by MTT assay. The RMSC surface markers (CD29+/CD45-) were analyzed by flow cytometry. The intracellular iron amount was measured spectrophotometically and the presence of ferritin-iron accumulation was detected by Prussian blue staining. $In$ $vitro$ magnetic resonance imaging (MRI) study of cell phantoms was done on 9.4 T MR scanner to evaluate the feasibility of imaging the ferritin-transduced RMSCs. Results: The myc-FTH and GFP genes were stably transduced into RMSCs. No significant differences were observed in terms of biologic properties in transduced RMSCs compared with non-transduced RMSCs. Ferritin-transduced RMSCs exhibited increased iron accumulation ability and showed significantly lower $T_2$ relaxation time than non-transduced RMSCs. Conclusion: Ferritin gene as MR reporter gene could be used for non-invasive tracking and visualization of therapeutic mesenchymal stem cells by MRI.

Pancreatic Compression during Lymph Node Dissection in Laparoscopic Gastrectomy: Possible Cause of Pancreatic Leakage

  • Ida, Satoshi;Hiki, Naoki;Ishizawa, Takeaki;Kuriki, Yugo;Kamiya, Mako;Urano, Yasuteru;Nakamura, Takuro;Tsuda, Yasuo;Kano, Yosuke;Kumagai, Koshi;Nunobe, Souya;Ohashi, Manabu;Sano, Takeshi
    • Journal of Gastric Cancer
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    • v.18 no.2
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    • pp.134-141
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    • 2018
  • Purpose: Postoperative pancreatic fistula is a serious and fatal complication of gastrectomy for gastric cancer. Blunt trauma to the parenchyma of the pancreas can result from an assistant's forceps compressing and retracting the pancreas, which in turn may result in pancreatic juice leakage. However, no published studies have focused on blunt trauma to the pancreas during laparoscopic surgery. Our aim was to investigate the relationship between compression of the pancreas and pancreatic juice leakage in a swine model. Materials and Methods: Three female pigs were used in this study. The pancreas was gently compressed dorsally for 15 minutes laparoscopically with gauze grasped with forceps. Pancreatic juice leakage was visualized by fluorescence imaging after topical administration of chymotrypsin-activatable fluorophore in real time. Amylase concentrations in ascites collected at specified times was measured. In addition, pancreatic tissue was fixed with formalin, and the histology of the compressed sites was evaluated. Results: Fluorescence imaging enabled visualization of pancreatic juice leaking into ascites around the pancreas. Median concentrations of pancreatic amylase in ascites increased from 46 U/L preoperatively to 12,509 U/L 4 hours after compression. Histological examination of tissues obtained 4 hours after compression revealed necrotic pancreatic acinar cells extending from the surface to deep within the pancreas and infiltration of inflammatory cells. Conclusions: Pancreatic compression by the assistant's forceps can contribute to pancreatic juice leakage. These findings will help to improve the procedure for lymph node dissection around the pancreas during laparoscopic gastrectomy.

Evaluation of Near-infrared Fluorescence-conjugated Peptides for Visualization of Human Epidermal Receptor 2-overexpressed Gastric Cancer

  • Jeong, Kyoungyun;Kong, Seong-Ho;Bae, Seong-Woo;Park, Cho Rong;Berlth, Felix;Shin, Jae Hwan;Lee, Yun-Sang;Youn, Hyewon;Koo, Eunhee;Suh, Yun-Suhk;Park, Do Joong;Lee, Hyuk-Joon;Yang, Han-Kwang
    • Journal of Gastric Cancer
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    • v.21 no.2
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    • pp.191-202
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    • 2021
  • Purpose: A near-infrared (NIR) fluorescence imaging is a promising tool for cancer-specific image guided surgery. Human epidermal receptor 2 (HER2) is one of the candidate markers for gastric cancer. In this study, we aimed to synthesize HER2-specific NIR fluorescence probes and evaluate their applicability in cancer-specific image-guided surgeries using an animal model. Materials and Methods: An NIR dye emitting light at 800 nm (IRDye800CW; Li-COR) was conjugated to trastuzumab and an HER2-specific affibody using a click mechanism. HER2 affinity was assessed using surface plasmon resonance. Gastric cancer cell lines (NCI-N87 and SNU-601) were subcutaneously implanted into female BALB/c nu (6-8 weeks old) mice. After intravenous injection of the probes, biodistribution and fluorescence signal intensity were measured using Lumina II (Perkin Elmer) and a laparoscopic NIR camera (InTheSmart). Results: Trastuzumab-IRDye800CW exhibited high affinity for HER2 (KD=2.093(3) pM). Fluorescence signals in the liver and spleen were the highest at 24 hours post injection, while the signal in HER2-positive tumor cells increased until 72 hours, as assessed using the Lumina II system. The signal corresponding to the tumor was visually identified and clearly differentiated from the liver after 72 hours using a laparoscopic NIR camera. Affibody-IRDye800CW also exhibited high affinity for HER2 (KD=4.71 nM); however, the signal was not identified in the tumor, probably owing to rapid renal clearance. Conclusions: Trastuzumab-IRDye800CW may be used as a potential NIR probe that can be injected 2-3 days before surgery to obtain high HER2-specific signal and contrast. Affibody-based NIR probes may require modifications to enhance mobilization to the tumor site.

Layered Double Hydroxide Nanoparticles for Bio-Imaging Applications (LDH 나노입자 기반의 바이오 이미징 소재)

  • Jin, Wenji;Ha, Seongjin;Lee, Dongki;Park, Dae-Hwan
    • Korean Chemical Engineering Research
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    • v.57 no.4
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    • pp.445-454
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    • 2019
  • Layered double hydroxides (LDHs) nanoparticles have emerged as novel nanomaterials for bio-imaging applications due to its unique layered structure, physicochemical properties, and good biocompatibility. Bio-imaging is one of the most important fields for medical applications in clinical diagnostics and therapeutics of various diseases. Enhanced diagnostic techniques are needed to realize new paradigm for next-generation personalized medicine through nanoscale materials. When nanotechnology is introduced into bio-imaging system, nanoparticle probes can endow imaging techniques with enhanced ability to obtain information about biological system at the molecular level. In this review, we summarize structural features of LDH nanoparticles with current issues of bio-imaging system. LDH nanoparticle probes are also discussed through in vitro as well as in vivo studies in various bio-imaging techniques including fluorescence imaging, magnetic resonance imaging (MRI), positron emission tomography (PET), and computed X-ray tomography (CT), which will have the potential in the development of the advanced nanoparticles with high sensitivity and selectivity.

Antifungal and Plant Growth Promotion Activities of Recombinant Defensin Proteins from the Seed of Korean Radish (Raphanus sativus L.)

  • Hwang, Cher-Won
    • Korean Journal of Environmental Agriculture
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    • v.28 no.4
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    • pp.435-441
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    • 2009
  • In the present study, we analyzed the defensin protein deduced from Korean radish (Raphanus sativus L.) seeds.To express the genes in E. coli, we constructed a recombinant expression vector with a defensin gene, named rKRs-AFP gene isolated from Korean radish seeds. Over expressed rKRs-AFP proteins was separated by SDS-PAGE to determine the purity, and protein concentration was determined by the Bradford method. Antifungal activity was assessed by disk assay method against the tested fungi. As a result, when 500 mL of cell culture were disrupted by sonicator, 32.5 mg total proteins were obtained. The purified protein showed a single band on SDS-PAGE with estimated molecular weight about 6 KDa, consistent with the molecular mass calculated from the deduced amino acid sequence. The purified rKRs-AFP protein showed remarkable antifungal activities against several fungi including Aspergillus niger, Botrytis cinerea causing the gray mold disease, and Candida albicans. In field tests using the purified rKRs-AFP protein, the protein showed the reducing activity of disease spot and the mitigating effect of spreading of disease like agrichemicals. The immuno-assay of rKRs-AFP protein showed that the purified protein entirely accumulated at B. cinerea cytoplasm through the hyphal septa shown by fluorescence imaging. There was no fluorescence inside the cell, when the hypha was incubated without the protein. These all results indicate that the recombinant rKRs-AFP proteins can be utilized as a potential antifungal drug to control harmful plant fungal pathogens.

Non-blinking dendritic crystals from C-dot solution

  • Mewada, Ashmi;Vishwakarma, Ritesh;Patil, Bhushan;Phadke, Chinmay;Kalita, Golap;Sharon, Maheshwar;Sharon, Madhuri
    • Carbon letters
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    • v.16 no.3
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    • pp.211-214
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    • 2015
  • Bio-imaging and drug carriers for delivery have created a huge demand for crystals. Crystals are fascinating materials that have been grown for a long time but obtaining biocompatible fluorescent crystals is a challenging task. We report on the growth of fluorescent crystals using a carbon dot (C-dot) solution by a hydrothermal process. The crystallization pattern of these C-dots exhibited a unique dendritic structure having a feather-like morphology. The growth temperature and pressure were maintained at 60℃ and 200 mmHg, respectively, for crystal growth. A green fluorescence (under UV light) that was observed in the C-dot solution was retained in the crystals formed from the solution. Cytotoxicity studies on Vero cells revealed the crystals to be extremely biocompatible. These fluorescent crystals are extremely well suited for biomedical and optoelectronic applications.

3D Light-Sheet Fluorescence Microscopy of Cranial Neurons and Vasculature during Zebrafish Embryogenesis

  • Park, Ok Kyu;Kwak, Jina;Jung, Yoo Jung;Kim, Young Ho;Hong, Hyun-Seok;Hwang, Byung Joon;Kwon, Seung-Hae;Kee, Yun
    • Molecules and Cells
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    • v.38 no.11
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    • pp.975-981
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    • 2015
  • Precise 3D spatial mapping of cells and their connections within living tissues is required to fully understand developmental processes and neural activities. Zebrafish embryos are relatively small and optically transparent, making them the vertebrate model of choice for live in vivo imaging. However, embryonic brains cannot be imaged in their entirety by confocal or two-photon microscopy due to limitations in optical range and scanning speed. Here, we use light-sheet fluorescence microscopy to overcome these limitations and image the entire head of live transgenic zebrafish embryos. We simultaneously imaged cranial neurons and blood vessels during embryogenesis, generating comprehensive 3D maps that provide insight into the coordinated morphogenesis of the nervous system and vasculature during early development. In addition, blood cells circulating through the entire head, vagal and cardiac vasculature were also visualized at high resolution in a 3D movie. These data provide the foundation for the construction of a complete 4D atlas of zebrafish embryogenesis and neural activity.

Effect of Campsis grandiflora on Antioxidative Activity in UVB-irradiated Human Dermal Fibroblasts (사람 섬유아세포에서 UVB 조사에 대한 능소화 추출물의 항산화 효과)

  • Kim, Jin-Hwa;Lee, Bum-Chun;Zhang, Yong-He;Pyo, Hyeong-Bae
    • YAKHAK HOEJI
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    • v.49 no.2
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    • pp.174-179
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    • 2005
  • The human skin is constantly exposed to environmental irritants such as ultraviolet, smoke, chemicals. Free radicals and reactive oxygen species (ROS) caused by these environmen tal facts play critical roles in cellular damage. These irritants are in themselves damaging to the skin structure but they also participate the immensely complex inflammatory reaction. The purpose of this study was to investigate the skin cell protective effect of Campsis grandiflora extract on the UVB-irradiated human dermal fibroblasts (HDFs). We tested free radical and superoxide scavenging effect in vitro. C. grandiflora extracts had potent radical scavenging effect by 82% at $100{\mu}g/ml$, respectively. For testing intracellular ROS scavenging activity the cultured HDFs were analyzed by increase in DCF fluorescence upon exposure to UVB 20 $MJ/cm^2$ after treatment of C.grandiflora extracts. The results showed that oxidation of CM-DCFDA was inhibited by C.grandiflora extracts effectively and C.grandiflora extracts has a potent free radical scavenging activity in UVB- irradiated HDFs. In ROS imaging using confocal microscope we visualized DCF fluorescence in HDFs directly. In conclusion, our results suggest that C.grandiflora can be effectively used for the prevention of UV-induced adverse skin reactions such as radical production, and skin cell damage.

Growth and Physiological Responses of Quercus acutissima Seedling under Drought Stress

  • Lim, Hyemin;Kang, Jun Won;Lee, Solji;Lee, Hyunseok;Lee, Wi Young
    • Plant Breeding and Biotechnology
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    • v.5 no.4
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    • pp.363-370
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    • 2017
  • In this study, Quercus acutissima seedlings were subjected to drought for 30 days then analyzed to determine their response to water deficit. The growth phenotype, chlorophyll fluorescence response, fresh weight, dry weight, photosynthetic pigment levels, soluble sugar content, and malondialdehyde (MDA) were measured to evaluate the effects of drought on plant growth and physiology. The growth phenotype was observed by infrared (IR) digital thermal imaging after 30 days of drought treatment. The maximum, average, and minimum temperatures of drought-treated plant leaves were $1-2^{\circ}C$ higher than those of the control. In contrast, the fresh and dry weights of the dehydrated leaves were generally lower than those of the control. There were no significant differences between treatments in terms of chlorophyll a, chlorophyll b, total chlorophyll, and carotenoid levels. Nevertheless, for the drought treatment, the $F_v/F_m$ and $F_v/F_o$ ratios (chlorophyll fluorescence response) were lower than those for the control. Therefore, photosynthetic activity was lower in the dehydrated plants than the control. The drought-stressed Q. acutissima S0536 had lower soluble sugar (glucose and fructose) and higher MDA levels than the controls. These findings may explain the early growth and physiological responses of Q. acutissima to dehydration and facilitate the selection of drought-resistant tree families.