The Atmospheric Monitoring Strategy for the Cherenkov Telescope Array

The Imaging Atmospheric Cherenkov Technique (IACT) is unusual in astronomy as the atmosphere actually forms an intrinsic part of the detector system, with telescopes indirectly detecting very high energy particles by the generation and transport of Cherenkov photons deep within the atmosphere. This means that accurate measurement, characterisation and monitoring of the atmosphere is at the very heart of successfully operating an IACT system. The Cherenkov Telescope Array (CTA) will be the next generation IACT observatory with an ambitious aim to improve the sensitivity of an order of magnitude over current facilities, along with corresponding improvements in angular and energy resolution and extended energy coverage, through an array of Large (23m), Medium (12m) and Small (4m) sized telescopes spread over an area of order ~km$^2$. Whole sky coverage will be achieved by operating at two sites: one in the northern hemisphere and one in the southern hemisphere. This proceedings will cover the characterisation of the candidate sites and the atmospheric calibration strategy. CTA will utilise a suite of instrumentation and analysis techniques for atmospheric modelling and monitoring regarding pointing forecasts, intelligent pointing selection for the observatory operations and for offline data correction.Comment: 6 pages. To appear in the proceedings of the Adapting to the Atmosphere conference 201

Three-dimensional organization of chromosome territories in the human interphase nucleus

The synthesis of proteins, maintenance of structure and duplication of the eukaryotic cell itself are all fine-tuned biochemical processes that depend on the precise structural arrangement of the cellular components. The regulation of genes – their transcription and replication - has been shown to be connected closely to the three- dimensional organization of the genome in the cell nucleus. Despite the successful linear sequencing of the human genome its three-dimensional structure is widely unknown. The nucleus of the cell has for a long time been viewed as a 'spaghetti soup' of DNA bound to various proteins without much internal structure, except during cell division when chromosomes are condensed into separate entities. Only recently has it become apparent that chromosomes occupy distinct 'territories' also in the interphase, i.e. between cell divisions. In an analogy of the Bauhaus principle that "form follows function" we believe that analyzing in which form DNA is organized in these territories will help us to understand genomic function. We use computer models - Monte Carlo and Brownian dynamics simulations - to develop plausible proposals for the structure of the interphase genome and compare them to experimental data. In the work presented here, we simulate interphase chromosomes for different folding morphologies of the chromatin fiber which is organized into loops of 100kbp to 3 Mbp that can be interconnected in various ways. The backbone of the fiber is described by a wormlike-chain polymer whose diameter and stiffness can be estimated from independent measurements. The implementation describes this polymer as a segmented chain with 3000 to 20000 segments for chromosome 15 depending on the phase of the simulation. The modeling is performed on a parallel computer (IBM SP2 with 80 nodes). We also determine genomic marker distributions within the Prader- Willi-Region on chromosome 15q11.2-13.3. For these measurements we use a fluorescence in situ hybridisation method (in collaboration with I. Solovai, J. Craig and T. Cremer, Munich, FRG) conserving the structure of the nucleus. As probes we use 10 kbp long lambda clones (Prof. B. Horsthemke, Essen, FRG) covering genomic marker distances between 8 kbp and 250 kbp. The markers are detected with confocal and standing wavefield light microscopes (in collaboration with J.Rauch, J. Bradl, C. Cremer and E.Stelzer, both Heidelberg, FRG) and using special image reconstruction methods developed solely for this purpose (developed by R. Eils. and W. Jaeger, Heidelberg, FRG). Best agreement between simulations and experiments is reached for a Multi-Loop-Subcompartment model with a loop size of 126 kbp which are forming rosetts and are linked by a chromatin linker of 126 kbp. We also hypothesize a different folding structure for maternal versus paternal chromosome 15. In simulations of whole cell nuclei this modell also leads to distinct chromosome territories and subcompartments. A fractal analysis of the simulations leads to multifractal behavior in good agreement with predictions drawn from porous network research

Three-dimensional organization of chromosome territories in the human interphase cell nucleus.

The synthesis of proteins, maintenance of structure and duplication of the eukaryotic cell itself are all fine-tuned biochemical processes that depend on the precise structural arrangement of the cellular components. The regulation of genes – their transcription and replication - has been shown to be connected closely to the threedimensional organization of the genome in the cell nucleus. Despite the successful linear sequencing of the human genome its three-dimensional structure is widely unknown. The nucleus of the cell has for a long time been viewed as a 'spaghetti soup' of DNA bound to various proteins without much internal structure, except during cell division when chromosomes are condensed into separate entities. Only recently has it become apparent that chromosomes occupy distinct 'territories' also in the interphase, i.e. between cell divisions. In an analogy of the Bauhaus principle that "form follows function" we believe that analyzing in which form DNA is organized in these territories will help us to understand genomic function. We use computer models - Monte Carlo and Brownian dynamics simulations - to develop plausible proposals for the structure of the interphase genome and compare them to experimental data. In the work presented here, we simulate interphase chromosomes for different folding morphologies of the chromatin fiber which is organized into loops of 100kbp to 3 Mbp that can be interconnected in various ways. The backbone of the fiber is described by a wormlike-chain polymer whose diameter and stiffness can be estimated from independent measurements. The implementation describes this polymer as a segmented chain with 3000 to 20000 segments for chromosome 15 depending on the phase of the simulation. The modeling is performed on a parallel computer (IBM SP2 with 80 nodes). We also determine genomic marker distributions within the Prader- Willi-Region on chromosome 15q11.2-13.3. For these measurements we use a fluorescence in situ hybridisation method (in collaboration with I. Solovai, J. Craig and T. Cremer, Munich, FRG) conserving the structure of the nucleus. As probes we use 10 kbp long lambda clones (Prof. B. Horsthemke, Essen, FRG) covering genomic marker distances between 8 kbp and 250 kbp. The markers are detected with confocal and standing wavefield light microscopes (in collaboration with J.Rauch, J. Bradl, C. Cremer and E.Stelzer, both Heidelberg, FRG) and using special image reconstruction methods developed solely for this purpose (developed by R. Eils. and W. Jaeger, Heidelberg, FRG). Best agreement between simulations and experiments is reached for a Multi-Loop-Subcompartment model with a loop size of 126 kbp which are forming rosetts and are linked by a chromatin linker of 126 kbp. We also hypothesize a different folding structure for maternal versus paternal chromosome 15. In simulations of whole cell nuclei this modell also leads to distinct chromosome territories and subcompartments. A fractal analysis of the simulations leads to multifractal behavior in good agreement with predictions drawn from porous network research

Diffusion and transport in the human interphase cell nucleus - FCS experiments compared to simulations.

Despite the succesful linear sequencing of the human genome the three-dimensional arrangement of chromatin, functional, and structural components is still largely unknown. Molecular transport and diffusion are important for processes like gene regulation, replication, or repair and are vitally influenced by the structure. With a comparison between fluorescence correlation spectroscopy (FCS) experiments and simulations we show here an interdisciplinary approach for the understanding of transport and diffusion properties in the human interphase cell nucleus. For a long time the interphase nucleus has been viewed as a 'spaghetti soup' of DNA without much internal structure, except during cell division. Only recently has it become apparent that chromosomes occupy distinct 'territories' also in interphase. Two models for the detailed folding of the 30 nm chromatin fibre within these territories are under debate: In the Random-Walk/Giant-Loop-model big loops of 3 to 5 Mbp are attached to a non-DNA backbone. In the Multi-Loop-Subcompartment (MLS) model loops of around 120 kbp are forming rosettes which are also interconnected by the chromatin fibre. Here we show with a comparison between simulations and experiments an interdisciplinary approach leading to a determination of the three-dimensional organization of the human genome: For the predictions of experiments various models of human interphase chromosomes and the whole cell nucleus were simulated with Monte Carlo and Brownian Dynamics methods. Only the MLS-model leads to the formation of non-overlapping chromosome territories and distinct functional and dynamic subcompartments in agreement with experiments. Fluorescence in situ hybridization is used for the specific marking of chromosome arms and pairs of small chromosomal DNA regions. The labelling is visualized with confocal laser scanning microscopy followed by image reconstruction procedures. Chromosome arms show only small overlap and globular substructures as predicted by the MLS-model. The spatial distances between pairs of genomic markers as function of their genomic separation result in a MLS-model with loop and linker sizes around 126 kbp. With the development of GFP-fusion-proteins it is possible to study the chromatin distribution and dynamics resulting from cell cycle, treatment by chemicals or radiation in vivo. The chromatin distributions are similar to those found in the simulation of whole cell nuclei of the MLS-model. Fractal analysis is especially suited to quantify the unordered and non-euclidean chromatin distribution of the nucleus. The dynamic behaviour of the chromatin structure and the diffusion of particles in the nucleus are also closely connected to the fractal dimension. Fractal analysis of the simulations reveal the multi-fractality of chromosomes. First fractal analysis of chromatin distributions in vivo result in significant differences for different morphologies and might favour a MLS-model-like chromatin distribution. Simulations of fragment distributions based on double strand breakage after carbon-ion irradiation differ in different models. Here again a comparison with experiments favours a MLS-model. FCS in combination with a scanning device is a suitable tool to study the diffusion characteristics of fluorescent proteins in living cell nuclei with high spatial resolution. Computer simulations of the three-dimensional organization of the human interphase nucleus allows a detailed test of theoretical models in comparison to experiments. Diffusion and transport in the nucleus are most appropriately described with the concept of obstructed diffusion. A large volume fraction of the nucleus seems to contain a cytosol-like liquid with an apparent viscosity 5 times higher than in water. The geometry of particles and structure as well as their interactions influence the mobilities in terms of speed and spatial coverage. A considerable amount of genomic sites is accessible for not too large particles. FCS experiments and simulations based on the polymer model are in a good agreement. Using recently developed in vivo chromatin markers, a detailed study of mobility vs. structure is subject of current work