• Title/Summary/Keyword: Fluorescent dyes

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Moutan Cortex Extract Inhibits Amyloid ${\beta}$ Protein (25-35)-induced Neurotoxicity in Cultured Rat Cortical Neurons (Amyloid ${\beta}$ 2 Protein (25-35) 유도 배양신경세포 독성에 대한 목단피의 억제효과)

  • Kim, Joo-Youn;Ju, Hyun-Soo;Ban, Ju-Yeon;Song, Kyung-Sik;Seong, Yeon-Hee
    • Korean Journal of Medicinal Crop Science
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    • v.16 no.6
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    • pp.409-415
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    • 2008
  • Moutan cortex, the root bark of Paeonia suffruticosa Andrews (Paeoniaceae), has pharmacological effects such as anti-inflammatory, antiallergic, analgesic and antioxidant activities. We investigated a methanol extract of Moutan cortex for neuroprotective effects on neurotoxicity induced by amyloid ${\beta}$ protein ($A{\beta}$) (25-35) in cultured rat cortical neurons. Exposure of cultured cortical neurons to $10\;{\mu}M\;A{\beta}$ (25-35) for 24 h induced neuronal apoptotic death. Moutan cortex inhibited $10\;{\mu}M\;A{\beta}$ (25-35)-induced neuronal cell death at 30 and $50\;{\mu}g/m{\ell}$, which was measured by a 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) assay and Hoechst 33342 staining. Moutan cortex inhibited $10\;{\mu}M\;A{\beta}$ (25-35)-induced elevation of intracellular calcium concentration ($[Ca^{2+}]_i$), and generation of reactive oxygen species (ROS) which were measured by fluorescent dyes. Moutan cortex also inhibited glutamate release into medium induced by $10\;{\mu}M\;A{\beta}$ (25-35), which was measured by HPLC. These results suggest that Moutan cortex prevents $A{\beta}$ (25-35)-induced neuronal cell damage by interfering with the increase of $[Ca^{2+}]_i$, and then inhibiting glutamate release and ROS generation. Moutan cortex may have a therapeutic role in preventing the progression of Alzheimer's disease.

Multimodal Nonlinear Optical Microscopy for Simultaneous 3-D Label-Free and Immunofluorescence Imaging of Biological Samples

  • Park, Joo Hyun;Lee, Eun-Soo;Lee, Jae Yong;Lee, Eun Seong;Lee, Tae Geol;Kim, Se-Hwa;Lee, Sang-Won
    • Journal of the Optical Society of Korea
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    • v.18 no.5
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    • pp.551-557
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    • 2014
  • In this study, we demonstrated multimodal nonlinear optical (NLO) microscopy integrated simultaneously with two-photon excitation fluorescence (TPEF), second-harmonic generation (SHG), and coherent anti-Stokes Raman scattering (CARS) in order to obtain targeted cellular and label-free images in an immunofluorescence assay of the atherosclerotic aorta from apolipoprotein E-deficient mice. The multimodal NLO microscope used two laser systems: picosecond (ps) and femtosecond (fs) pulsed lasers. A pair of ps-pulsed lights served for CARS (817 nm and 1064 nm) and SHG (817 nm) images; light from the fs-pulsed laser with the center wavelength of 720 nm was incident into the sample to obtain autofluorescence and targeted molecular TPEF images for high efficiency of fluorescence intensity without cross-talk. For multicolor-targeted TPEF imaging, we stained smooth-muscle cells and macrophages with fluorescent dyes (Alexa Fluor 350 and Alexa Fluor 594) for an immunofluorescence assay. Each depth-sectioned image consisted of $512{\times}512$ pixels with a field of view of $250{\times}250{\mu}m^2$, a lateral resolution of $0.4{\mu}m$, and an axial resolution of $1.3{\mu}m$. We obtained composite multicolor images with conventional label-free NLO images and targeted TPEF images in atherosclerotic-plaque samples. Multicolor 3-D imaging of atherosclerotic-plaque structural and functional composition will be helpful for understanding the pathogenesis of cardiovascular disease.

Single-Protein Molecular Interactions on Polymer-Modified Glass Substrates for Nanoarray Chip Application Using Dual-Color TIRFM

  • Kim, Dae-Kwang;Lee, Hee-Gu;Jung, Hyung-Il;Kang, Seong-Ho
    • Bulletin of the Korean Chemical Society
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    • v.28 no.5
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    • pp.783-790
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    • 2007
  • The immobilization of proteins and their molecular interactions on various polymer-modified glass substrates [i.e. 3-aminopropyltriethoxysilane (APTS), 3-glycidoxypropyltrimethoxysilane (GPTS), poly (ethylene glycol) diacrylate (PEG-DA), chitosan (CHI), glutaraldehyde (GA), 3-(trichlorosilyl)propyl methacrylate (TPM), 3'-mercaptopropyltrimethoxysilane (MPTMS), glycidyl methacrylate (GMA) and poly-l-lysine (PL).] for potential applications in a nanoarray protein chip at the single-molecule level was evaluated using prismtype dual-color total internal reflection fluorescence microscopy (dual-color TIRFM). A dual-color TIRF microscope, which contained two individual laser beams and a single high-sensitivity camera, was used for the rapid and simultaneous dual-color detection of the interactions and colocalization of different proteins labeled with different fluorescent dyes such as Alexa Fluor® 488, Qdot® 525 and Alexa Fluor® 633. Most of the polymer-modified glass substrates showed good stability and a relative high signal-to-noise (S/N) ratio over a 40-day period after making the substrates. The GPTS/CHI/GA-modified glass substrate showed a 13.5-56.3% higher relative S/N ratio than the other substrates. 1% Top-Block in 10 mM phosphate buffered saline (pH 7.4) showed a 99.2% increase in the blocking effect of non-specific adsorption. These results show that dual-color TIRFM is a powerful methodology for detecting proteins at the single-molecule level with potential applications in nanoarray chips or nano-biosensors.

The Fabrication and Characteristic Analysis of Single-Layer White Organic Light Emitting Devices (단일층 백색유기발광소자의 제작 및 특성분석)

  • Kim, Jung-Yeoun;Kang, Seong-Jong;Roh, Byeong-Gyu;Kang, Myung-Koo;Oh, Hwan-Sool
    • Journal of the Institute of Electronics Engineers of Korea TE
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    • v.39 no.2
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    • pp.11-16
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    • 2002
  • In this paper, single-layer white organic light emitting device was fabricated on ITO glass substrate using PVK as host, Bu-PBD as electron transport layer, Nile Red, Coumarin 6, TPB as red, green, blue color fluorescent dyes. The red, green, blue organic light emitting devices were fabricated respectively. After the characteristic analysis of each color device, the white organic light emitting device was fabricated with optimized condition of each color device by spin coating method. we obtained white emission CIE coordination of (0.32, 0.34) and luminescence of 785cd/$m^2$ at driving voltage of 20V with condition of PVK(70wt%), Bu-PBD(30wt%), Nile Red(0.015mol%), Coumarin 6(0.04mol%), TPB(3mol%). 

Synthesis of New Spiro[benzo[c]fluorene-7,9'-fluorene] Dimers and Their Optical Properties

  • Seo, Jeong-A;Lee, Chil-Won;Gong, Myoung-Seon
    • Bulletin of the Korean Chemical Society
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    • v.34 no.5
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    • pp.1414-1420
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    • 2013
  • Five novel spiro[benzo[c]fluorene-7,9'-fluorene] based dyes, including 5-[spiro[benzo[c]fluorene-7,9'-fluoren]-5-yl] spiro[benzo[c]fluorene-7,9'-fluorene] (7), 5-[spiro[benzo[c]fluorene-7,9'-fluoren]-9-yl] spiro[benzo[c]fluorene-7,9'-fluorene] (8), 5-[spiro[benzo[c]fluorene-7,9'-fluoren]-2'-yl] spiro[benzo[c]fluorene-7,9'-fluorene] (9), 9-[spiro[benzo[c]fluorene-7,9'-fluoren]-9-yl] spiro[benzo[c]fluorene-7,9'-fluorene] (10), and 2'-[spiro[benzo[c]-fluorene-7,9'-fluoren]-2'-yl] spiro[benzo[c]fluorene-7,9'-fluorene] (11) were successfully prepared from the corresponding halogen and boronic acid derivatives through the Suzuki coupling reaction, respectively. Chemical structures were confirmed by $^1H$ nuclear magnetic resonance (NMR), $^{13}C$ NMR, Fourier transforminfrared spectrscopy, mass spectroscopy, and elemental analysis. The thermal properties were determined by differential scanning calorimetry and thermal gravimetric analysis. The relationships between the optical and electrochemical properties and the combined positions between these dimers were systematically investigated using UV-vis, photoluminescence (PL), and photoelectron spectroscopy. These five dimers exhibited high fluorescent quantum yields and good morphological stability with high glass transition states > $174^{\circ}C$. Dimer 7 showed a UV absorbance peak at 353 nm, emission PL peak at 424 nm, and quantum efficiency of 0.62 in a cyclohexane solution.

Comparison of Scattered Light of ex vivo Mouse Neutrophils by Different Wavelength Laser Irradiation (2개의 레이저 파장에 따른 마우스 호중구의 산란광 비교 연구)

  • Park, Jae-Sung;Son, Min-Ji;Hwang, Chang-Soon;Lee, Ho
    • Journal of the Korean Society of Industry Convergence
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    • v.25 no.3
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    • pp.365-378
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    • 2022
  • Complete blood cell count(CBC) is a technique that counts leukocytes for each type of blood cell being analyzed. The principle is that laser is incident to ex vivo flowing leukocytes in a microcapillary tube and scattered light occurs by laser and leukocytes. By collecting the scattered light, we can count the types of cells because different cells generate different light-scattering patterns. However, the technique has an intrinsic limitation, scattering pattern is shown in a wide range region in the resulting, which makes it difficult to accurate analyze and use fluorescent dyes. To overcome this limitation, a new design of CBC with a dual laser, which irradiates with orthogonal angles for collecting quad-scattering information was proposed. Before development, the scattering difference depending on wavelength must be investigated to only catch up to the scattered signal by angles. Some studies, which focused on simple particles, have been conducted to theoretically and experimentally investigate different scatterings by wavelength. In this study, we propose an optical system for measuring scattered light and investigate a complex particle. As a result, the green laser made strong scattering signals in both the forward and side direction: 10% and 30%, respectively.

Healing of the bone around pure titanium implants without primary bone contact (초기 골 접촉이 없는 순수 티타늄 임프란트 주위 골의 치유반응)

  • Ahn, Jae-Hyun;Kim, Heung-Joong;Park, Joo-Cheol;Han, Kyung-Yoon;Kim, Byung-Ock
    • Journal of Periodontal and Implant Science
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    • v.29 no.1
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    • pp.233-249
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    • 1999
  • Primary fixation is one of the most important factor in establishing adequate osseointegration between implant and bone. To evaluate the initial healing response of bone around implants without primary bone contact, this study was designed to create considerable space between implant and bone in 5 mongrel dogs, about 1-year old. After 3 holes of 6.0mm in diameter were prepared at the femur neck of the dogs, commercially pure titanium thread type implants(STERI-$OSS^{(R)}$), 8mm in length and 3.8mm, 5.0mm and 6.0mm in diameter, were inserted. Implants were supported by only nonresorbable membrane($Teflon^{(R)}$), and the penetration of upper soft tissue into the gap was inhibited by it. The each implant was positioned in the center of the drilled hole. 9 implants with different diameters were inserted in 3 dogs for histologic observation, and 12 were inserted in 2 dogs for mobility test and removal torque test.Fluorescent dyes were injected in order of Doxycycline, Alizarin Red S, and Calcein at intervals of 2 weeks. At 4-, 8-, and 12-week after placement, 3 dogs were sacrificed for histologic observation, and at 8- and 12-week after placement, 2 dogs were sacrificed for mobility test using $Periotest^{(R)}$ (Simens AG, Bensheim, Germany) and torque test using Autograph AGS-1000D $series^{(R)}$(Japan). The result were as follows: 1. The wider the gap between bone and implant was, the less bone maturity was, and the later osseointegration was occurred. Trabecular direction of new bone around implant was changed from parallel to perpendicular to the implant, and the gap was filled with new bone, over time. 2. There was a decreasing tendency over time in the mobility of all implants, but the wider gap between bone and implant was, the smaller decrease of the mobility was. 3. There was a increasing tendency over time in the removal torque gauge of all implants, and the wider gap was, the smaller increase of the removal torque gauge was. The results suggest that osseointegration in case of implant without primary bone contact may be obtained by guided bone regeneration technique with prolonged healing period, but the time of second surgery should be considered carefully.

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Healing of the Bone around Hydroxyapatite-Coated Implants without Primary Bone Contact (초기 골 접촉이 없는 수산화 인회석 피복 임프란트 주위 골의 치유)

  • Cho, Hyung-Soo;Shin, Kwang-Yong;Kim, Heung-Joong;Park, Joo-Cheol;Han, Kyung-Yoon;Kim, Byung-Ock
    • Journal of Periodontal and Implant Science
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    • v.29 no.2
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    • pp.415-433
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    • 1999
  • Implant stability is the key to long-term successful outcome for osseointegrated implants. To evaluate the initial healing response of bone around HA-coated implants without primary bone contact. 21 HA-coated thread type implants(STERI-OSS?) were placed in the femurs of 5 mongrel dogs, about 1-year old. Implants, 8 mm in length and 3.8mm(experimental 1group), 5.0mm(experimental 2group) and 6.0mm(control group) in diameter, were inserted after 3 holes of 6.0mm in diameter and 10mm in depth were prepared in the surgical sites each dog. Implants were supported by only nonresorbable membrane($Teflon^{(R)}$), in order to prevent the ingrowth of upper soft tissue into the gap between bone and implant, and to maintain each implant to be positioned in the center of the drilled hole. 9 implants with different diameters were inserted in 3 dogs for histologic observation, and 12 implants were inserted in 2 dogs for mobility test and removal torque test. Fluorescent dyes were injected for the observation of new bone formation in order of $Terramycin^{(R)}$, Arizarin $Red^{(R)}$, and $Calcein^{(R)}$ at an interval of 2 weeks. 3 dogs were sacrificed for histologic observation at 4, 8, and 12-week after placement. Light microscopy and confocal laser scanning microscopy were used to qualitatively characterize the bone around HA-coated implant. 2 dogs were sacrificed for mobility test($Periotest^{(R)}$, Simens AG, Bensheim, Germany) and removal torque test($Autograph^{(R)}$ AGS-1000D series, Japan) at 8 and 12-week after placement The results were as follows: 1. Histologic observation showed that osseointegration occurred to both control and experimental groups as time lapse, but delayed bone healing was revealed in 3.8mm group (experimental 1group), compared to contrtol group and 5.0mm group (experimental 2group). 2. The mobility test showed that the experimental groups had no distinguishable movement during experimental periods of 8 and 12-week, and there was no difference in mobility depending on the gap between bone and implant, and time lapse. 3. The removal torque forces were increased depended on the gaps decreasing between bone and implant, and time lapse. The results suggest that HA-coated implant without primary bone contact, based on guided bone regeneration could obtain its stability in all experimental groups as time lapse, but bone healing was delayed in experimental group of 3.8mm. And the results suggested that studies on correlationship between mobility test and removal torque test for implant stability would be necessary.

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The Physical Characteristics of the flow field and the Form of Arrested Salt Wedge (정상 염수쇄기의 형상과 흐름 장의 물리적 특성)

  • 이문옥
    • 한국해양학회지
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    • v.25 no.2
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    • pp.62-73
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    • 1990
  • An experimental study is performed in order to catch the characteristics of the flow field at arrested salt wedge, using a rectangular open channel. Arrested salt wedge is generally so stable that the observations are easy, but velocities and interfacial waves are measured with the aid of visualization method, by injection of fluorescent dyes. The density interface, which is defined as the zone of maximum density variation with depth, exists in about 0.5 cm below the visual interface, and vertical density profile is quite well satisfied with Homeborn model. Interfacial layer has high turbulent intensity and its thickness decreases as the overall Richardson number increases and has magnitude of roughly 17% of upper layer. Cross-sectional velocity distribution just shows the influence of a side-wall friction and in the upper layer vertical velocity profile also becomes uniformly as Reynolds number increases, but in the lower layer it shows nearly parabolic type. Supposes that we divide salt wedge into three domains, that is, river mouth, intermediate and tip zone, entertainment coefficient is small at the intermediate zone and large at the river mouth and the tip zone. River mouth or intermediate zone has comparatively stable interface and capillary wave therefore s produced and propagated downstream. On the other hand, tip zone is very unstable, cusping ripple or bursting ripple is then produced incessantly. Arrested salt wedge form is nearly linear and has no relation to densimetric Froude number and Reynolds number.

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Development of Molecular Diagnosis Using Multiplex Real-Time PCR and T4 Phage Internal Control to Simultaneously Detect Cryptosporidium parvum, Giardia lamblia, and Cyclospora cayetanensis from Human Stool Samples

  • Shin, Ji-Hun;Lee, Sang-Eun;Kim, Tong Soo;Ma, Da-Won;Cho, Shin-Hyeong;Chai, Jong-Yil;Shin, Eun-Hee
    • Parasites, Hosts and Diseases
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    • v.56 no.5
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    • pp.419-427
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    • 2018
  • This study aimed to develop a new multiplex real-time PCR detection method for 3 species of waterborne protozoan parasites (Cryptosporidium parvum, Giardia lamblia, and Cyclospora cayetanensis) identified as major causes of traveler's diarrhea. Three target genes were specifically and simultaneously detected by the TaqMan probe method for multiple parasitic infection cases, including Cryptosporidium oocyst wall protein for C. parvum, glutamate dehydrogenase for G. lamblia, and internal transcribed spacer 1 for C. cayetanensis. Gene product 21 for bacteriophage T4 was used as an internal control DNA target for monitoring human stool DNA amplification. TaqMan probes were prepared using 4 fluorescent dyes, $FAM^{TM}$, $HEX^{TM}$, $Cy5^{TM}$, and CAL Fluor $Red^{(R)}$ 610 on C. parvum, G. lamblia, C. cayetanensis, and bacteriophage T4, respectively. We developed a novel primer-probe set for each parasite, a primer-probe cocktail (a mixture of primers and probes for the parasites and the internal control) for multiplex real-time PCR analysis, and a protocol for this detection method. Multiplex real-time PCR with the primer-probe cocktail successfully and specifically detected the target genes of C. parvum, G. lamblia, and C. cayetanensis in the mixed spiked human stool sample. The limit of detection for our assay was $2{\times}10$ copies for C. parvum and for C. cayetanensis, while it was $2{\times}10^3$ copies for G. lamblia. We propose that the multiplex real-time PCR detection method developed here is a useful method for simultaneously diagnosing the most common causative protozoa in traveler's diarrhea.