• Title/Summary/Keyword: Fluorescence probe technique

Search Result 68, Processing Time 0.022 seconds

Characterization of the Surface Contribution to Fluorescence Correlation Spectroscopy Measurements

  • Chowdhury, Salina A.;Lim, Man-Ho
    • Bulletin of the Korean Chemical Society
    • /
    • v.32 no.2
    • /
    • pp.583-589
    • /
    • 2011
  • Fluorescence correlation spectroscopy (FCS) is a sophisticated and an accurate analytical technique used to study the diffusion of molecules in a solution at the single-molecule level. FCS is strongly affected by many factors such as the stability of the excitation power, photochemical processes, mismatch between the refractive indices, and variations in the cover glass thickness. We have studied FCS near the surface of a cover glass by using rhodamine 123 as a fluorescent probe and have observed that the surface has a strong influence on the measurements. The temporal autocorrelation of FCS decays with two characteristic times when the confocal detection volume is positioned near the surface of the cover glass. As the position of the detection volume is moved away from the surface, the FCS autocorrelation becomes one-component decaying; the characteristic time of the decay is the same as the faster-decaying component in the FCS autocorrelation near the surface. This observation suggests that the faster component can be attributed to the free diffusion of the probe molecules in the solution, while the slow component has its origin from the interaction between the probe molecules and the surface. We have characterized the surface contribution to the FCS measurements near the surface by changing the position of the detection volume relative to the surface. The influence of the surface on the diffusion of the probe molecules was monitored by changing the chemical properties of the surface. The surface contribution to the temporal autocorrelation of the FCS strongly depends on the chemical nature of the surface. The hydrophobicity of the surface is a major factor determining the surface influence on the free diffusion of the probe molecules near the surface.

Sex Determination of In Vitro Fertilized Bovine Embryos by Fluorescence in situ Hybridization Technique

  • Han, M. S.;E. J. Cho;H. B. Ha;Park, H. S.;S. H. Sohn
    • Proceedings of the KSAR Conference
    • /
    • 2004.06a
    • /
    • pp.287-287
    • /
    • 2004
  • Sexing from bovine embryos which were fertilized in vitro implicate a possibility of production of the sex controlled cattle. This study was carried out to investigate the possibility of determining of embryo sex by fluorescence in situ hybridization (FISH) technique. FISH was achieved in in vitro fertilized bovine embryos using a bovine Y-specific DNA probe which constructed from the btDYZ-1 sequences. (omitted)

  • PDF

Identification of a Human Y Chromosome Specific DNA Probe and Their Clinical Application by Fluorescence in situ Hybridization Techniques (사람 Y 염색체 특이 DNA Probe의 개발과 이를 이용한 FISH 기술의 임상적 적용)

  • Ryu, Ran-Suk;Lee, Sang-Chan;Lee, Chae-Sik;Kim, Jong-Heung;Ryu, Eun-Koung;Sohn, Sea-Hwan
    • Clinical and Experimental Reproductive Medicine
    • /
    • v.27 no.4
    • /
    • pp.405-415
    • /
    • 2000
  • Objective: The present study was undertaken to synthesize a human Y chromosome specific probe and to confirm the usefulness of the probe for fluorescence in situ hybridization (FISH) in various types of human cells. Methods: An approximately 400 bp DNA fragment of the DYZ1 sequences was synthesized by PCR using digoxigenin labeled dUTP (dig-PCR). The fidelity of probe was tested by FISH for cultured and uncultured human lymphocytes, amniocytes, chorionic villus cells, embryos, sperms, and germ cells of seminiferous tubule. Results: The human Y chromosome specific probe hybridized specifically to Y chromosome of the cells that had been tested. This probe assigned to the Yq12 region where the DYZ1 repetitive sequence is concentrated. Conclusion: We have identified a human Y chromosome specific probe that hybridized specifically to the Y chromosome by FISH for various types of uncultured as well as cultured cells. Therefore FISH technique using human Y chromosome specific probe should be useful for clinical application as a diagnostic tool for the detection of human Y chromosome.

  • PDF

Cure Monitoring of Am Epoxy-Anhydride System by Means of Fluorescence Spectroscopy (형광분석기를 이용한 에폭시-산무수물계의 경화 모니터링)

  • 조동환;김득수;이종근
    • Polymer(Korea)
    • /
    • v.25 no.2
    • /
    • pp.199-207
    • /
    • 2001
  • In the present study the cure behavior of diglycidyl ether of bisphenol-A(DGEBA) using an anhydride-based hardener in the presence of N,N-dimethyl benzyl amine (BDMA) or 1-cyanoethyl-2-ethyl-4-methyl imidazole (2E4MZ-CN) as an accelerator has been monitored and interpreted from the viewpoint of photophysical properties by means of fluorescence spectroscopy. To do this, 1,3-bis-(1-pyrene)propane (BPP) was well incorporated in the epoxy resin system by mechanical blending. The BPP probe, which is very sensitive to conformational change of the molecule influenced by the surrounding medium, successfully formed intramolecular excimer fluorescence. It is susceptible to the micro-viscosity or local viscosity and molecular mobility according to the epoxy cure. The cure behavior was explained with monomer fluorescence intensity ($I_{M}$ ), excimer fluorescence intensity ($I_{E}$ ) and $I_{M}$ /$I_{E}$ ratio as a function of cure time, cure temperature and accelerator. The present work agreed with the previous report on the cure behavior of an epoxy-anhydride system studied using DSC or torsion pendulum method. This study also suggests that the use of fluorescence technique may provide information on cure behavior of a thermosetting resin in a low temperature region, which has not been well interpreted by other analytical methods.

  • PDF

The Region of Distribution of Barbiturates in Synaptosomal Plasma Membrane Vesicles Isolated from Rat Brain as Studied by Fluorescence Quenching (Barbiturates가 생체세포막 외측 단층의 소수성 부위와 친수성 부위에 분포되는 상대적 비율)

  • Yun, Il;Lee, Byung-Woo
    • The Korean Journal of Pharmacology
    • /
    • v.31 no.3
    • /
    • pp.271-278
    • /
    • 1995
  • The relative distribution ratio of barbiturates between hyarocarbon interior and surface region of outer monolayer of synaptosomal plasma membrane vesicles (RSPMV) isolated from rat whole brain was determined by employing the fluorescent probe technique. The two fluorescent probes N- octadecylnaphthyl-2-amine-6-sulfonic acid (ONS) and 12-(9-anthroyloxy) stearic acid (AS) were utilized as probes for hydrocarbon interior and surface of outer monolayer of RSPMV. respectively. The Stern-Volmer equation for fluorescent quenching was modified to calculate the relative distribution ratio. The analysis of preferential quenching of these probes by barbiturates indicates that pentobarbital, hexobarbital, amobarbital and phenobarbital are predominantly distributed on the surface region. whereas thiopental sodium has an accessibility to the hydrocarbon interior of the outer monolayer of the RSPMV. From these results, it is strongly suggested that the more effective penetration into the hydrocarbon interior of the outer monolayer of the membrane lipid bilayer could result in higher general anesthetic activity.

  • PDF

Effects of Parathyroid Hormone on the Fluidity of the Plasma Membrane Vesicles of Cultured Osteoblasts

  • Kang, Jung-Sook
    • Journal of Photoscience
    • /
    • v.8 no.3_4
    • /
    • pp.87-92
    • /
    • 2001
  • Intramolecular excimer formation of 1,3-di(1-pyrenyl)propane (Py-3-Py) and fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene (DPH) were used to investigate the effects of parathyroid hormone (PTH) on the bulk bilayer fluidity of the plasma membrane vesicles isolated from cultured osteoblasts (OB-PMV). In a dose-dependent manner, rat PTH-(1-34) [rPTH-(1-34)] increased the excimer to monomer fluorescence intensity ratio (I'/I) of Py-3-Py and decreased the anisotropy (r) of DPH in OB-PMV. This indicates that PTH increased both the lateral and rotational diffusion of the probes in OB-PMY. Selective quenching of DPH fluorescence by trinitrophenyl groups was utilized to examine the transbilayer fluidity asymmetry of OB-PMV. The anisotropy, limiting anisotropy, and order parameter of DPH in the inner monolayer were 0.024, 0.032, and 0.062 greater than calculated for the outer monolayer of OB-PMY. Selective quenching of DPH fluorescence by trinitrophenyl groups was also utilized to examine the transbilayer effects of PTH on the fluidity of OB-PMV. rPTH-(1-34) had a greater fluidizing effect on the outer monolayer as compared to the inner monolayer of OB-PMV. Thus, it has been proven that PTH exhibits a selective rather than nonselective fluidizing effect within transbilayer domains of OB-PMV.

  • PDF

Synthesis and characterization of ABA types tri-block copolymers derived from p-dioxanone, ${\varepsilon}-caprolactone$ and poly(ethylene glycol)

  • Remant Bahadur K.C.;Bhattarai Shanta Raj;Aryal Santosh;Khil, Myung-Seob;Kim, Hak-Yong;Lee, Douk-Rae
    • Proceedings of the Polymer Society of Korea Conference
    • /
    • 2006.10a
    • /
    • pp.255-255
    • /
    • 2006
  • Polymeric nanoparticles are recognized as promising drug carriers [1]. Here, novel tri-block copolymers based on poly PPDO, PCL and PEG were synthesized and employed for the formulation of reproducible polymeric nanoparticles [2]. To estimate the feasibility of the polymer to form polymeric nanoparticles, nanoparticles were prepared by co-solvent evaporation technique. Polymerization and structural features of the polymer were analyzed by different physico-chemical techniques. Existence of hydrophobic domains as a core of nanoparticles was characterized by $^{1}H-NMR$ spectroscopy, and further confirmed by fluorescence technique using pyrene as probe.

  • PDF

Developing a Cantilever-type Near-field Scanning Optical Microscope Using a Single Laser for Topography Detection and Sample Excitation

  • Ng'ang'a, Douglas Kagoiya;Ali, Luqman;Lee, Yong Joong;Byeon, Clare Chisu
    • Current Optics and Photonics
    • /
    • v.5 no.3
    • /
    • pp.229-237
    • /
    • 2021
  • The capabilities of the near-field scanning optical microscope (NSOM) for obtaining high resolution lateral topographical images as well as for mapping the spectroscopic and optical properties of a sample below the diffraction limit of light have made it an attractive research field for most researchers dealing with optical characteristics of materials in nano scales. The apertured NSOM technique involves confining light into an aperture of sub-wavelength size and using it to illuminate a sample maintained at a distance equal to a fraction of the sub-wavelength aperture (near-field region). In this article, we present a setup for developing NSOM using a cantilever with a sub-wavelength aperture at the tip. A single laser is used for both cantilever deflection measurement and near-field sample excitation. The laser beam is focused at the apex of the cantilever where a portion of the beam is reflected and the other portion goes through the aperture and causes local near-field optical excitation of the sample, which is then raster scanned in the near-field region. The reflected beam is used for an optical beam deflection technique that yields topographical images by controlling the probe-sample in nano-distance. The fluorescence emissions signal is detected in far-field by the help of a silicon avalanche photodiode. The images obtained using this method show a good correlation between the topographical image and the mapping of the fluorescence emissions.

Telomere의 양적 분석을 이용한 닭의 bio-marker개발

  • 조은정;최철환;전익수;박철;손시환
    • Proceedings of the Korea Society of Poultry Science Conference
    • /
    • 2004.11a
    • /
    • pp.13-15
    • /
    • 2004
  • Telomeres are the end of chromosomes and consist of a tandem repeat sequence of (TTAGGG)n and associated proteins. Telomeres are essential for chromosome stability and are related with cell senescence and apoptosis. This study was carried out to analyze the amount of telomeric DNA of chicken lymphocytes, which is to considered as bio-marker. The amount of telomeric DNA of lymphocytes in Korean Native Chicken and White Leghorn was analyzed by quantitative-fluorescence in situ hybridization (Q-FISH) technique using the chicken telomeric DNA probe. Telomere quantifies were compared among breeds, ages and sex, and the relationship between the amount of telomeres and their productive trait was also analyzed. Comparing the amount of telomeric DNA on lymphocytes during growing period, the amount of telomeres was gradually decreased as growing older. The telomere quantity was also significantly different in breeds and sex. Estimating correlation coefficient, the amount of telomeres was positively correlated to sexual maturity and body weight but negatively correlated to hen day egg production and egg weight. These results implicate the telomere quantity is considered as an individual bio-marker.

  • PDF

Membrane-Ordering Effects of Barbiturates on Pure Phospholipid Model Membranes

  • Knag, Jung-Sook;Chung, Young-Za;Cho, Goon-Jae;Byun, Won-Tan;Yun, Il
    • Archives of Pharmacal Research
    • /
    • v.15 no.3
    • /
    • pp.196-203
    • /
    • 1992
  • Intramolecular excimer formation of 1, 3-di(1-pyrenyl)propane (Py-3-Py) and fluorescence polarization of 1, 6-diphenyl-1, 3, 5-hexatriene (DPH) were used to investigate the effects of barbiturates on the fluidity of model membranes of phosphatidycholine (SPMVPC), phosphatidylserine (SPMVPS), and phosphatidylinositol (SPMVPI) fractions of synaptosomal plasma membrane vesicles (SPMV) isolated from bovine cerebral cortex. In a dose-dependent manner, barbiturates decreased the excimer to monomer fluorescence intensity ratio (I'/I) of Py-3-Py and increased the anisotropy(r), rotational relaxation time (P), limiting anisotropy $(r_infty)$, and order parameter (S) of DPH in SPMVPC, SPMVPS and SPMVPI. This indicates that barbiturates decreased both the lateral and rotational diffusion of the probes in SPMVPC, SPMVPS and SPMVPI. The relative potencies of barbiturates in ordering the membranes were in the order: pentobarbital > hexobarbital > amobarbital > phenobarbital. This order correlates well with the anesthetic potencies of barbiturates and the potencies for enhancement of $\gamma$-aminobutyric acid-stimulated chloride uptake. Thus, it is strongly suggested that a close relationship might exist between the membrane ordering effects of barbiturates and the chloride fluxes across SPMV.

  • PDF