• Title/Summary/Keyword: Probe preparation

Search Result 90, Processing Time 0.026 seconds

Construction and analysis of painting probe for homogeneously staining regions in human neuroblastoma cell line IMR-32

  • Park, Sun-Hwa;Kim, Ho-Chung;Chun, Yong-Hyuck
    • Journal of Genetic Medicine
    • /
    • v.1 no.1
    • /
    • pp.45-50
    • /
    • 1997
  • Neuroblastoma, a pediatric malignant neoplasm of neural crest origin, has a wide range of clinical virulence. The mechanisms contributing to the development of neuroblastomas are largely unclear, but non-random chromosomal changes identified over the past years suggest the involvement of genetic alterations. Amplification of the human N-myc proto-oncogene is frequently seen either in extrachromosomal double minutes or in homogeneously staining regions (HSRs) of aggressively growing neuroblastomas. N-myc maps to chromosome 2 band 24, but HSR have never been observed at this band, suggesting transposition of N-myc during amplification. We have constructed and analyzed the region-specific painting probe for HSR in neuroblastoma IMR-32 to determine the derivative chromosomes. Microdissection was performed on HSR using an inverted microscope with the help of microglass needles and an micromanipulator. We pretreated the microdissected fragments with Topoisomerase I which catalyzes the relaxation of supercolled DNA, and performed two initial rounds of DNA synthesis with T7 DNA polymerase followed by conventional PCR to enable the reliable preparation of Fluorescent in situ hybridization probe from a single microdissected chromosome. With this method, it was possible to construct the region-specific painting probe for HSR. The probe hybridized specifically to the HSRs of IMR-32, and to 2p24, 2p13 of normal chromosome. Our results suggest there was coamplification of N-myc together with DNA of the chromosome 2p24 and 2p13. Moreover, the fluorescent signals for the amplified chromosomal regions in IMR-32 cells were also easily recognized at a Thus this painting probe can be applied to detect the similar amplification of N-myc in neuroblastoma tissue, and the probe pool for HSR may be used to identify the cancer-relevant genes.

  • PDF

A Brief Overview of Atom Probe Tomography Research

  • Gault, Baptiste
    • Applied Microscopy
    • /
    • v.46 no.3
    • /
    • pp.117-126
    • /
    • 2016
  • Atom probe tomography (APT) has been fast rising in prominence over the past decade as a key tool for nanoscale analytical characterization of a range of materials systems. APT provides three-dimensional mapping of the atom distribution in a small volume of solid material. The technique has evolved, with the incorporation of laser pulsing capabilities, and, combined with progress in specimen preparation, APT is now able to analyse a very range of materials, beyond metals and alloys that used to be its core applications. The present article aims to provide an overview of the technique, providing a brief historical perspective, discussing recent progress leading to the state-of-the-art, some perspectives on its evolution, with targeted examples of applications.

Detection of Plant RNA Viruses by Hybridization Using In Vitro Transcribed RNA Probes (In Viro 전사 RNA Probe를 이용한 식물 바이러스병의 진단)

  • 최장경;이종희;함영일
    • Korean Journal Plant Pathology
    • /
    • v.11 no.4
    • /
    • pp.367-373
    • /
    • 1995
  • The cDNAs derived from the coat protein (CP) genes of six plant RNA viruses, tobacco mosaic virus-pepper strains (TMV-P) and -ordinary strain (TMV-OM), potato virus Y (PVY), turnip mosaic virus (TuMV), cucumber mosaic virus (CMV) and potato leafroll virus (PLRV), were subcloned into the transcription vector, pSPT18, containing SP6 and T7 promoters. The digoxigenin (DIG)-labeled RNA polymerase after linearlization of the cloned pSPTs with XbaI or SacI, and were tested for their sensitivities for the detection of the six viruses. In slot-blot hybridization, dilution end points for the detection of TMV-P and TMV-OM were 10-4, while those of PVY, TuMV and CMV were 10-3. PLRV was detected at the dilution of 10-2. When each RNA probe was applied for the detection of the viruses in the preparations from the leaf disks (8 mm in diameter, and 12 to 15 mg in weight) of infected natural host plants, TMV-P, TMV-OM and TuMV could be detected from one disk, while PVY from 1 or 2 disks. CMV was detected in the preparation from two disks, and PLRV from three disks. With DIG-labeled RNA probe, PVY was detected at 5 days after inoculation, but with ELISA the virus was detected at 8 days after inoculation to tobacco (Nicotiana tabacum cv. Xanthi nc) plants on which symptoms appeared at 9 days after inoculation. No difference was observed in cross reaction between the RNA probes for the detection of TMV-P and TMV-OM.

  • PDF

Preparation and Properties of UV Curable Urethane Acrylates for Ink Binder (Ink Binder용 UV 경화형 조성물의 제조 및 특성)

  • Park, Hyun-Ju;Han, Chang-Duk;Oh, Sang-Taek
    • Journal of Adhesion and Interface
    • /
    • v.14 no.4
    • /
    • pp.167-174
    • /
    • 2013
  • In this study, Ultraviolet (UV) curable urethane acrylates for ink binder was prepared by reaction of IPDI, polyether polyol and HEA. The UV curing behavior of compositions with HEA/PETA ratio on UV irradiation time was investigated by FT-IR spectrophotometer, probe tack tester and Photo-DSC. Conversion % and gel content were increased with increasing UV irradiation time, but probe tack was decreased. Storage modulus, tensile strength and decomposition temperature were increased as PETA content increased. In case of HEA/PETA ratio was 30/70, adhesion property of UV-cured composition on PMMA sheet was excellent.

Array-based Nano-amplification Technique Was Applied in Detection of Hepatitis E Virus

  • Liu, Hui-Hui;Cao, Xuan;Yang, Yong;Liu, Ming-Gui;Wang, Ye-Fu
    • BMB Reports
    • /
    • v.39 no.3
    • /
    • pp.247-252
    • /
    • 2006
  • A rapid method for the detection of Hepatitis E Virus (HEV) was developed by utilizing nano-gold labeled oligonucleotide probes, silver stain enhancement and the microarray technique. The 5'-end -$NH_2$ modified oligonucleotide probes were immobilized on the surface of the chip base as the capture probe. The detection probe was made of the 3'-end -SH modified oligonucleotide probe and nano-gold colloid. The optimal concentrations of these two probes were determined. To test the detection sensitivity and specificity of this technique, a conservative fragment of the virus RNA was amplified by the RT-PCR/PCR one step amplification. The cDNA was hybridized with the capture probes and the detection probes on microarray. The detection signal was amplified by silver stain enhancement and could be identified by naked eyes. 100 fM of amplicon could be detected out on the microarray. As the results, preparation of nano-gold was improved and faster. Development time also was shortened to 2 min. Thus, considering high efficiency, low cost, good specificity and high sensitivity, this technique is alternative for the detection of HEV.

THREE DIMENSIONAL ATOM PROBE STUDY OF NI-BASE ALLOY/LOW ALLOY STEEL DISSIMILAR METAL WELD INTERFACES

  • Choi, Kyoung-Joon;Shin, Sang-Hun;Kim, Jong-Jin;Jung, Ju-Ang;Kim, Ji-Hyun
    • Nuclear Engineering and Technology
    • /
    • v.44 no.6
    • /
    • pp.673-682
    • /
    • 2012
  • Three dimensional atom probe tomography (3D APT) is applied to characterize the dissimilar metal joint which was welded between the Ni-based alloy, Alloy 690 and the low alloy steel, A533 Gr. B, with Alloy 152 filler metal. While there is some difficulty in preparing the specimen for the analysis, the 3D APT has a truly quantitative analytical capability to characterize nanometer scale particles in metallic materials, thus its application to the microstructural analysis in multi-component metallic materials provides critical information on the mechanism of nanoscale microstructural evolution. In this study, the procedure for 3D APT specimen preparation was established, and those for dissimilar metal weld interface were prepared near the fusion boundary by a focused ion beam. The result of the analysis in this study showed the precipitation of chromium carbides near the fusion boundary between A533 Gr. B and Alloy 152.

Early Stages of Preparation for Fatherhood (예비 아버지의 부성경험에 관한 일 연구)

  • Hwang, Jung-hae;Baik, Kyung-im
    • Korean Journal of Child Studies
    • /
    • v.25 no.5
    • /
    • pp.59-71
    • /
    • 2004
  • Based on ethnographic methods, the present research utilized individual interviews of 1 to 2 hours each with various probe questions to discover early stages of preparation for fatherhood. Interviews of the 12 male participants took place in naturalistic settings. Most were between 28 to 32 years of age and middle SES class. Results showed that during their wife's pregnancy expectant fathers come to terms with becoming a father; they obtain a new understanding of life and make efforts to behave desirably in society; attitudes towards their own parents change; they have a different relationship with their wife; relations with close associates change, and occupational identity changes in that feelings of responsibility for a wife and child becomes more sincere.

  • PDF

TEM sample preparation using micro-manipulator for in-situ MEMS experiment

  • Hyunjong Lee;Odongo Francis Ngome Okello;Gi-Yeop Kim;Kyung Song;Si-Young Choi
    • Applied Microscopy
    • /
    • v.51
    • /
    • pp.8.1-8.7
    • /
    • 2021
  • Growing demands for comprehending complicated nano-scale phenomena in atomic resolution has attracted in-situ transmission electron microscopy (TEM) techniques for understanding their dynamics. However, simple to safe TEM sample preparation for in-situ observation has been limited. Here, we suggested the optical microscopy based micro-manipulating system for transferring TEM samples. By adopting our manipulator system, several types of samples from nano-wires to plate-like thin samples were transferred on micro-electro mechanical systems (MEMS) chip in a single step. Furthermore, the control of electrostatic force between the sample and the probe tip is found to be a key role in transferring process.

Effects of Preparation conditions of sintered GdBa$_2$Cu$_3$Ox on the microstructures and electrical properties (소결체 GdBa$_2$Cu$_3$Ox의 제조 조건이 미세구조 및 전기적 특성에 미치는 영향)

  • 김시열;임호빈
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
    • /
    • 1988.10a
    • /
    • pp.102-104
    • /
    • 1988
  • GdBa$_2$Cu$_3$Ox superconductors were prepared by ceramic powder compacting method. X-ray powder diffraction patterns and SEM microphotographs were taken to analyze phase and microstructure, and electrical properties were investigated by 4-point probe method. The results show that the $N_2$atmosphere enhances the formation of high Tc(critical temperature) phase, but the Tc is independent of sintering conditions.

  • PDF

Dimethylnitrosamine-Induced Reduction in the Level of Poly-ADP-Ribosylation of Histone Proteins of Blood Lymphocytes - a Sensitive and Reliable Biomarker for Early Detection of Cancer

  • Kma, Lakhan;Sharan, Rajeshwar Nath
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.15 no.15
    • /
    • pp.6429-6436
    • /
    • 2014
  • Poly-ADP-ribosylation (PAR) is a post-translational modification of mainly chromosomal proteins. It is known to be strongly involved in several molecular events, including nucleosome-remodelling and carcinogenesis. In this investigation, it was attempted to evaluate PAR level as a reliable biomarker for early detection of cancer in blood lymphocyte histones. PAR of isolated histone proteins was monitored in normal and dimethylnitrosamine (DMN)-exposed mice tissues using a novel ELISA-based immuno-probe assay developed in our laboratory. An inverse relationship was found between the level of PAR and period of DMN exposure in various histone proteins of blood lymphocytes and spleen cells. With the increase in the DMN exposure period, there was reduction in the PAR level of individual histones in both cases. It was also observed that the decrease in the level of PAR of histones resulted in progressive relaxation of genomic DNA, perhaps triggering activation of genes that are involved in initiation of transformation. The observed effect of carcinogen on the PAR of blood lymphocyte histones provided us with a handy tool for monitoring biochemical or physiological status of individuals exposed to carcinogens without obtaining biopsies of cancerous tissues, which involves several medical and ethical issues. Obtaining blood from any patient and separating blood lymphocytes are routine medical practices involving virtually no medical intervention, post-procedure medical care or trauma to a patient. Moreover, the immuno-probe assay is very simple, sensitive, reliable and cost-effective. Therefore, combined with the ease of preparation of blood lymphocytes and the simplicity of the technique, immuno-probe assay of PAR has the potential to be applied for mass screening of cancer. It appears to be a promising step in the ultimate goal of making cancer detection simple, sensitive and reliable in the near future.