scholarly journals Building Accurate Intracellular Polarity Maps through Multiparametric Microscopy

2020 ◽  
Vol 3 (4) ◽  
pp. 78
Author(s):  
M. Carmen Gonzalez-Garcia ◽  
Pilar Herrero-Foncubierta ◽  
Emilio Garcia-Fernandez ◽  
Angel Orte

The precise knowledge of intracellular polarity, a physiological parameter that involves complex and intertwined intracellular mechanisms, may be relevant in the study of important diseases like cancer or Alzheimer’s. In this technical note, we illustrate our recently developed, accurate method for obtaining intracellular polarity maps employing potent fluorescence microscopy techniques. Our method is based on the selection of appropriate luminescent probes, in which several emission properties vary with microenvironment polarity, specifically spectral shifts and luminescence lifetime. A multilinear calibration is performed, correlating polarity vs. spectral shift vs. luminescence lifetime, to generate a powerful and error-free 3D space for reliable interpolation of microscopy data. Multidimensional luminescence microscopy is then used to obtain simultaneously spectral shift and luminescence lifetime images, which are then interpolated in the 3D calibration space, resulting in accurate, quantitative polarity maps.

2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Orkun Furat ◽  
Lukas Petrich ◽  
Donal P. Finegan ◽  
David Diercks ◽  
Francois Usseglio-Viretta ◽  
...  

AbstractAccurately capturing the architecture of single lithium-ion electrode particles is necessary for understanding their performance limitations and degradation mechanisms through multi-physics modeling. Information is drawn from multimodal microscopy techniques to artificially generate LiNi0.5Mn0.3Co0.2O2 particles with full sub-particle grain detail. Statistical representations of particle architectures are derived from X-ray nano-computed tomography data supporting an ‘outer shell’ model, and sub-particle grain representations are derived from focused-ion beam electron backscatter diffraction data supporting a ‘grain’ model. A random field model used to characterize and generate the outer shells, and a random tessellation model used to characterize and generate grain architectures, are combined to form a multi-scale model for the generation of virtual electrode particles with full-grain detail. This work demonstrates the possibility of generating representative single electrode particle architectures for modeling and characterization that can guide synthesis approaches of particle architectures with enhanced performance.


2013 ◽  
Vol 13 (12) ◽  
pp. 31607-31634 ◽  
Author(s):  
M. Rex ◽  
S. Kremser ◽  
P. Huck ◽  
G. Bodeker ◽  
I. Wohltmann ◽  
...  

Abstract. An extremely fast model to estimate the degree of stratospheric ozone depletion during polar winters is described. It is based on a set of coupled differential equations that simulate the seasonal evolution of vortex-averaged hydrogen chloride (HCl), nitric acid (HNO3), chlorine nitrate (ClONO2), active forms of chlorine (ClOx = Cl + ClO + 2 ClOOCl) and ozone (O3) on isentropic levels within the polar vortices. Terms in these equations account for the chemical and physical processes driving the time rate of change of these species. Eight empirical fit coefficients associated with these terms are derived by iteratively fitting the equations to vortex-averaged satellite-based measurements of HCl, HNO3 and ClONO2 and observationally derived ozone loss rates. The system of differential equations is not stiff and can be solved with a time step of one day, allowing many years to be processed per second on a standard PC. The inputs required are the daily fractions of the vortex area covered by polar stratospheric clouds and the fractions of the vortex area exposed to sunlight. The resultant model, SWIFT (Semi-empirical Weighted Iterative Fit Technique), provides a fast yet accurate method to simulate ozone loss rates in polar regions. SWIFT's capabilities are demonstrated by comparing measured and modeled total ozone loss outside of the training period.


2010 ◽  
Vol 16 (1) ◽  
pp. 64-72 ◽  
Author(s):  
David Baddeley ◽  
Mark B. Cannell ◽  
Christian Soeller

AbstractLocalization microscopy techniques based on localizing single fluorophore molecules now routinely achieve accuracies better than 30 nm. Unlike conventional optical microscopies, localization microscopy experiments do not generate an image but a list of discrete coordinates of estimated fluorophore positions. Data display and analysis therefore generally require visualization methods that translate the position data into conventional images. Here we investigate the properties of several widely used visualization techniques and show that a commonly used algorithm based on rendering Gaussians may lead to a 1.44-fold loss of resolution. Existing methods typically do not explicitly take sampling considerations into account and thus may produce spurious structures. We present two additional visualization algorithms, an adaptive histogram method based on quad-trees and a Delaunay triangulation based visualization of point data that address some of these deficiencies. The new visualization methods are designed to suppress erroneous detail in poorly sampled image areas but avoid loss of resolution in well-sampled regions. A number of criteria for scoring visualization methods are developed as a guide for choosing among visualization methods and are used to qualitatively compare various algorithms.


2004 ◽  
Vol 820 ◽  
Author(s):  
Z. Huang ◽  
D.A. Dikin ◽  
W. Ding ◽  
Y. Qiao ◽  
Y. Fridman ◽  
...  

AbstractNanostructures, such as nanowires, nanotubes, and nanocoils, can be described in many cases as quasi one-dimensional (1D) curved objects projecting in three-dimensional (3D) space. A parallax method to reconstruct the correct three-dimensional geometry of such 1D nanostructures is presented. A series of images were acquired at different view angles, and from those image pairs, 3D representations were constructed using a MATLAB program. Error analysis as a function of view-angle between the two images is discussed. As an example application, we demonstrate the importance of knowing the true 3D shape of Boron nanowires. Without precise knowledge of the nanowire's dimensions, diameter and length, mechanical resonance data cannot be properly fit to obtain an accurate estimate of the Young's modulus.


2014 ◽  
Vol 14 (13) ◽  
pp. 6545-6555 ◽  
Author(s):  
M. Rex ◽  
S. Kremser ◽  
P. Huck ◽  
G. Bodeker ◽  
I. Wohltmann ◽  
...  

Abstract. An extremely fast model to estimate the degree of stratospheric ozone depletion during polar winters is described. It is based on a set of coupled differential equations that simulate the seasonal evolution of vortex-averaged hydrogen chloride (HCl), nitric acid (HNO3), chlorine nitrate (ClONO2), active forms of chlorine (ClOx = Cl + ClO + 2 ClOOCl) and ozone (O3) on isentropic levels within the polar vortices. Terms in these equations account for the chemical and physical processes driving the time rate of change of these species. Eight empirical fit coefficients associated with these terms are derived by iteratively fitting the equations to vortex-averaged satellite-based measurements of HCl, HNO3 and ClONO2 and observationally derived ozone loss rates. The system of differential equations is not stiff and can be solved with a time step of one day, allowing many years to be processed per second on a standard PC. The inputs required are the daily fractions of the vortex area covered by polar stratospheric clouds and the fractions of the vortex area exposed to sunlight. The resultant model, SWIFT (Semi-empirical Weighted Iterative Fit Technique), provides a fast yet accurate method to simulate ozone loss rates in polar regions. SWIFT's capabilities are demonstrated by comparing measured and modeled total ozone loss outside of the training period.


2015 ◽  
pp. S1-S10
Author(s):  
E. KOZIOLOVA ◽  
O. JANOUSKOVA ◽  
P. CHYTIL ◽  
M. STUDENOVSKY ◽  
L. KOSTKA ◽  
...  

Anthracyclines, e.g. doxorubicin, pirarubicin, are widely used as cytostatic agents in the polymer nanotherapeutics designed for the highly effective antitumor therapy with reduced side effects. However, their precise dosage scheme needs to be optimized, which requires an accurate method for their quantification on the cellular level in vitro during nanocarrier development and in body fluids and tissues during testing in vivo. Various methods detecting the anthracycline content in biological samples have already been designed. Most of them are highly demanding and they differ in exactness and reproducibility. The cellular uptake and localization is predominantly observed and determined by microscopy techniques, the anthracycline content is usually quantified by chromatographic analysis using fluorescence detection. We reviewed and compared published methods concerning the detection of anthracycline nanocarriers.


Author(s):  
Chester J. Calbick ◽  
Richard E. Hartman

Quantitative studies of the phenomenon associated with reactions induced by the electron beam between specimens and gases present in the electron microscope require precise knowledge and control of the local environment experienced by the portion of the specimen in the electron beam. Because of outgassing phenomena, the environment at the irradiated portion of the specimen is very different from that in any place where gas pressures and compositions can be measured. We have found that differential pumping of the specimen chamber by a 4" Orb-Ion pump, following roughing by a zeolite sorption pump, can produce a specimen-chamber pressure 100- to 1000-fold less than that in the region below the objective lens.


Author(s):  
S. R. Herd ◽  
P. Chaudhari

Electron diffraction and direct transmission have been used extensively to study the local atomic arrangement in amorphous solids and in particular Ge. Nearest neighbor distances had been calculated from E.D. profiles and the results have been interpreted in terms of the microcrystalline or the random network models. Direct transmission electron microscopy appears the most direct and accurate method to resolve this issue since the spacial resolution of the better instruments are of the order of 3Å. In particular the tilted beam interference method is used regularly to show fringes corresponding to 1.5 to 3Å lattice planes in crystals as resolution tests.


Author(s):  
C.E. Voegele-Kliewer ◽  
A.D. McMaster ◽  
G.W. Dirks

Materials other than polymers, e.g. ceramic silicates, are currently being investigated for gas separation processes. The permeation characteristics of one such material, Vycor (Corning Glass #1370), have been reported for the separation of hydrogen from hydrogen iodide. This paper will describe the electron microscopy techniques applied to reveal the porous microstructure of a Vycor membrane. The application of these techniques has led to an increased understanding in the relationship between the substructure and the gas transport properties of this material.


Author(s):  
T. Geipel ◽  
W. Mader ◽  
P. Pirouz

Temperature affects both elastic and inelastic scattering of electrons in a crystal. The Debye-Waller factor, B, describes the influence of temperature on the elastic scattering of electrons, whereas the imaginary part of the (complex) atomic form factor, fc = fr + ifi, describes the influence of temperature on the inelastic scattering of electrons (i.e. absorption). In HRTEM simulations, two possible ways to include absorption are: (i) an approximate method in which absorption is described by a phenomenological constant, μ, i.e. fi; - μfr, with the real part of the atomic form factor, fr, obtained from Hartree-Fock calculations, (ii) a more accurate method in which the absorptive components, fi of the atomic form factor are explicitly calculated. In this contribution, the inclusion of both the Debye-Waller factor and absorption on HRTEM images of a (Oll)-oriented GaAs crystal are presented (using the EMS software.Fig. 1 shows the the amplitudes and phases of the dominant 111 beams as a function of the specimen thickness, t, for the cases when μ = 0 (i.e. no absorption, solid line) and μ = 0.1 (with absorption, dashed line).


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