• Title/Summary/Keyword: Fluorescence labeling

Search Result 52, Processing Time 0.024 seconds

Fluorescence Detection of Cell Death in Liver of Mice Treated with Thioacetamide

  • Kang, Jin Seok
    • Toxicological Research
    • /
    • v.34 no.1
    • /
    • pp.1-6
    • /
    • 2018
  • The purpose of this study was to detect cell death in the liver of mice treated with thioacetamide (TAA) using fluorescence bioimaging and compare this outcome with that using conventional histopathological examination. At 6 weeks of age, 24 mice were randomly divided into three groups: group 1 (G1), control group; group 2 (G2), fluorescence probe control group; group 3 (G3), TAA-treated group. G3 mice were treated with TAA. Twenty-two hours after TAA treatment, G2 and G3 mice were treated with Annexin-Vivo 750. Fluorescence in vivo bioimaging was performed by fluorescence molecular tomography at two hours after Annexin-Vivo 750 treatment, and fluorescence ex vivo bioimaging of the liver was performed. Liver damage was validated by histopathological examination. In vivo bioimaging showed that the fluorescence intensity was increased in the right upper part of G3 mice compared with that in G2 mice, whereas G1 mice showed no signal. Additionally ex vivo bioimaging showed that the fluorescence intensity was significantly increased in the livers of G3 mice compared with those in G1 or G2 mice (p < 0.05). Histopathological examination of the liver showed no cell death in G1 and G2 mice. However, in G3 mice, there was destruction of hepatocytes and increased cell death. Terminal deoxynucleotidyl transferase dUTP nick end labeling staining confirmed many cell death features in the liver of G3 mice, whereas no pathological findings were observed in the liver of G1 and G2 mice. Taken together, fluorescence bioimaging in this study showed the detection of cell death and made it possible to quantify the level of cell death in male mice. The outcome was correlated with conventional biomedical examination. As it was difficult to differentiate histological location by fluorescent bioimaging, it is necessary to develop specific fluorescent dyes for monitoring hepatic disease progression and to exploit new bioimaging techniques without dye-labeling.

Methods of measuring presynaptic function with fluorescence probes

  • Yeseul Jang;Sung Rae Kim;Sung Hoon Lee
    • Applied Microscopy
    • /
    • v.51
    • /
    • pp.2.1-2.7
    • /
    • 2021
  • Synaptic vesicles, which are endogenous to neurotransmitters, are involved in exocytosis by active potentials and release neurotransmitters. Synaptic vesicles used in neurotransmitter release are reused via endocytosis to maintain a pool of synaptic vesicles. Synaptic vesicles show different types of exo- and endocytosis depending on animal species, type of nerve cell, and electrical activity. To accurately understand the dynamics of synaptic vesicles, direct observation of synaptic vesicles is required; however, it was difficult to observe synaptic vesicles of size 40-50 nm in living neurons. The exo-and endocytosis of synaptic vesicles was confirmed by labeling the vesicles with a fluorescent agent and measuring the changes in fluorescence intensity. To date, various methods of labeling synaptic vesicles have been proposed, and each method has its own characteristics, strength, and drawbacks. In this study, we introduce methods that can measure presynaptic activity and describe the characteristics of each technique.

Labeling of oral spirochetes with fluorescent fatty acids

  • Hong, Jin;Kim, Kyu-Joong;Lee, Si-Young
    • International Journal of Oral Biology
    • /
    • v.33 no.2
    • /
    • pp.65-70
    • /
    • 2008
  • Oral spirochetes are anaerobes known as one of causative agents for periodontal diseases. In this study, we investigated the possibility of utilizing fluorescent fatty acids for labeling oral spirochetes. Bacterial labeling was standardized with three different lengths of fluorescent fatty acids: 5-octadecanoylaminfluorescein (OAF), 5-dodecanoylamin-fluorescein (DAF), and 5-hexadecanoylaminfluorescein (HAF). Among these fatty acids, OAF showed the best labeling activity. Treponema denticola ATCC 35405 was totally saturated to the maximum when incubated with OAF $1\;{\mu}g/ml$ for 1 hour. Treponema vincentii LA-1 also increased in fluorescence in proportion to incubation time length and the concentration. In conclusion, these findings showed the possibility that the fluorescent fatty acid can be used for labeling oral spirochetes.

Toxicity and Biomedical Imaging of Fluorescence-Conjugated Nanoparticles in Hematopoietic Progenitor Cells

  • Min, Gye-Sik;Kim, Dong-Ku
    • Reproductive and Developmental Biology
    • /
    • v.35 no.4
    • /
    • pp.503-510
    • /
    • 2011
  • Cellular uptake of nanoparticles for stem cell labeling and tracking is a critical technique for biomedical therapeutic applications. However, current techniques suffer from low intracellular labeling efficiency and cytotoxic effects, which has led to great interest in the development of a new labeling strategy. Using silica-coated nanoparticles conjugated with rhodamine B isothiocyanate (RITC) (SR), we tested the cellular uptake efficiency, biocompatibility, proliferation or differentiation ability with murine bone marrow derived hematopoietic stem/progenitor cells. The bone marrow hematopoietic cells showed efficient uptake with SR with dose or time dependent manner and also provided a higher uptake on hematopoietic stem/progenitor cells. Biocompatibility tests revealed that the SR had no deleterious effects on cell cytotoxicity, proliferation, or multi-differentiation capacities in vitro and in vivo. SR nanoparticles are advantageous over traditional labeling techniques as they possess a high level of cellular internalization without limiting the biofunctionality of the cells. Therefore, SR provides a useful alternative for gene or drug delivery into hematopoietic stem/progenitor cells for basic research and clinical applications.

Fluorescence-labelling for analysis of protein in starch using asymmetrical flow field-flow fractionation (AF4)

  • Yoo, Yeongsuk;Choi, Jaeyeong;Zielke, Claudia;Nilsson, Lars;Lee, Seungho
    • Analytical Science and Technology
    • /
    • v.30 no.1
    • /
    • pp.1-9
    • /
    • 2017
  • Starch is a mixture of amylose (AMY) and amylopectin (AMP) which are different in physical properties such as molar mass (M), rms radius ($R_g$) and hydrodynamic diameter ($d_H$). The rheological and functional properties of starch are influenced by various factors including the molecular size, molar mass distribution (MD) and the concentration ratio of AMY and AMP. It is also important to analyze proteinaceous material in starch as they affect the flavor and texture of food to which starch is added. In this study, asymmetrical flow field-flow fractionation (AF4) was employed for separation and quantitation of AMY and AMP in starches (Amaranth, potato, taros and quinoa). AF4 was coupled with a multi-angle light scattering (MALS) and a refractive index (RI) detector for determination of the absolute M, MD and molecular structure. It was found that AMP has the M and $R_g$ ranging $3.7{\times}10^7{\sim}6.5{\times}10^8g/mol$ and 84 ~ 250 nm, respectively. Also the existence of branch was confirmed in higher M. In addition, proteinaceous material in starch was analyzed by AF4 coupled with a fluorescence detector (FS) after fluorescence-labeling. AF4-FS with fluorescence-labelling showed a potential for investigation on existence of proteinaceous material and the interaction between proteinaceous material and polysaccharide in starch.

FISH기법 적용을 위한 Y 염색체 특이 DNA Probe의 개발

  • 조은정;류란숙;류은경;손시환
    • Proceedings of the KSAR Conference
    • /
    • 2003.06a
    • /
    • pp.24-24
    • /
    • 2003
  • Fluorescence in situ Hybridization(FISH)는 특정 염기서열을 이용하여 염색체나 염색체상의 DNA위치를 확인하는 기술로서, 면역세포화학 기술과 결합되어져 현미경으로 이들의 유전적 활성도를 직접 확인할 수 있는 방법으로 지금까지의 radioisotopes 대신 non-radioactive labeling 방법으로서 fluorescence을 이용한 분자세포유전학적 검정 방법이다. 따라서 특정 염색체의 FISH probe의 개발은 FISH 기법을 이용하여 조직 또는 세포내 특정 염색체나 DNA의 존재나 이상 유무를 신속하고 정확하게 파악할 수 있다. 본 연구는 소와 사람을 대상으로 Y-염색체 특이 DNA probe를 개발하고 이를 이용하여 FISH를 시행함으로서 본 probe의 신뢰성을 확인하고 임상적 적용 가능성을 제시 하고자 하였다.

  • PDF

Albumin-conjugated Cadmium Sulfide Nanoparticles and their Interaction with KB Cells

  • Selim, K.M. Kamruzzaman;Kang, Inn-Kyu;Guo, Haiqing
    • Macromolecular Research
    • /
    • v.17 no.6
    • /
    • pp.403-410
    • /
    • 2009
  • Cytotoxicity is a severe problem of cadmium sulfide nanoparticles(CSNPs) for use in biological systems. In the present study, mercaptoacetic acid-coated CSNPs were conjugated with bovine serum albumin (BSA) to improve biocompatibility. The surface properties of the CSNPs and albumin-conjugated CSNPs (ACSNPs) were characterized by XRD, UV, FTIR, EA, TEM and DLS. Human breast cancer cells (KB cells) were then cultured in the presence of the nanoparticles to evaluate the cytotoxicity of CSNPs and ACSNPs. Finally, the fluorescence intensity of the nanoparticles' aqueous solution was examined using a fluorescence spectrometer. The results showed that the cell compatibility and fluorescence intensity of ACSNPs were higher than those of CSNPs. The strongly luminescent features of the biocompatible ACSNPs are promising for use in biological fields such as cellular labeling, intracellular tracking and molecular imaging.

Characterization of Binding of Treponema denticola to Immobilized Fibrinogen using the Fluorescent Fatty Acid Labeling Method

  • Hong, Jin;Lee, Si-Young
    • International Journal of Oral Biology
    • /
    • v.35 no.3
    • /
    • pp.107-111
    • /
    • 2010
  • Treponema denticola is a gram-negative anaerobe that can cause periodontal disease. The adhesion of this bacterium to host tissues is considered to be the primary event in the colonization and infection of a host. Fibrinogen is generally found in damaged tissues resulting from periodontitis. The binding ability of T. denticola to fibrinogen may therefore be an important virulence factor in inducing periodontal diseases. It has been reported recently that oral spirochetes can be labeled with fluorescent fatty acids and we speculated that this labeling method could be used in an oral spirochete binding assay. The binding of several different strains of T. denticola to immobilized human fibrinogen was therefore tested using the fluorescent fatty acid labeling method. In the case of immobilized fibrinogen, the T. denticola ATCC 35405 strain showed saturable binding to immobilized fibrinogen. Indeed, all four different T. denticola strains tested in this experiment, T. denticola ATCC 35405, T. denticola ATCC 33520, T. denticola ATCC 35404 and T. denticola OTK showed binding to fibrinogen. The fluorescent fatty acid labeling method thus shows utility in binding assays for T. denticola, different strains of which can generally bind to immobilized fibrinogen.

INFLUENCE OF A FUNCTIONAL LOADING TIME ON BONE FORMATION AROUND OSSEOINTEGRATED TITANIUM IMPLANTS IN ADULT DOGS (성견에서 골유착성 타이타늄 임프란트의 기능적 노출 시기가 주위의 골형성에 미치는 영향)

  • Yang Ja-Ho;Lee Ho-Yong
    • The Journal of Korean Academy of Prosthodontics
    • /
    • v.29 no.3
    • /
    • pp.55-74
    • /
    • 1991
  • The purpose of this study was to investigate the influence of early functional load around osseointegrated titanium implants. 24 titanium plasma spray coated implants (ITI HS-type) were placed into the previously extracted site in the mandible of six adult dogs. The implants were divided into three groups : the control group was the implants without abutment during the experimental period; the experimental group I was loaded by connecting the contoured abutment after 6 weeks of healing; the experimental group II was loaded after 12 weeks of healing: and the mandibular second premolar and surrounding tissues were selected for natural tooth group to compare the implanted group. All dogs were injected intravenously tetracycline, alizarin red S, and calcein for bone labeling. After the experimental period of 18 weeks, the dogs were sacrificed and longitudinal sections of the bone-implant interface were cut and observed using light microscope, scanning electron microscope, and fluorescence microscope. The results of the study were as follows: 1. Light and scanning electron microscopically, all implant surfaces were well contact with bone tissue at the cortical layer, but some areas of cancellous bone were not contact directly. 2. Fluorescence microscopically, number and size of the new secondary osteons around the implant were increased than those of the natural tooth. 3. Fluorescence microscopically, linear and concentrical fluorescence was observed at or near the surface of all implants, and the bone formation and remodeling of the implants loaded after 6 week of healing were great, and unloaded implants were worst. 4. Fluorescence microscopically, endosteal bone formation was greater than periosteal bone formation at or near the implants. 5. Fluorescence microscopically, number and size of linear and concentric fluorescence was increased at the lingual side than the buccal side of the loaded implants. The result of the study indicate the possibility of the early load to the implant via a prosthesis.

  • PDF