Orientation-independent reaction activity monitoring with single particle and data analytics

2021 ◽  
Vol 590 ◽  
pp. 458-466
Author(s):  
Jun Zhou ◽  
Zi Yu Pan ◽  
Hui Liu ◽  
Peng Fei Gao ◽  
Cheng Zhi Huang
Author(s):  
J. Frank ◽  
P.-Y. Sizaret ◽  
A. Verschoor ◽  
J. Lamy

The accuracy with which the attachment site of immunolabels bound to macromolecules may be localized in electron microscopic images can be considerably improved by using single particle averaging. The example studied in this work showed that the accuracy may be better than the resolution limit imposed by negative staining (∽2nm).The structure used for this demonstration was a halfmolecule of Limulus polyphemus (LP) hemocyanin, consisting of 24 subunits grouped into four hexamers. The top view of this structure was previously studied by image averaging and correspondence analysis. It was found to vary according to the flip or flop position of the molecule, and to the stain imbalance between diagonally opposed hexamers (“rocking effect”). These findings have recently been incorporated into a model of the full 8 × 6 molecule.LP hemocyanin contains eight different polypeptides, and antibodies specific for one, LP II, were used. Uranyl acetate was used as stain. A total of 58 molecule images (29 unlabelled, 29 labelled with antl-LPII Fab) showing the top view were digitized in the microdensitometer with a sampling distance of 50μ corresponding to 6.25nm.


Author(s):  
Adriana Verschoor ◽  
Ronald Milligan ◽  
Suman Srivastava ◽  
Joachim Frank

We have studied the eukaryotic ribosome from two vertebrate species (rabbit reticulocyte and chick embryo ribosomes) in several different electron microscopic preparations (Fig. 1a-d), and we have applied image processing methods to two of the types of images. Reticulocyte ribosomes were examined in both negative stain (0.5% uranyl acetate, in a double-carbon preparation) and frozen hydrated preparation as single-particle specimens. In addition, chick embryo ribosomes in tetrameric and crystalline assemblies in frozen hydrated preparation have been examined. 2D averaging, multivariate statistical analysis, and classification methods have been applied to the negatively stained single-particle micrographs and the frozen hydrated tetramer micrographs to obtain statistically well defined projection images of the ribosome (Fig. 2a,c). 3D reconstruction methods, the random conical reconstruction scheme and weighted back projection, were applied to the negative-stain data, and several closely related reconstructions were obtained. The principal 3D reconstruction (Fig. 2b), which has a resolution of 3.7 nm according to the differential phase residual criterion, can be compared to the images of individual ribosomes in a 2D tetramer average (Fig. 2c) at a similar resolution, and a good agreement of the general morphology and of many of the characteristic features is seen.Both data sets show the ribosome in roughly the same ’view’ or orientation, with respect to the adsorptive surface in the electron microscopic preparation, as judged by the agreement in both the projected form and the distribution of characteristic density features. The negative-stain reconstruction reveals details of the ribosome morphology; the 2D frozen-hydrated average provides projection information on the native mass-density distribution within the structure. The 40S subunit appears to have an elongate core of higher density, while the 60S subunit shows a more complex pattern of dense features, comprising a rather globular core, locally extending close to the particle surface.


2009 ◽  
Vol 14 (2) ◽  
pp. 142-152 ◽  
Author(s):  
Johannes B.J. Bussmann ◽  
Ulrich W. Ebner-Priemer ◽  
Jochen Fahrenberg

Behavior is central to psychology in almost any definition. Although observable activity is a core aspect of behavior, assessment strategies have tended to focus on emotional, cognitive, or physiological responses. When physical activity is assessed, it is done so mostly with questionnaires. Converging evidence of only a moderate association between self-reports of physical activity and objectively measured physical activity does raise questions about the validity of these self-reports. Ambulatory activity monitoring, defined as the measurement strategy to assess physical activity, posture, and movement patterns continuously in everyday life, has made major advances over the last decade and has considerable potential for further application in the assessment of observable activity, a core aspect of behavior. With new piezoresistive sensors and advanced computer algorithms, the objective measurement of physical activity, posture, and movement is much more easily achieved and measurement precision has improved tremendously. With this overview, we introduce to the reader some recent developments in ambulatory activity monitoring. We will elucidate the discrepancies between objective and subjective reports of activity, outline recent methodological developments, and offer the reader a framework for developing insight into the state of the art in ambulatory activity-monitoring technology, discuss methodological aspects of time-based design and psychometric properties, and demonstrate recent applications. Although not yet main stream, ambulatory activity monitoring – especially in combination with the simultaneous assessment of emotions, mood, or physiological variables – provides a comprehensive methodology for psychology because of its suitability for explaining behavior in context.


1998 ◽  
Vol 08 (PR6) ◽  
pp. Pr6-109-Pr6-113
Author(s):  
P. Gallo ◽  
F. Sciortino ◽  
P. Tartaglia ◽  
S.-H. Chen

1987 ◽  
Vol 48 (C5) ◽  
pp. C5-581-C5-584
Author(s):  
G. FASOL ◽  
N. MESTRES ◽  
M. DOBERS ◽  
A. FISCHER ◽  
K. PLOOG

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