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Cryo-electron tomography provides a snapshot of the cellular proteome. With template matching, the spatial positions of various macromolecular complexes within their native cellular context can be detected. However, the growing awareness of the reference bias introduced by the cross-correlation based approaches, and more importantly the lack of a reliable confidence measurement in the selection of these macromolecular complexes, has restricted the use of these applications. Here we propose a heuristic, in which the reference bias is measured in real space in an analogous way to the R-free value in X-ray crystallography. We measure the reference bias within the mask used to outline the area of the template, and do not modify the template itself. The heuristic works by splitting the mask into a working and a testing area in a volume ratio of 9:1. While the working area is used during the calculation of the cross-correlation function, the information from both areas is explored to calculate the M-free score. We show using artificial data, that the M-free score gives a reliable measure for the reference bias. The heuristic can be applied in template matching and in sub-tomogram averaging. We further test the applicability of the heuristic in tomograms of purified macromolecules, and tomograms of whole Mycoplasma cells.
The genus Casmaria H. Adams & A. Adams, 1853 (family Cassidae) is widespread in the tropical Indo-Pacific and has been documented from some Atlantic localities as well. Two Casmaria species, C. erinaceus (Linnaeus, 1758) and C. ponderosa (Gmelin, 1791), are common in Indo-Pacific shallow-water sandy bottom communities and are characterized by high morphological variability; both species encompass multiple, often sympatric forms of uncertain status. In the present study we carry out a phylogenetic analysis of some Philippine Casmaria morphs and demonstrate that one of the distinctive morphs earlier assigned to Casmaria ponderosa is in fact a different species, which we describe as Casmaria boblehmani sp. nov. The smooth form of Casmaria ponderosa, C. ponderosa ponderosa, and the solid nodulose form, widely called “form nodulosa” despite being strikingly different in shell morphology, are shown to be conspecific. Studied specimens of these two morphs even from different localities share the same haplotype of the CO1 gene. In light of these new data on the morphological variability of Casmaria species, we discuss criteria of species delimitation in the genus Casmaria and possible affinities of Casmaria boblehmani sp. nov. within the genus.
Correlative microscopy incorporates the specificity of fluorescent protein labeling into high-resolution electron micrographs. Several approaches exist for correlative microscopy, most of which have used the green fluorescent protein (GFP) as the label for light microscopy. Here we use chemical tagging and synthetic fluorophores instead, in order to achieve protein-specific labeling, and to perform multicolor imaging. We show that synthetic fluorophores preserve their post-embedding fluorescence in the presence of uranyl acetate. Post-embedding fluorescence is of such quality that the specimen can be prepared with identical protocols for scanning electron microscopy (SEM) and transmission electron microscopy (TEM); this is particularly valuable when singular or otherwise difficult samples are examined. We show that synthetic fluorophores give bright, well-resolved signals in super-resolution light microscopy, enabling us to superimpose light microscopic images with a precision of up to 25 nm in the x-y plane on electron micrographs. To exemplify the preservation quality of our new method we visualize the molecular arrangement of cadherins in adherens junctions of mouse epithelial cells.
Physical Biology is a field of life sciences dealing with the extraction of quantitative data from biophysical or molecular biological experiments with different levels of complexity. Such data are further used as parameters for mathematical models of the biological system. These models allow to predict reactions on external stimuli by describing the relevant molecular interactions and are therefore used for example to generate a deeper comprehension of complex human diseases. An essential technique in biophysical research on human diseases is fluorescence microscopy. This is a constantly developed toolbox comprising a large number of specific labeling strategies, as well as a broad spectrum of fluorescent probes. It is further minimal invasive and therefore suitable for measurements in living cells or organisms. The sensitivity of modern photo-detectors even allows for the detection of a single fluorescent probe with an accuracy of approximately 10 nm.
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The model-prediction was further verified by two color SMLM experiments. In this work the development and application of imaging-systems are described which provide quantitative data with single-molecule resolution for systems biological model approaches with a low degree of abstractness. In the near future, the impact of mathematical models in the research field of complex human diseases will increase. The predictions of these models will be more exact, the more detailed and accurate the input parameters will become. This work gives an impression of how quantitative data obtained by SMLM may serve as input parameters for mathematical models at the single-cell level.
Aufgrund der §§ 20, 44 Abs. 1 Nr. 1 des Hessischen Hochschulgesetztes in der Fassung vom 14. Dezember 2009, zuletzt geändert durch Gesetz vom 27. Mai 2013, hat der Fachbereichsrat des Fachbereichs Wirtschaftswissenschaf-ten der Johann Wolfgang Goethe-Universität Frankfurt am Main am 16. Juli 2014 die folgende Ordnung für den Masterstudiengang Betriebswirtschaftslehre beschlossen. Diese Ordnung hat das Präsidium der Johann Wolfgang Goethe-Universität gemäß § 37 Abs. 5 Hessisches Hochschulgesetz am 5. August 2014 genehmigt. Sie wird hiermit bekannt gegeben.
Aufgrund des Beschlusses des Fachbereichsrats des Fachbereichs Evangelische Theologie (FB 6) vom 19. Juni 2013, des Fachbereichsrats des Fachbereichs Sprach- und Kulturwissenschaften (FB 9) vom 24. April 2013 und 22. Mai 2013, des Fachbereichsrats des Fachbereichs Neuere Philologien (FB 10) und eines Umlaufverfahrens der an dem Abschluss Doktor der Philosophie/Doktorin der Philosophie (Dr. phil.) beteiligten Fachbereiche 3 - 11 im August/ September/Oktober 2013 wird die Ordnung zur Erlangung des akademischen Grades eines Doktors der Philosophie (Dr. phil.) oder einer Doktorin der Philosophie (Dr. phil.) vom 26. Juni 2001 (Staatsanzeiger Nr. 46/2001, S. 4.026 ff.) in der Fassung vom 7. August 2013 wie folgt geändert: