Combined Fluorescent-Chromogenic In Situ Hybridization for Identification and Laser Microdissection of Interphase Chromosomes

Chromosome territories constitute the most conspicuous feature of nuclear architecture, and they exhibit non-random distribution patterns in the interphase nucleus. We observed that in cell nuclei from humans with Down Syndrome two chromosomes 21 frequently localize proximal to one another and distant from the third chromosome. To systematically investigate whether the proximally positioned chromosomes were always the same in all cells, we developed an approach consisting of sequential FISH and CISH combined with laser-microdissection of chromosomes from the interphase nucleus and followed by subsequent chromosome identification by microsatellite allele genotyping. This approach identified proximally positioned chromosomes from cultured cells, and the analysis showed that the identity of the chromosomes proximally positioned varies. However, the data suggest that there may be a tendency of the same chromosomes to be positioned close to each other in the interphase nucleus of trisomic cells. The protocol described here represents a powerful new method for genome analysis

Comments on hydrographic and topographic LIDAR acquisition and merging with multibeam sounding data acquired in the Olympic Coast National Marine Sanctuary

In April 2005, a SHOALS 1000T LIDAR system was used as an efficient alternative for safely acquiring data to describe the existing conditions of nearshore bathymetry and the intertidal zone over an approximately 40.7 km2 (11.8 nm2) portion of hazardous coastline within the Olympic Coast National Marine Sanctuary (OCNMS). Data were logged from 1,593 km (860 nm) of track lines in just over 21 hours of flight time. Several islands and offshore rocks were also surveyed, and over 24,000 geo-referenced digital still photos were captured to assist with data cleaning and QA/QC. The 1 kHz bathymetry laser obtained a maximum water depth of 22.2 meters. Floating kelp beds, breaking surf lines and turbid water were all challenges to the survey. Although sea state was favorable for this time of the year, recent heavy rainfall and a persistent low-lying layer of fog reduced acquisition productivity. The existence of a completed VDatum model covering this same geographic region permitted the LIDAR data to be vertically transformed and merged with existing shallow water multibeam data and referenced to the mean lower low water (MLLW) tidal datum. Analysis of a multibeam bathymetry-LIDAR difference surface containing over 44,000 samples indicated surface deviations from –24.3 to 8.48 meters, with a mean difference of –0.967 meters, and standard deviation of 1.762 meters. Errors in data cleaning and false detections due to interference from surf, kelp, and turbidity likely account for the larger surface separations, while the remaining general surface difference trend could partially be attributed to a more dense data set, and shoal-biased cleaning, binning and gridding associated with the multibeam data for maintaining conservative least depths important for charting dangers to navigation. (PDF contains 27 pages.)