FocAn: automated 3D analysis of DNA repair foci in image stacks acquired by confocal fluorescence microscopy.

Abstract:

BACKGROUND:Phosphorylated histone H2AX, also known as γH2AX, forms μm-sized nuclear foci at the sites of DNA double-strand breaks (DSBs) induced by ionizing radiation and other agents. Due to their specificity and sensitivity, γH2AX immunoassays have become the gold standard for studying DSB induction and repair. One of these assays relies on the immunofluorescent staining of γH2AX followed by microscopic imaging and foci counting. During the last years, semi- and fully automated image analysis, capable of fast detection and quantification of γH2AX foci in large datasets of fluorescence images, are gradually replacing the traditional method of manual foci counting. A major drawback of the non-commercial software for foci counting (available so far) is that they are restricted to 2D-image data. In practice, these algorithms are useful for counting the foci located close to the midsection plane of the nucleus, while the out-of-plane foci are neglected. RESULTS:To overcome the limitations of 2D foci counting, we present a freely available ImageJ-based plugin (FocAn) for automated 3D analysis of γH2AX foci in z-image stacks acquired by confocal fluorescence microscopy. The image-stack processing algorithm implemented in FocAn is capable of automatic 3D recognition of individual cell nuclei and γH2AX foci, as well as evaluation of the total foci number per cell nucleus. The FocAn algorithm consists of two parts: nucleus identification and foci detection, each employing specific sequences of auto local thresholding in combination with watershed segmentation techniques. We validated the FocAn algorithm using fluorescence-labeled γH2AX in two glioblastoma cell lines, irradiated with 2 Gy and given up to 24 h post-irradiation for repair. We found that the data obtained with FocAn agreed well with those obtained with an already available software (FoCo) and manual counting. Moreover, FocAn was capable of identifying overlapping foci in 3D space, which ensured accurate foci counting even at high DSB density of up to ~ 200 DSB/nucleus. CONCLUSIONS:FocAn is freely available an open-source 3D foci analyzer. The user-friendly algorithm FocAn requires little supervision and can automatically count the amount of DNA-DSBs, i.e. fluorescence-labeled γH2AX foci, in 3D image stacks acquired by laser-scanning microscopes without additional nuclei staining.

journal_name

BMC Bioinformatics

journal_title

BMC bioinformatics

authors

Memmel S,Sisario D,Zimmermann H,Sauer M,Sukhorukov VL,Djuzenova CS,Flentje M

doi

10.1186/s12859-020-3370-8

subject

Has Abstract

pub_date

2020-01-28 00:00:00

pages

27

issue

1

issn

1471-2105

pii

10.1186/s12859-020-3370-8

journal_volume

21

pub_type

杂志文章
  • Effects of Mecp2 loss of function in embryonic cortical neurons: a bioinformatics strategy to sort out non-neuronal cells variability from transcriptome profiling.

    abstract:BACKGROUND:Mecp2 null mice model Rett syndrome (RTT) a human neurological disorder affecting females after apparent normal pre- and peri-natal developmental periods. Neuroanatomical studies in cerebral cortex of RTT mouse models revealed delayed maturation of neuronal morphology and autonomous as well as non-cell auton...

    journal_title:BMC bioinformatics

    pub_type: 杂志文章

    doi:10.1186/s12859-015-0859-7

    authors: Vacca M,Tripathi KP,Speranza L,Aiese Cigliano R,Scalabrì F,Marracino F,Madonna M,Sanseverino W,Perrone-Capano C,Guarracino MR,D'Esposito M

    更新日期:2016-01-20 00:00:00

  • Image-based classification of plant genus and family for trained and untrained plant species.

    abstract:BACKGROUND:Modern plant taxonomy reflects phylogenetic relationships among taxa based on proposed morphological and genetic similarities. However, taxonomical relation is not necessarily reflected by close overall resemblance, but rather by commonality of very specific morphological characters or similarity on the mole...

    journal_title:BMC bioinformatics

    pub_type: 杂志文章

    doi:10.1186/s12859-018-2474-x

    authors: Seeland M,Rzanny M,Boho D,Wäldchen J,Mäder P

    更新日期:2019-01-03 00:00:00

  • Colonyzer: automated quantification of micro-organism growth characteristics on solid agar.

    abstract:BACKGROUND:High-throughput screens comparing growth rates of arrays of distinct micro-organism cultures on solid agar are useful, rapid methods of quantifying genetic interactions. Growth rate is an informative phenotype which can be estimated by measuring cell densities at one or more times after inoculation. Precise ...

    journal_title:BMC bioinformatics

    pub_type: 杂志文章

    doi:10.1186/1471-2105-11-287

    authors: Lawless C,Wilkinson DJ,Young A,Addinall SG,Lydall DA

    更新日期:2010-05-28 00:00:00

  • Comparative evaluation of set-level techniques in predictive classification of gene expression samples.

    abstract:BACKGROUND:Analysis of gene expression data in terms of a priori-defined gene sets has recently received significant attention as this approach typically yields more compact and interpretable results than those produced by traditional methods that rely on individual genes. The set-level strategy can also be adopted wit...

    journal_title:BMC bioinformatics

    pub_type: 杂志文章

    doi:10.1186/1471-2105-13-S10-S15

    authors: Holec M,Kléma J,Zelezný F,Tolar J

    更新日期:2012-06-25 00:00:00

  • Cascaded classifiers for confidence-based chemical named entity recognition.

    abstract:BACKGROUND:Chemical named entities represent an important facet of biomedical text. RESULTS:We have developed a system to use character-based n-grams, Maximum Entropy Markov Models and rescoring to recognise chemical names and other such entities, and to make confidence estimates for the extracted entities. An adjusta...

    journal_title:BMC bioinformatics

    pub_type: 杂志文章

    doi:10.1186/1471-2105-9-S11-S4

    authors: Corbett P,Copestake A

    更新日期:2008-11-19 00:00:00

  • The exploration of disease-specific gene regulatory networks in esophageal carcinoma and stomach adenocarcinoma.

    abstract:BACKGROUND:Feed-forward loops (FFLs), consisting of miRNAs, transcription factors (TFs) and their common target genes, have been validated to be important for the initialization and development of complex diseases, including cancer. Esophageal Carcinoma (ESCA) and Stomach Adenocarcinoma (STAD) are two types of malignan...

    journal_title:BMC bioinformatics

    pub_type: 杂志文章

    doi:10.1186/s12859-019-3230-6

    authors: Qin G,Yang L,Ma Y,Liu J,Huo Q

    更新日期:2019-12-30 00:00:00

  • Compromise or optimize? The breakpoint anti-median.

    abstract:BACKGROUND:The median of k≥3 genomes was originally defined to find a compromise genome indicative of a common ancestor. However, in gene order comparisons, the usual definitions based on minimizing the sum of distances to the input genomes lead to degenerate medians reflecting only one of the input genomes. "Near-medi...

    journal_title:BMC bioinformatics

    pub_type: 杂志文章

    doi:10.1186/s12859-016-1340-y

    authors: Larlee CA,Brandts A,Sankoff D

    更新日期:2016-12-15 00:00:00

  • A Bayesian data fusion based approach for learning genome-wide transcriptional regulatory networks.

    abstract:BACKGROUND:Reverse engineering of transcriptional regulatory networks (TRN) from genomics data has always represented a computational challenge in System Biology. The major issue is modeling the complex crosstalk among transcription factors (TFs) and their target genes, with a method able to handle both the high number...

    journal_title:BMC bioinformatics

    pub_type: 杂志文章

    doi:10.1186/s12859-020-3510-1

    authors: Sauta E,Demartini A,Vitali F,Riva A,Bellazzi R

    更新日期:2020-05-29 00:00:00

  • DMDtoolkit: a tool for visualizing the mutated dystrophin protein and predicting the clinical severity in DMD.

    abstract:BACKGROUND:Dystrophinopathy is one of the most common human monogenic diseases which results in Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD). Mutations in the dystrophin gene are responsible for both DMD and BMD. However, the clinical phenotypes and treatments are quite different in these two m...

    journal_title:BMC bioinformatics

    pub_type: 杂志文章

    doi:10.1186/s12859-017-1504-4

    authors: Zhou J,Xin J,Niu Y,Wu S

    更新日期:2017-02-02 00:00:00

  • MACSIMS: multiple alignment of complete sequences information management system.

    abstract:BACKGROUND:In the post-genomic era, systems-level studies are being performed that seek to explain complex biological systems by integrating diverse resources from fields such as genomics, proteomics or transcriptomics. New information management systems are now needed for the collection, validation and analysis of the...

    journal_title:BMC bioinformatics

    pub_type: 杂志文章

    doi:10.1186/1471-2105-7-318

    authors: Thompson JD,Muller A,Waterhouse A,Procter J,Barton GJ,Plewniak F,Poch O

    更新日期:2006-06-23 00:00:00

  • Extended analysis of benchmark datasets for Agilent two-color microarrays.

    abstract:BACKGROUND:As part of its broad and ambitious mission, the MicroArray Quality Control (MAQC) project reported the results of experiments using External RNA Controls (ERCs) on five microarray platforms. For most platforms, several different methods of data processing were considered. However, there was no similar consid...

    journal_title:BMC bioinformatics

    pub_type: 杂志文章

    doi:10.1186/1471-2105-8-371

    authors: Kerr KF

    更新日期:2007-10-03 00:00:00

  • Hit integration for identifying optimal spaced seeds.

    abstract:BACKGROUND:Introduction of spaced speeds opened a way of sensitivity improvement in homology search without loss of search speed. Since then, the efforts of finding optimal seed which maximizes the sensitivity have been continued today. The sensitivity of a seed is generally computed by its hit probability. However, th...

    journal_title:BMC bioinformatics

    pub_type: 杂志文章

    doi:10.1186/1471-2105-11-S1-S37

    authors: Chung WH,Park SB

    更新日期:2010-01-18 00:00:00

  • Inferring functional modules of protein families with probabilistic topic models.

    abstract:BACKGROUND:Genome and metagenome studies have identified thousands of protein families whose functions are poorly understood and for which techniques for functional characterization provide only partial information. For such proteins, the genome context can give further information about their functional context. RESU...

    journal_title:BMC bioinformatics

    pub_type: 杂志文章

    doi:10.1186/1471-2105-12-141

    authors: Konietzny SG,Dietz L,McHardy AC

    更新日期:2011-05-09 00:00:00

  • iMEGES: integrated mental-disorder GEnome score by deep neural network for prioritizing the susceptibility genes for mental disorders in personal genomes.

    abstract:BACKGROUND:A range of rare and common genetic variants have been discovered to be potentially associated with mental diseases, but many more have not been uncovered. Powerful integrative methods are needed to systematically prioritize both variants and genes that confer susceptibility to mental diseases in personal gen...

    journal_title:BMC bioinformatics

    pub_type: 杂志文章

    doi:10.1186/s12859-018-2469-7

    authors: Khan A,Liu Q,Wang K

    更新日期:2018-12-28 00:00:00

  • A stepwise framework for the normalization of array CGH data.

    abstract:BACKGROUND:In two-channel competitive genomic hybridization microarray experiments, the ratio of the two fluorescent signal intensities at each spot on the microarray is commonly used to infer the relative amounts of the test and reference sample DNA levels. This ratio may be influenced by systematic measurement effect...

    journal_title:BMC bioinformatics

    pub_type: 杂志文章

    doi:10.1186/1471-2105-6-274

    authors: Khojasteh M,Lam WL,Ward RK,MacAulay C

    更新日期:2005-11-18 00:00:00

  • PDB-UF: database of predicted enzymatic functions for unannotated protein structures from structural genomics.

    abstract:BACKGROUND:The number of protein structures from structural genomics centers dramatically increases in the Protein Data Bank (PDB). Many of these structures are functionally unannotated because they have no sequence similarity to proteins of known function. However, it is possible to successfully infer function using o...

    journal_title:BMC bioinformatics

    pub_type: 杂志文章

    doi:10.1186/1471-2105-7-53

    authors: von Grotthuss M,Plewczynski D,Ginalski K,Rychlewski L,Shakhnovich EI

    更新日期:2006-02-06 00:00:00

  • Identification of germ cell-specific genes in mammalian meiotic prophase.

    abstract:BACKGROUND:Mammalian germ cells undergo meiosis to produce sperm or eggs, haploid cells that are primed to meet and propagate life. Meiosis is initiated by retinoic acid and meiotic prophase is the first and most complex stage of meiosis when homologous chromosomes pair to exchange genetic information. Errors in meiosi...

    journal_title:BMC bioinformatics

    pub_type: 杂志文章

    doi:10.1186/1471-2105-14-72

    authors: Li Y,Ray D,Ye P

    更新日期:2013-02-27 00:00:00

  • Fregene: simulation of realistic sequence-level data in populations and ascertained samples.

    abstract:BACKGROUND:FREGENE simulates sequence-level data over large genomic regions in large populations. Because, unlike coalescent simulators, it works forwards through time, it allows complex scenarios of selection, demography, and recombination to be modelled simultaneously. Detailed tracking of sites under selection is im...

    journal_title:BMC bioinformatics

    pub_type: 杂志文章

    doi:10.1186/1471-2105-9-364

    authors: Chadeau-Hyam M,Hoggart CJ,O'Reilly PF,Whittaker JC,De Iorio M,Balding DJ

    更新日期:2008-09-08 00:00:00

  • Multi-omic analysis of signalling factors in inflammatory comorbidities.

    abstract:BACKGROUND:Inflammation is a core element of many different, systemic and chronic diseases that usually involve an important autoimmune component. The clinical phase of inflammatory diseases is often the culmination of a long series of pathologic events that started years before. The systemic characteristics and relate...

    journal_title:BMC bioinformatics

    pub_type: 杂志文章

    doi:10.1186/s12859-018-2413-x

    authors: Xiao H,Bartoszek K,Lio' P

    更新日期:2018-11-30 00:00:00

  • Combining calls from multiple somatic mutation-callers.

    abstract:BACKGROUND:Accurate somatic mutation-calling is essential for insightful mutation analyses in cancer studies. Several mutation-callers are publicly available and more are likely to appear. Nonetheless, mutation-calling is still challenging and there is unlikely to be one established caller that systematically outperfor...

    journal_title:BMC bioinformatics

    pub_type: 杂志文章

    doi:10.1186/1471-2105-15-154

    authors: Kim SY,Jacob L,Speed TP

    更新日期:2014-05-21 00:00:00

  • πBUSS: a parallel BEAST/BEAGLE utility for sequence simulation under complex evolutionary scenarios.

    abstract:BACKGROUND:Simulated nucleotide or amino acid sequences are frequently used to assess the performance of phylogenetic reconstruction methods. BEAST, a Bayesian statistical framework that focuses on reconstructing time-calibrated molecular evolutionary processes, supports a wide array of evolutionary models, but lacked ...

    journal_title:BMC bioinformatics

    pub_type: 杂志文章

    doi:10.1186/1471-2105-15-133

    authors: Bielejec F,Lemey P,Carvalho LM,Baele G,Rambaut A,Suchard MA

    更新日期:2014-05-07 00:00:00

  • Ciruvis: a web-based tool for rule networks and interaction detection using rule-based classifiers.

    abstract:BACKGROUND:The use of classification algorithms is becoming increasingly important for the field of computational biology. However, not only the quality of the classification, but also its biological interpretation is important. This interpretation may be eased if interacting elements can be identified and visualized, ...

    journal_title:BMC bioinformatics

    pub_type: 杂志文章

    doi:10.1186/1471-2105-15-139

    authors: Bornelöv S,Marillet S,Komorowski J

    更新日期:2014-05-12 00:00:00

  • Ranking analysis of F-statistics for microarray data.

    abstract:BACKGROUND:Microarray technology provides an efficient means for globally exploring physiological processes governed by the coordinated expression of multiple genes. However, identification of genes differentially expressed in microarray experiments is challenging because of their potentially high type I error rate. Me...

    journal_title:BMC bioinformatics

    pub_type: 杂志文章

    doi:10.1186/1471-2105-9-142

    authors: Tan YD,Fornage M,Xu H

    更新日期:2008-03-06 00:00:00

  • Improving ontologies by automatic reasoning and evaluation of logical definitions.

    abstract:BACKGROUND:Ontologies are widely used to represent knowledge in biomedicine. Systematic approaches for detecting errors and disagreements are needed for large ontologies with hundreds or thousands of terms and semantic relationships. A recent approach of defining terms using logical definitions is now increasingly bein...

    journal_title:BMC bioinformatics

    pub_type: 杂志文章

    doi:10.1186/1471-2105-12-418

    authors: Köhler S,Bauer S,Mungall CJ,Carletti G,Smith CL,Schofield P,Gkoutos GV,Robinson PN

    更新日期:2011-10-27 00:00:00

  • Integrating gene expression and GO classification for PCA by preclustering.

    abstract:BACKGROUND:Gene expression data can be analyzed by summarizing groups of individual gene expression profiles based on GO annotation information. The mean expression profile per group can then be used to identify interesting GO categories in relation to the experimental settings. However, the expression profiles present...

    journal_title:BMC bioinformatics

    pub_type: 杂志文章

    doi:10.1186/1471-2105-11-158

    authors: De Haan JR,Piek E,van Schaik RC,de Vlieg J,Bauerschmidt S,Buydens LM,Wehrens R

    更新日期:2010-03-26 00:00:00

  • Systematic integration of experimental data and models in systems biology.

    abstract:BACKGROUND:The behaviour of biological systems can be deduced from their mathematical models. However, multiple sources of data in diverse forms are required in the construction of a model in order to define its components and their biochemical reactions, and corresponding parameters. Automating the assembly and use of...

    journal_title:BMC bioinformatics

    pub_type: 杂志文章

    doi:10.1186/1471-2105-11-582

    authors: Li P,Dada JO,Jameson D,Spasic I,Swainston N,Carroll K,Dunn W,Khan F,Malys N,Messiha HL,Simeonidis E,Weichart D,Winder C,Wishart J,Broomhead DS,Goble CA,Gaskell SJ,Kell DB,Westerhoff HV,Mendes P,Paton NW

    更新日期:2010-11-29 00:00:00

  • A multiple-alignment based primer design algorithm for genetically highly variable DNA targets.

    abstract:BACKGROUND:Primer design for highly variable DNA sequences is difficult, and experimental success requires attention to many interacting constraints. The advent of next-generation sequencing methods allows the investigation of rare variants otherwise hidden deep in large populations, but requires attention to populatio...

    journal_title:BMC bioinformatics

    pub_type: 杂志文章

    doi:10.1186/1471-2105-14-255

    authors: Brodin J,Krishnamoorthy M,Athreya G,Fischer W,Hraber P,Gleasner C,Green L,Korber B,Leitner T

    更新日期:2013-08-21 00:00:00

  • LAVA: an open-source approach to designing LAMP (loop-mediated isothermal amplification) DNA signatures.

    abstract:BACKGROUND:We developed an extendable open-source Loop-mediated isothermal AMPlification (LAMP) signature design program called LAVA (LAMP Assay Versatile Analysis). LAVA was created in response to limitations of existing LAMP signature programs. RESULTS:LAVA identifies combinations of six primer regions for basic LAM...

    journal_title:BMC bioinformatics

    pub_type: 杂志文章

    doi:10.1186/1471-2105-12-240

    authors: Torres C,Vitalis EA,Baker BR,Gardner SN,Torres MW,Dzenitis JM

    更新日期:2011-06-16 00:00:00

  • Efficient reconstruction of biological networks via transitive reduction on general purpose graphics processors.

    abstract:BACKGROUND:Techniques for reconstruction of biological networks which are based on perturbation experiments often predict direct interactions between nodes that do not exist. Transitive reduction removes such relations if they can be explained by an indirect path of influences. The existing algorithms for transitive re...

    journal_title:BMC bioinformatics

    pub_type: 杂志文章

    doi:10.1186/1471-2105-13-281

    authors: Bošnački D,Odenbrett MR,Wijs A,Ligtenberg W,Hilbers P

    更新日期:2012-10-30 00:00:00

  • An extensible six-step methodology to automatically generate fuzzy DSSs for diagnostic applications.

    abstract:BACKGROUND:The diagnosis of many diseases can be often formulated as a decision problem; uncertainty affects these problems so that many computerized Diagnostic Decision Support Systems (in the following, DDSSs) have been developed to aid the physician in interpreting clinical data and thus to improve the quality of th...

    journal_title:BMC bioinformatics

    pub_type: 杂志文章

    doi:10.1186/1471-2105-14-S1-S4

    authors: d'Acierno A,Esposito M,De Pietro G

    更新日期:2013-01-01 00:00:00