• Title/Summary/Keyword: Fluorescence Microscope

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Pharmacognostic Evaluation of the Flower of Alcea rosea L.

  • Mehrotra, Shanta;Rawat, A.K.S.;Shome, Usha
    • Natural Product Sciences
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    • v.5 no.1
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    • pp.39-47
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    • 1999
  • The flowers of Alcea rosea L., Malvaceae, sold in the Indian market under the trade name 'Gulkhairo', are well known for their expectorant, cooling and diuretic properties and used in many indigenous cough mixtures in India. The present paper deals with the detailed pharmacognosy of the floral parts including morphological, anatomical, phytochemical and fluorescence characters. Some of the diagnostic features of the drug are : pedicel characterized by multicellular appendages, stellate hairs, rosette crystals of Ca-oxalate, starch sheath and large sized mucilage canals; sepals having distinctive multicellular appendages arranged in a semilunar fashion present adaxially at their base; monadelphous stamens, pollen grains pentaporate provided with dimorphic spines; placentation axile, ovules campylotropous; dark green fluorescence of the powder with nitrocellulose in amyl acetate and yellow fluorescence of trichomes under Fluorescence microscope.

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Single-molecule Detection of Fluorescence Resonance Energy Transfer Using Confocal Microscopy

  • Kim, Sung-Hyun;Choi, Don-Seong;Kim, Do-Seok
    • Journal of the Optical Society of Korea
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    • v.12 no.2
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    • pp.107-111
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    • 2008
  • We demonstrated single-molecule fluorescence resonance energy transfer (FRET) from single donor-acceptor dye pair attached to a DNA with a setup based on a confocal microscope. Singlestrand DNAs were immobilized on a glass surface with suitable inter-dye distance. Energy transfer efficiency between the donor and the acceptor dyes attached to the DNA was measured with different lengths of DNA. Photobleaching of single dye molecule was observed and used as a sign of single-molecule detection. We could achieve high enough signal-to-noise ratio to detect the fluorescence from a single-molecule, which allows real-time observation of the distance change between single dye pairs in nanometer scale.

Construction and Patterning of the biospecific affinity surfaces on gold using dendrimer

  • Hong, Mi-Yeong;Yun, Hyeon-Cheol;Kim, Hak-Seong
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.703-706
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    • 2000
  • We constructed a biospecific affinity surface using hyper-branched dendrimers on gold for biospecific recognition, and characterized the resulting surfaces by using confocal fluorescence microscopy. The dendrimer monolayer was firstly constructed on the mercaptoundecanoic acid SAM/Au with pentafluorophenyl ester activation and further functionalized with sulfo-NHS-biotin, an activated ester of biotin. To confirm the formation of biospecific affinity surface, FITC(fluorescein isothiocyanate)-labeled avidin was loaded onto the biotinylated dendrimer monolayer, and fluorescence images of the bound avidins were investigated with a confocal microscope. The constructed biospecific affinity surface showed a much more dense and uniform fluorescence compared to those from poly-L-lysine- and cystamine SAM-based affinity surfaces. For the dependency on the concentration of added FITC-labeled avidin on the affinity surface, derived fluorescence could be detectable from as low as $1{\mu}g/ml$, and intensified up to $50{\mu}g/ml$. Further reaction of FITC-labeled avidin layer with TMR(tetramethylrhodamine)-biocytins resulted in the efficient FRET(fluorescence resonance energy transfer) phenomenon. As an extension of the study, we attempted a patterning of the affinity surfaces on gold by microcontact printing. Fluorescence of the patterned surface demonstrated that FITC-labeled avidin molecules were specifically bound to the biotinylated patches.

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Study of a Brain Tumor and Blood Vessel Detection System Using Multiple Fluorescence Imaging by a Surgical Microscope (수술현미경에서의 다중형광영상을 이용한 뇌종양과 혈관영상 검출 시스템 연구)

  • Lee, Hyun Min;Kim, Hong Rae;Yoon, Woong Bae;Kim, Young Jae;Kim, Kwang Gi;Kim, Seok Ki;Yoo, Heon;Lee, Seung Hoon;Shin, Min Sun;Kwon, Ki Chul
    • Korean Journal of Optics and Photonics
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    • v.26 no.1
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    • pp.23-29
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    • 2015
  • In this paper, we propose a microscope system for detecting both a tumor and blood vessels in brain tumor surgery as fluorescence images by using multiple light sources and a beam-splitter module. The proposed method displays fluorescent images of the tumor and blood vessels on the same display device and also provides accurate information about them to the operator. To acquire a fluorescence image, we utilized 5-ALA (5-aminolevulinic acid) for the tumor and ICG (Indocyanine green) for blood vessels, and we used a beam-splitter module combined with a microscope for simultaneous detection of both. The beam-splitter module showed the best performance at 600 nm for 5-ALA and above 800 nm for ICG. The beam-splitter is flexible to enable diverse objective setups and designed to mount a filter easily, so beam-splitter and filter can be changed as needed, and other fluorescent dyes besides 5-ALA and ICG are available. The fluorescent images of the tumor and the blood vessels can be displayed on the same monitor through the beam-splitter module with a CCD camera. For ICG, a CCD that can detect the near-infrared region is needed. This system provides the acquired fluorescent image to an operator in real time, matching it to the original image through a similarity transform.

Microscope Examination of Attached Biofilm under Anaerobic Conditions (혐기성 조건에서 담체에 부착된 미생물의 관찰)

  • 박성열;김도한;나영수;박영식;송승구
    • Journal of Environmental Health Sciences
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    • v.27 no.1
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    • pp.100-105
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    • 2001
  • Microstructural examinations were performed on the anaerobic biofilm from reactor filled with PE support media. Optical microscope, SEM and fluorescent microscope were used for qualitative and morphological studies on the attached microorganism under anaerobic condition. Microorganisms were attached in crevices where protection from shear forces of surfaces where easy to contact with support media surface. A hypothesis for biofilm accumulation occurs on a surface such as polymer support media is presented schematically : 1st step ; cell-support media attachment, 2nd step ; cell-support media attachment and cell-cell attachment, 3rd step ; attached biofilm from neighboring crevices joins together and growing, 4th step ; mature and irregualar biofilm was formed. In SEM photographs, shape and structures of biofilm were observed, but microorganism species and methanogens were not identified. A large number of methanogenic bacteria were identified on the surface of PE substratum by fluorescence under 480nm of radiation and it was estimated that methanogenic bacteria was related to initial attachment of bacteria under anaerobic condition.

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Development of An Integration Management System of Analyzing Fluorescence Images on Smart Phone (모바일용 형광이미지 분석 통합관리 시스템 개발)

  • Cho, Mi-Gyung;Shim, Jae-Sool
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2012.05a
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    • pp.916-919
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    • 2012
  • Bioimaging that can be imaging phenomena within cells of a molecular size have been advanced in technology. We can observe clearly DNA and proteins using a confocal microscope. Currently biological fluorescent imaging area is used essentially for diagnosis and treatment in health and clinical care field. In this paper, we developed an integration management system of analyzing fluorescence images on smart phone. It can support a user to analyse fluorescence images anytime anywhere. And our system is based on client-server configuration and has functions that can figure intensity of fluorescence images and manage many imaging data. Proposed system can be a mean of ubiquitous health because it helps a doctor diagnose by analyzing fluorescence images of emergency patients without time and space restrictions.

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Development of Image-based System for Multiple Fluorescence Imaging Study (다중형광영상 연구를 위한 영상기반 시스템 개발)

  • Yoon, WoongBae;Kim, Hong Rae;Lee, Hyun Min;Kim, Young Jae;Kim, Kwang Gi;Yoo, Heon;Lee, Seung Hoon
    • Journal of Korea Multimedia Society
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    • v.18 no.12
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    • pp.1445-1452
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    • 2015
  • In these days, fluorescent materials such as ICG or 5-ALA is used for the brain surgery. The patients who underwent brain tumor surgery has been increased during last 30 years and the survivorship rate increased 22∼33% in 5 years. Recently, the Fluorescence induction surgery is developed for more safety and improved the resection rate for the glioma in the neurosurgery field. In this study, we proposed fluorescence area detection method for ICG and 5-ALA fluorescence induced surgery using acquired images from image processing. Accuracy was 99.21% from ICG images, and 99.51% from 5-ALA images. Matthews correlation coefficient was 88.67% from ICG images, and 90.49% from 5-ALA images.

Development of line-scanning two-photon microscopy based on spatial and temporal focusing for tryptophan based auto fluorescence imaging (고속 트립토판 자가형광 이미징을 위한 시공간적 집중 기반의 라인 스캐닝 이광자 현미경 개발)

  • Lee, Jun Ho;Nam, Hyo Seok;Kim, Ki Hean
    • Journal of the Korean Society of Visualization
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    • v.11 no.2
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    • pp.41-45
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    • 2013
  • Two-photon microscopy (TPM) is minimally-invasive 3D fluorescence microscopy based on nonlinear excitation, and TPM can visualize cellular structures based on auto-fluorescence. Line-scanning TPM is one of high-speed TPM methods without sacrificing the image resolution by using spatial and temporal focusing. In this paper, we developed line-scanning TPM based on spatial and temporal focusing for auto-fluorescence imaging by exciting the tryptophan. Laser source for this system was an optical parametric oscillator (OPO) and it made near 570 nm femtosecond pulse laser. It had 200fs pulse width and 1.72 nm bandwidth, so that the achievable depth resolution was 2.41um and field of view (FOV) is 10.8um. From the characterization, our system has 3.0 um depth resolution and 12.3 um FOV. We visualized fixed leukocyte cell sample and compared with point scanning system.

DETECTION OF OCCLUSAL CARIES USING LASER FLUORESCENCE (레이저 형광법의 교합면 우식증 탐지 효과)

  • Kim, Chang-Gi;Lee, Chang-Seop;Lee, Sang-Ho
    • Journal of the korean academy of Pediatric Dentistry
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    • v.29 no.4
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    • pp.600-606
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    • 2002
  • The purpose of this study was to evaluate the diagnostic validity of an incipient occlusal caries using argon laser fluorescence. Extracted human premolars and molars with enamel carious lesion of occlusal surface were assessed using visual examination, visual examination with probing, argon laser fluorescence and histologic depth of carious lesion. The results in each of all the three detection methods were compared to the assessment of histologic depth of carious lesion using polarized microscope. The results from the present study can be summarized as follows; 1. There was highly correlation between the histologic depth of occlusal caries and all three detection methods(P<0.01). 2. The reproducibility(kappa value) of the visual examination, visual examination with probing and argon laser fluorescence between the histologic depth of occlusal caries was 0.189, 0.128, 0.472. The highest correlation was seen between detection of occlusal caries by argon laser fluorescence and histologic scores by polarized microscope. The results from this study indicated that argon laser fluorescence considered to be accurate and reliable method in detecting occlusal caries.

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VIP IMMUNOREACTIVITY IN THE RAT TRIGEMINAL GANGLION AFTER INFERIOR ALVEOLAR NERVE AXOTOMY (하치조신경 절단 후 흰쥐 삼차신경절에서 VIP-IR의 변화)

  • Kim, Heung-Joong;Park, Joo-Cheol;Kim, Hyun-Sub;Moon, Joo-Hoon
    • Restorative Dentistry and Endodontics
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    • v.25 no.2
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    • pp.225-234
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    • 2000
  • The purpose of this study was to investigate the distribution and fluorescene intensity of vasoactive intestinal polypeptide(VIP) immunoreactive cells in rat trigeminal ganglion after inferior alveolar nerve axotomy. The animals were divided into normal and two experimental groups. The experimental animals were sacrificed at 14th and 28th day after inferior alveolar nerve axotomy. The trigeminal ganglion was removed and immersed in the 4% paraformaldehyde-0.2% picric acid in 0.1M phosphate buffer. Serial frozon sections about $16{\mu}m$ in thickness were cut with a cryostat. The immunofluorescence staining was performed. The rabbit anti-VIP(1 : 8,000) was used as primary antibody and fluorescene isothiocynate(FITC)-conjugated anti-rabbit IgG(1 : 80) as secondary antibody. The slides were observed under confocal laser scanning microscope. Three-dimensional images were constructed from 9 serial images(each $1{\mu}m$ in thickness) made by automatic optical sectioning. Unprocessed optical sections were obtained and stored on a optical disk. Color picture were printed by a video copy processor. The results were as follows; 1. The appearance of VIP immunoreactive cells in the mandibular part of trigeminal ganglion was 8.79${\pm}$1.99% in normal group and 39.16${\pm}$5.62% in 14 days, 16.25${\pm}$2.39% in 28 days after inferior alveolar nerve axotomy groups. 2. The relative fluorescence intensity of VIP immunoreactive cell bodies in the mandibular part of trigeminal ganglion was 134.40${\pm}$10.39 in normal group and 192.88${\pm}$14.06 in 14 days, 143.10${\pm}$5.02 in 28 days after nerve axotomy groups. Therefore, the relative fluorescence intensity of 14 days after nerve axotomy group was 43.3% higher than intensity of normal group. 3. In optical single section analysis of VIP immunoreactive cell bodies, white cell bodies(moderate fluorescence intensity) were the most abundant in normal and 28 days after nerve axotomy groups. Whereas, in 14 days after nerve axotomy group, red cell bodies(high fluorescence intensity) were the most abundant. 4. In optical serial section analysis of VIP immunoreactive cell bodies, red cell bodies(high fluorescence intensity) were observed in a part of the 9 sections of normal and 24 days after nerve axotomy groups. Whereas, red cell bodies were observed in all of the 9 sections of 14 days after nerve axotomy group. 5. The results indicates that number and fluorescence intensity of VIP immunoreactive cells were increased in the mandibular part of trigeminal ganglion following inferior alveolar nerve axotomy.

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