scholarly journals Circular Intensity Differential Scattering for Label-Free Chromatin Characterization: A Review for Optical Microscopy

Polymers ◽  
2020 ◽  
Vol 12 (10) ◽  
pp. 2428
Author(s):  
Aymeric Le Gratiet ◽  
Riccardo Marongiu ◽  
Alberto Diaspro
Keyword(s):  
Optical Microscopy ◽  
Gold Standard ◽  
Molecular Conformation ◽  
Polarized Light ◽  
Excitation Wavelength ◽  
General Description ◽  
Label Free ◽  
Differential Measurement ◽  
Scale Down

Circular Intensity Differential Scattering (CIDS) provides a differential measurement of the circular right and left polarized light and has been proven to be a gold standard label-free technique to study the molecular conformation of complex biopolymers, such as chromatin. In early works, it has been shown that the scattering component of the CIDS signal gives information from the long-range chiral organization on a scale down to 1/10th–1/20th of the excitation wavelength, leading to information related to the structure and orientation of biopolymers in situ at the nanoscale. In this paper, we review the typical methods and technologies employed for measuring this signal coming from complex macro-molecules ordering. Additionally, we include a general description of the experimental architectures employed for spectroscopic CIDS measurements, angular or spectral, and of the most recent advances in the field of optical imaging microscopy, allowing a visualization of the chromatin organization in situ.

Label-free fluorescent discrimination and detection of epinephrine and dopamine based on bioinspired in situ copolymers and excitation wavelength switch

2019 ◽  
Vol 1054 ◽  
pp. 167-175 ◽  
Author(s):  
Ying Zhang ◽  
Wang Ren ◽  
Yu Zhu Fan ◽  
Jiang Xue Dong ◽  
Hui Zhang ◽  
...  
Keyword(s):  
Excitation Wavelength ◽  
Label Free

scholarly journals Multi-pronged approach to human mesenchymal stromal cells senescence quantification with a focus on label-free methods

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Weichao Zhai ◽  
Jerome Tan ◽  
Tobias Russell ◽  
Sixun Chen ◽  
Dennis McGonagle ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Preclinical Model ◽  
Label Free ◽  
Current Standard ◽  
Differentiation Potential ◽  
Human Mesenchymal Stromal Cells ◽  
Endogenous Fluorophores

AbstractHuman mesenchymal stromal cells (hMSCs) have demonstrated, in various preclinical settings, consistent ability in promoting tissue healing and improving outcomes in animal disease models. However, translation from the preclinical model into clinical practice has proven to be considerably more difficult. One key challenge being the inability to perform in situ assessment of the hMSCs in continuous culture, where the accumulation of the senescent cells impairs the culture’s viability, differentiation potential and ultimately leads to reduced therapeutic efficacies. Histochemical $$\upbeta $$ β -galactosidase staining is the current standard for measuring hMSC senescence, but this method is destructive and not label-free. In this study, we have investigated alternatives in quantification of hMSCs senescence, which included flow cytometry methods that are based on a combination of cell size measurements and fluorescence detection of SA-$$\upbeta $$ β -galactosidase activity using the fluorogenic substrate, C$${_{12}}$$ 12 FDG; and autofluorescence methods that measure fluorescence output from endogenous fluorophores including lipopigments. For identification of senescent cells in the hMSC batches produced, the non-destructive and label-free methods could be a better way forward as they involve minimum manipulations of the cells of interest, increasing the final output of the therapeutic-grade hMSC cultures. In this work, we have grown hMSC cultures over a period of 7 months and compared early and senescent hMSC passages using the advanced flow cytometry and autofluorescence methods, which were benchmarked with the current standard in $$\upbeta $$ β -galactosidase staining. Both the advanced methods demonstrated statistically significant values, (r = 0.76, p $$\le $$ ≤ 0.001 for the fluorogenic C$${_{12}}$$ 12 FDG method, and r = 0.72, p $$\le $$ ≤ 0.05 for the forward scatter method), and good fold difference ranges (1.120–4.436 for total autofluorescence mean and 1.082–6.362 for lipopigment autofluorescence mean) between early and senescent passage hMSCs. Our autofluroescence imaging and spectra decomposition platform offers additional benefit in label-free characterisation of senescent hMSC cells and could be further developed for adoption for future in situ cellular senescence evaluation by the cell manufacturers.

scholarly journals Label-Free and Quantitative Dry Mass Monitoring for Single Cells during In Situ Culture

Cells ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1635
Author(s):  
Ya Su ◽  
Rongxin Fu ◽  
Wenli Du ◽  
Han Yang ◽  
Li Ma ◽  
...  
Keyword(s):  
Cell Growth ◽  
Single Cell ◽  
Hela Cells ◽  
Quantitative Measurement ◽  
Single Cells ◽  
Dry Mass ◽  
Quantitative Detection ◽  
Label Free ◽  
Mass Sensitivity

Quantitative measurement of single cells can provide in-depth information about cell morphology and metabolism. However, current live-cell imaging techniques have a lack of quantitative detection ability. Herein, we proposed a label-free and quantitative multichannel wide-field interferometric imaging (MWII) technique with femtogram dry mass sensitivity to monitor single-cell metabolism long-term in situ culture. We demonstrated that MWII could reveal the intrinsic status of cells despite fluctuating culture conditions with 3.48 nm optical path difference sensitivity, 0.97 fg dry mass sensitivity and 2.4% average maximum relative change (maximum change/average) in dry mass. Utilizing the MWII system, different intrinsic cell growth characteristics of dry mass between HeLa cells and Human Cervical Epithelial Cells (HCerEpiC) were studied. The dry mass of HeLa cells consistently increased before the M phase, whereas that of HCerEpiC increased and then decreased. The maximum growth rate of HeLa cells was 11.7% higher than that of HCerEpiC. Furthermore, HeLa cells were treated with Gemcitabine to reveal the relationship between single-cell heterogeneity and chemotherapeutic efficacy. The results show that cells with higher nuclear dry mass and nuclear density standard deviations were more likely to survive the chemotherapy. In conclusion, MWII was presented as a technique for single-cell dry mass quantitative measurement, which had significant potential applications for cell growth dynamics research, cell subtype analysis, cell health characterization, medication guidance and adjuvant drug development.

scholarly journals In situ surface-enhanced Raman spectroelectrochemistry reveals the molecular conformation of electrolyte additives in Li-ion batteries

2021 ◽  
Author(s):  
Chenbo Zhu ◽  
Chenghao Fan ◽  
Emiliano Cortes ◽  
Wei Xie
Keyword(s):  
Molecular Conformation ◽  
Rate Capability ◽  
Cycle Performance ◽  
Li Ion Batteries ◽  
Electrolyte Additive ◽  
Electrolyte Additives ◽  
Surface Enhanced ◽  
Surface Enhanced Raman ◽  
Li Ion

We report on the mechanism of rhodamine B (RhB) acting as an electrolyte additive in Li/graphite cells. We show that cycle performance and rate capability of graphite is enhanced in...

In Situ Magnetic-Circular-X-Ray-Dichroism Measurements: An Epitaxial Fe Wedge on Cu(100)

MRS Bulletin ◽  
1999 ◽  
Vol 24 (1) ◽  
pp. 41-45 ◽  
Author(s):  
M.E. Dávila ◽  
D. Arvanitis ◽  
J. Hunter Dunn ◽  
N. Mårtensson ◽  
P. Srivastava ◽  
...  
Keyword(s):  
Ultrathin Films ◽  
Magnetic Circular Dichroism ◽  
Polarized Light ◽  
Measured Quantity ◽  
Easy Magnetization ◽  
Magnetization Direction ◽  
Core Electron ◽  
Circularly Polarized ◽  
X Ray

Circularly polarized x-ray radiation is attracting increasing interest as a tool for the characterization of the electronic, magnetic, and chiral properties of low-dimensional structures. Using circular light (with electric field vector parallel to the orbital plane), a dependence of the measured quantity by changing either the orientation of the light polarization or the magnetization is indicative of the existence of magnetic circular dichroism. It can be observed in x-ray absorption spectroscopy (XAS), in which the photon energy is scanned through an absorption threshold exciting a core electron into an unoccupied valence state using circularly polarized light. Synchrotron radiation sources have made this technique possible. It can also be observed in photo-emission spectroscopy from core and valence levels. Here we focus on magnetic circular x-ray dichroism (MCXD) in XAS as an element-specific tool to investigate magnetic properties of ultrathin films in situ. The application of magneto-optical sum rules enables the determination of the orbital and spin magnetic moments per atom from XAS spectra, as well as the easy magnetization direction.MCXD-based magnetometry in XAS is extensively used by measuring the L absorption edges of 3d-transition metals, where large intensity changes (up to 60%) of the L-edge white lines are observed upon reversal of either the sample magnetization or the light helicity. The high magnetic contrast obtained, combined with the elemental specificity of the technique, allows for the study of very dilute samples such as ultrathin films. We first concentrate on the selection rules governing MCXD in XAS.

scholarly journals In-situ SEM and optical microscopy testing for investigation of fatigue crack growth mechanism under overload

2018 ◽  
Vol 165 ◽  
pp. 13013
Author(s):  
Wei Zhang ◽  
Liang Cai
Keyword(s):  
Growth Rate ◽  
Crack Growth Rate ◽  
Optical Microscopy ◽  
Crack Growth ◽  
Load Condition ◽  
Crack Tip ◽  
Initial Crack ◽  
Crack Growth Behavior ◽  
Single Overload

In this paper, the in-situ scanning electron microscope (SEM) and optical microscopy experiments are performed to investigate the crack growth behavior under the single tensile overload. The objectives are to (i) examine the overload-induced crack growth micromechanisms, including the initial crack growth acceleration and the subsequent retardation period; (ii) investigate the effective region of single overload on crack growth rate. The specimen is a small thin Al2024-T3 plate with an edge-crack, which is loaded and observed in the SEM chamber. The very high resolution images of the crack tip are taken under the simple variable amplitude loading. Imaging analysis is performed to quantify the crack tip deformation at any time instant. Moreover, an identical specimen subjected to the same load condition is observed under optical microscope. In this testing, fine speckling is performed to promote the accuracy of digital imaging correlation (DIC). The images around the crack tip are taken at the peak loads before, during and after the single overload. After that, the evolution of local strain distribution is obtained through DIC technique. The results show that the rapid connection between the main crack and microcracks accounts for the initial crack growth acceleration. The crack closure level can be responsible for the crack growth rate during the steady growth period. Besides that, the size of retardation area is larger than the classical solution.

The Effect of Stand-Off Distance on the Structure and Properties of Zirconium – Carbon Steel Bimetal Produced by Explosion Welding / Wpływ Odległości Blach Na Strukturę I Własności Bimetalu Cyrkon - Stal Wytworzonego Technologią Zgrzewania Wybuchowego

2012 ◽  
Vol 57 (4) ◽  
pp. 1201-1210 ◽  
Author(s):  
M. Prażmowski ◽  
H. Paul
Keyword(s):  
Carbon Steel ◽  
Optical Microscopy ◽  
Longitudinal Section ◽  
Strength Properties ◽  
Strength Distribution ◽  
Explosion Welding ◽  
General Description ◽  
Bond Zone ◽  
Bond Interface ◽  
The Individual

This study focuses on the effect of the stand-off distance between the bonded plates on the properties of zirconium (Zr700) - steel (P355NL2) bimetal produced by explosion welding. Bonding trials were carried out in parallel arrangement at constant detonation velocity. The analyses of microstructural transformations occurring in the bond zone and mechanical properties of the clad were performed for as-bonded welds, i.e. immediately following explosion welding. A general description of the obtained welds was made (height and length of the wave was determined) and the quantitative fraction of the melt zones was calculated along the bond’s length. Using optical microscopy and scanning electron microscopy (SEM) enabled the assessment of the quality of the formed bonds, initial identification of phases and quantitative analysis of the individual phases on the longitudinal section. The microhardness results were used in the analysis of hardening changes at the interface area. The completed research proves the potential to obtain a proper bond for zirconium/carbon steel sheets. A strong effect of the stand-off distance on the strength properties of the fabricated plates was observed, and the ’direction’ of these transformations was pointed out. Optical microscopy and SEM examinations allowed determining the characteristic of the bond interface for diverse stand-off distances. It was established that increasing the stand-off distance between the plates causes the reduction of the melt area along the length of the bond, which improves strength properties of the bimetal. The analysis of the strength distribution performed based on the microhardness measurements showed that the changes occur within the distances up to 500µm from the bond interface and the highest hardening, for both zirconium and steel, is directly at the interface and then successively decreases.

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