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Search Results: 1 - 10 of 3527 matches for " Stephan Preibisch "
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"Hook"-calibration of GeneChip-microarrays: Theory and algorithm
Hans Binder, Stephan Preibisch
Algorithms for Molecular Biology , 2008, DOI: 10.1186/1748-7188-3-12
Abstract: We present the so-called hook-calibration method which co-processes the log-difference (delta) and -sum (sigma) of the perfect match (PM) and mismatch (MM) probe-intensities. The MM probes are utilized as an internal reference which is subjected to the same hybridization law as the PM, however with modified characteristics. After sequence-specific affinity correction the method fits the Langmuir-adsorption model to the smoothed delta-versus-sigma plot. The geometrical dimensions of this so-called hook-curve characterize the particular hybridization in terms of simple geometric parameters which provide information about the mean non-specific background intensity, the saturation value, the mean PM/MM-sensitivity gain and the fraction of absent probes. This graphical summary spans a metrics system for expression estimates in natural units such as the mean binding constants and the occupancy of the probe spots. The method is single-chip based, i.e. it separately uses the intensities for each selected chip.The hook-method corrects the raw intensities for the non-specific background hybridization in a sequence-specific manner, for the potential saturation of the probe-spots with bound transcripts and for the sequence-specific binding of specific transcripts. The obtained chip characteristics in combination with the sensitivity corrected probe-intensity values provide expression estimates scaled in natural units which are given by the binding constants of the particular hybridization.The basic mechanism underlying the functioning of DNA microarrays is that of hybridization. Hybridization is defined as the binding between complementary single-stranded nucleic acids. In the case of microarrays one strand is anchored at the surface and the second one is dissolved in solution, referred to as probe and target, respectively. The experimental technique of detecting hybridized probes relies on the fluorescence intensity measurement to infer the transcript abundance specific for a
Specific and non specific hybridization of oligonucleotide probes on microarrays
Hans Binder,Stephan Preibisch
Quantitative Biology , 2004, DOI: 10.1529/biophysj.104.055343
Abstract: Gene expression analysis by means of microarrays is based on the sequence specific binding of mRNA to DNA oligonucleotide probes and its measurement using fluorescent labels. The binding of RNA fragments involving other sequences than the intended target is problematic because it adds a "chemical background" to the signal, which is not related to the expression degree of the target gene. The paper presents a molecular signature of specific and non specific hybridization with potential consequences for gene expression analysis. We analyzed the signal intensities of perfect match (PM) and mismatch (MM) probes of GeneChip microarrays to specify the effect of specific and non specific hybridization. We found that these events give rise to different relations between the PM and MM intensities as function of the middle base of the PMs, namely a triplet- (C>G=T>A>0) and a duplet-like (C=T>0>G=A) pattern of the PM-MM log-intensity difference upon binding of specific and non specific RNA fragments, respectively. The systematic behaviour of the intensity difference can be rationalized on the level of base pairings of DNA/RNA oligonucleotide duplexes in the middle of the probe sequence. Non-specific binding is characterized by the reversal of the central Watson Crick (WC) pairing for each PM/MM probe pair, whereas specific binding refers to the combination of a WC and a self complementary (SC) pairing in PM and MM probes, respectively. The intensity of complementary MM introduces a systematic source of variation which decreases the precision of expression measures based on the MM intensities.
"Hook"-calibration of GeneChip-microarrays: Chip characteristics and expression measures
Hans Binder, Knut Krohn, Stephan Preibisch
Algorithms for Molecular Biology , 2008, DOI: 10.1186/1748-7188-3-11
Abstract: In this publication we summarize the essential chip characteristics provided by this method, analyze special benchmark experiments to estimate transcript related expression measures and illustrate the potency of the method to detect and to quantify the quality of a particular hybridization. It is shown that our single-chip approach provides expression measures responding linearly on changes of the transcript concentration over three orders of magnitude. In addition, the method calculates a detection call judging the relation between the signal and the detection limit of the particular measurement. The performance of the method in the context of different chip generations and probe set assignments is illustrated. The hook method characterizes the RNA-quality in terms of the 3'/5'-amplification bias and the sample-specific calling rate. We show that the proper judgement of these effects requires the disentanglement of non-specific and specific hybridization which, otherwise, can lead to misinterpretations of expression changes. The consequences of modifying probe/target interactions by either changing the labelling protocol or by substituting RNA by DNA targets are demonstrated.The single-chip based hook-method provides accurate expression estimates and chip-summary characteristics using the natural metrics given by the hybridization reaction with the potency to develop new standards for microarray quality control and calibration.DNA microarray technology enables conducting experiments that measure RNA-transcript abundance (so called gene expression or expression degree) on a large scale of genomic sequences. The quality of the measurement systematically depends on experimental factors such as the performance of the measuring "device", e.g., on the chosen array-type, the design of the chip-platform and -generation and on the particular probe design, on one hand; and also on the quality of the sample, e.g. on the source of RNA and the used hybridization-pipeline includ
Base pair interactions and hybridization isotherms of matched and mismatched oligonucleotide probes on microarrays
Hans Binder,Stephan Preibisch,Toralf Kirsten
Quantitative Biology , 2005,
Abstract: The lack of specificity in microarray experiments due to non-specific hybridization raises a serious problem for the analysis of microarray data because the residual chemical background intensity is not related to the expression degree of the gene of interest. We analyzed the concentration dependence of the signal intensity of perfect match (PM) and mismatch (MM) probes in terms using a microscopic binding model using a combination of mean hybridization isotherms and single base related affinity terms. The signal intensities of the PM and MM probes and their difference are assessed with regard to their sensitivity, specificity and resolution for gene expression measures. The presented theory implies the refinement of existing algorithms of probe level analysis to correct microarray data for non-specific background intensities.
BigDataViewer: Interactive Visualization and Image Processing for Terabyte Data Sets
Tobias Pietzsch,Stephan Saalfeld,Stephan Preibisch,Pavel Tomancak
Computer Science , 2014,
Abstract: The increasingly popular light sheet microscopy techniques generate very large 3D time-lapse recordings of living biological specimen. The necessity to make large volumetric datasets available for interactive visualization and analysis has been widely recognized. However, existing solutions build on dedicated servers to generate virtual slices that are transferred to the client applications, practically leading to insufficient frame rates (less than 10 frames per second) for truly interactive experience. An easily accessible open source solution for interactive arbitrary virtual re-slicing of very large volumes and time series of volumes has yet been missing. We fill this gap with BigDataViewer, a Fiji plugin to interactively navigate and visualize large image sequences from both local and remote data sources.
An automated workflow for parallel processing of large multiview SPIM recordings
Christopher Schmied,Peter Steinbach,Tobias Pietzsch,Stephan Preibisch,Pavel Tomancak
Quantitative Biology , 2015,
Abstract: Multiview light sheet fluorescence microscopy (LSFM) allows to image developing organisms in 3D at unprecedented temporal resolution over long periods of time. The resulting massive amounts of raw image data requires extensive processing interactively via dedicated graphical user interface (GUI) applications. The consecutive processing steps can be easily automated and the individual time points can be processed independently, which lends itself to trivial parallelization on a high performance cluster (HPC). Here we introduce an automated workflow for processing large multiview, multi-channel, multi-illumination time-lapse LSFM data on a single workstation or in parallel on a HPC. The pipeline relies on snakemake to resolve dependencies among consecutive processing steps and can be easily adapted to any cluster environment for processing LSFM data in a fraction of the time required to collect it.
An Integrated Micro- and Macroarchitectural Analysis of the Drosophila Brain by Computer-Assisted Serial Section Electron Microscopy
Albert Cardona,Stephan Saalfeld,Stephan Preibisch,Benjamin Schmid,Anchi Cheng,Jim Pulokas,Pavel Tomancak,Volker Hartenstein
PLOS Biology , 2012, DOI: 10.1371/journal.pbio.1000502
Abstract: The analysis of microcircuitry (the connectivity at the level of individual neuronal processes and synapses), which is indispensable for our understanding of brain function, is based on serial transmission electron microscopy (TEM) or one of its modern variants. Due to technical limitations, most previous studies that used serial TEM recorded relatively small stacks of individual neurons. As a result, our knowledge of microcircuitry in any nervous system is very limited. We applied the software package TrakEM2 to reconstruct neuronal microcircuitry from TEM sections of a small brain, the early larval brain of Drosophila melanogaster. TrakEM2 enables us to embed the analysis of the TEM image volumes at the microcircuit level into a light microscopically derived neuro-anatomical framework, by registering confocal stacks containing sparsely labeled neural structures with the TEM image volume. We imaged two sets of serial TEM sections of the Drosophila first instar larval brain neuropile and one ventral nerve cord segment, and here report our first results pertaining to Drosophila brain microcircuitry. Terminal neurites fall into a small number of generic classes termed globular, varicose, axiform, and dendritiform. Globular and varicose neurites have large diameter segments that carry almost exclusively presynaptic sites. Dendritiform neurites are thin, highly branched processes that are almost exclusively postsynaptic. Due to the high branching density of dendritiform fibers and the fact that synapses are polyadic, neurites are highly interconnected even within small neuropile volumes. We describe the network motifs most frequently encountered in the Drosophila neuropile. Our study introduces an approach towards a comprehensive anatomical reconstruction of neuronal microcircuitry and delivers microcircuitry comparisons between vertebrate and insect neuropile.
An Integrated Micro- and Macroarchitectural Analysis of the Drosophila Brain by Computer-Assisted Serial Section Electron Microscopy
Albert Cardona,Stephan Saalfeld,Stephan Preibisch,Benjamin Schmid,Anchi Cheng,Jim Pulokas,Pavel Tomancak,Volker Hartenstein
PLOS Biology , 2010, DOI: 10.1371/journal.pbio.1000502
Abstract: The analysis of microcircuitry (the connectivity at the level of individual neuronal processes and synapses), which is indispensable for our understanding of brain function, is based on serial transmission electron microscopy (TEM) or one of its modern variants. Due to technical limitations, most previous studies that used serial TEM recorded relatively small stacks of individual neurons. As a result, our knowledge of microcircuitry in any nervous system is very limited. We applied the software package TrakEM2 to reconstruct neuronal microcircuitry from TEM sections of a small brain, the early larval brain of Drosophila melanogaster. TrakEM2 enables us to embed the analysis of the TEM image volumes at the microcircuit level into a light microscopically derived neuro-anatomical framework, by registering confocal stacks containing sparsely labeled neural structures with the TEM image volume. We imaged two sets of serial TEM sections of the Drosophila first instar larval brain neuropile and one ventral nerve cord segment, and here report our first results pertaining to Drosophila brain microcircuitry. Terminal neurites fall into a small number of generic classes termed globular, varicose, axiform, and dendritiform. Globular and varicose neurites have large diameter segments that carry almost exclusively presynaptic sites. Dendritiform neurites are thin, highly branched processes that are almost exclusively postsynaptic. Due to the high branching density of dendritiform fibers and the fact that synapses are polyadic, neurites are highly interconnected even within small neuropile volumes. We describe the network motifs most frequently encountered in the Drosophila neuropile. Our study introduces an approach towards a comprehensive anatomical reconstruction of neuronal microcircuitry and delivers microcircuitry comparisons between vertebrate and insect neuropile.
Efficient Bayesian-based Multi-View Deconvolution
Stephan Preibisch,Fernando Amat,Evangelia Stamataki,Mihail Sarov,Robert H. Singer,Eugene Myers,Pavel Tomancak
Quantitative Biology , 2013, DOI: 10.1038/nmeth.2929
Abstract: Light sheet fluorescence microscopy is able to image large specimen with high resolution by imaging the sam- ples from multiple angles. Multi-view deconvolution can significantly improve the resolution and contrast of the images, but its application has been limited due to the large size of the datasets. Here we present a Bayesian- based derivation of multi-view deconvolution that drastically improves the convergence time and provide a fast implementation utilizing graphics hardware.
OpenSPIM - an open access platform for light sheet microscopy
Peter Pitrone,Johannes Schindelin,Luke Stuyvenberg,Stephan Preibisch,Michael Weber,Kevin W. Eliceiri,Jan Huisken,Pavel Tomancak
Quantitative Biology , 2013, DOI: 10.1038/nmeth.2507
Abstract: Light sheet microscopy promises to revolutionize developmental biology by enabling live in toto imaging of entire embryos with minimal phototoxicity. We present detailed instructions for building a compact and customizable Selective Plane Illumination Microscopy (SPIM) system. The integrated OpenSPIM hardware and software platform is shared with the scientific community through a public website, thereby making light sheet microscopy accessible for widespread use and optimization to various applications.
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