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Highly Multiplexed Fluorescence <i>in Situ</i> Hybridization for <i>in Situ</i> Genomics
Onozato, Maristela L.,Yapp, Clarence,Richardson, Douglas,Sundaresan, Tilak,Chahal, Varun,Lee, Jesse,Sullivan, James P.,Madden, Marisa W.,Shim, Hyo S.,Liebers, Matthew,Ho, Quan,Maheswaran, Shyamala,Hab American Society for Investigative Pathology 2019 The Journal of Molecular Diagnostics Vol. No.
<P>The quantification of changes in gene copy number is critical to our understanding of tumor biology and for the clinical management of cancer patients. DNA fluorescence <I>in situ</I> hybridization is the gold standard method to detect copy number alterations, but it is limited by the number of genes one can quantify simultaneously. To increase the throughput of this informative technique, a fluorescent bar-code system for the unique labeling of dozens of genes and an automated image analysis algorithm that enabled their simultaneous hybridization for the quantification of gene copy numbers were devised. We demonstrate the reliability of this multiplex approach on normal human lymphocytes, metaphase spreads of transformed cell lines, and cultured circulating tumor cells. It also opens the door to the development of gene panels for more comprehensive analysis of copy number changes in tissue, including the study of heterogeneity and of high-throughput clinical assays that could provide rapid quantification of gene copy numbers in samples with limited cellularity, such as circulating tumor cells.</P>
Dennis D. Keiser,JAN-FONG JUE,BRANDON D. MILLER,JIAN GAN,ADAM B. ROBINSON,PAVEL MEDVEDEV,JAMES MADDEN,DAN WACHS,MITCH MEYER 한국원자력학회 2014 Nuclear Engineering and Technology Vol.46 No.2
In order to investigate how the microstructure of fuel/matrix-interaction (FMI) layers change during irradiation, differentU–7Mo dispersion fuel plates have been irradiated to high fission density and then characterized using scanning electronmicroscopy (SEM). Specifially, samples from irradiated U–7Mo dispersion fuel elements with pure Al, Al–2Si and AA4043(~4.5 wt.%Si) matrices were SEM characterized using polished samples and samples that were prepared with a focused ionbeam (FIB). Features not observable for the polished samples could be captured in SEM images taken of the FIB samples. Forthe Al matrix sample, a relatively large FMI layer develops, with enrichment of Xe at the FMI layer/Al matrix interface andevidence of debonding. Overall, a significant penetration of Si from the FMI layer into the U–7Mo fuel was observed forsamples with Si in the Al matrix, which resulted in a change of the size (larger) and shape (round) of the fission gas bubbles. Additionally, solid fission product phases were observed to nucleate and grow within these bubbles. These changes in thelocalized regions of the microstructure of the U–7Mo may contribute to changes observed in the macroscopic swelling of fuelplates with Al–Si matrices.
Keiser, Dennis D. Jr.,Jue, Jan-Fong,Miller, Brandon D.,Gan, Jian,Robinson, Adam B.,Medvedev, Pavel,Madden, James,Wachs, Dan,Meyer, Mitch Korean Nuclear Society 2014 Nuclear Engineering and Technology Vol.46 No.2
In order to investigate how the microstructure of fuel/matrix-interaction (FMI) layers change during irradiation, different U-7Mo dispersion fuel plates have been irradiated to high fission density and then characterized using scanning electron microscopy (SEM). Specifially, samples from irradiated U-7Mo dispersion fuel elements with pure Al, Al-2Si and AA4043 (~4.5 wt.%Si) matrices were SEM characterized using polished samples and samples that were prepared with a focused ion beam (FIB). Features not observable for the polished samples could be captured in SEM images taken of the FIB samples. For the Al matrix sample, a relatively large FMI layer develops, with enrichment of Xe at the FMI layer/Al matrix interface and evidence of debonding. Overall, a significant penetration of Si from the FMI layer into the U-7Mo fuel was observed for samples with Si in the Al matrix, which resulted in a change of the size (larger) and shape (round) of the fission gas bubbles. Additionally, solid fission product phases were observed to nucleate and grow within these bubbles. These changes in the localized regions of the microstructure of the U-7Mo may contribute to changes observed in the macroscopic swelling of fuel plates with Al-Si matrices.