scholarly journals Quantitative mapping of triacylglycerol chain length and saturation using broadband CARS microscopy

2018 ◽  
Author(s):  
A. Paul ◽  
Y.J. Wang ◽  
C. Brännmark ◽  
S. Kumar ◽  
M. Bonn ◽  
...  
Keyword(s):  
Chain Length ◽  
High Speed ◽  
Lipid Analysis ◽  
Neutral Lipids ◽  
Cell Types ◽  
Label Free ◽  
Complex Samples ◽  
Disease States ◽  
Coherent Raman

AbstractLipid droplets (LDs) are highly dynamic organelles that store neutral lipids, primarily triacylglycerols (TAGs), and are found in many cell types. While their primary function is to store excess energy, LDs are also modified in different disease states and during developmental processes. In many cases, not only the presence, but also the composition, of LDs can be equally important. In humans, LD composition has been linked to diseases such as type 2 diabetes; in plants and yeast, LD composition is relevant for engineering these organisms into biological factories in, e.g., algal bioenergy or food oil production. Therefore, lipid analysis of biological LDs with high speed and accuracy in situ is a very active area of research. Here we present an approach for in situ, quantitative TAG analysis using label-free, coherent Raman microscopy to decipher LD TAG composition in different biochemically complex samples. Our method allows direct visualization of inter-LD compositional heterogeneity of physiological quantities – TAG chain length and number of C=C bonds – with sub-micrometer spatial resolution within 5-100 milliseconds. Combined with virtually no sample preparation, this approach should enable rapid and accurate TAG LD analysis for a variety of applications.

Mucin mRNA Expression in Normal and Vasomotor Inferior Turbinates

1997 ◽  
Vol 11 (4) ◽  
pp. 293-302 ◽  
Author(s):  
Michelle R. Aust ◽  
Cathy S. Madsen ◽  
Anita Jennings ◽  
Jan L. Kasperbauer ◽  
Sandra J. Gendler
Keyword(s):  
Respiratory Tract ◽  
Expression Patterns ◽  
Inferior Turbinate ◽  
Cell Types ◽  
Upper Respiratory Tract ◽  
Vasomotor Rhinitis ◽  
Disease States ◽  
Multiple Cell ◽  
Nasal Secretions

Mucins are the major glycoprotein component of respiratory tract secretions. Little is known about their expression in the upper respiratory tract. In order to define this expression, in situ hybridization was performed on 19 normal and 4 vasomotor rhinitis (VMR) inferior turbinates to identify mucin mRNA. MUC1, MUC2, MUC4, MUC5AC, MUC5B, and MUC7 were expressed in both the normal and VMR turbinates. MUC 4 and MUC5AC were the most highly expressed mucins. MUC1, MUC2, MUC4, and MUC5AC were expressed mainly by the epithelial border, whereas MUC5B and MUC7 were expressed by the submucosal glands. MUC1 and MUC4 exhibited a diffuse expression by multiple cell types along the mucosal border, whereas MUC2 and MUC5AC expression appeared to be limited to a subpopulation of epithelial cells, most likely goblet cells. Although MUC1, MUC4, and MUC5AC showed sporadic submucosal glandular expression, MUC5B and MUC7 appeared to be the predominant submucosal gland mucins in the inferior turbinates. MUC3 and MUC6 expression, which have been found primarily in the gastric mucosa, were not seen in any of the inferior turbinate samples examined. The only difference seen between normal and VMR turbinates was a slight decrease in MUC1 expression in the VMR group. The variety of mucins expressed and the diversity of their expression patterns may have significance in terms of the rheologic and particle clearance properties of nasal secretions. Understanding the expression patterns in normal turbinates will serve as the foundation for further study of these mucins in disease states.

scholarly journals Label-free in situ imaging of oil body dynamics and chemistry in germination

2016 ◽  
Vol 13 (123) ◽  
pp. 20160677 ◽  
Author(s):  
Gustav Waschatko ◽  
Nils Billecke ◽  
Sascha Schwendy ◽  
Henriette Jaurich ◽  
Mischa Bonn ◽  
...  
Keyword(s):  
Size Composition ◽  
Primary Energy ◽  
Oil Body ◽  
Label Free ◽  
Oil Bodies ◽  
Lipid Dynamics ◽  
Body Dynamics ◽  
Body Location ◽  
Coherent Raman

Plant oleosomes are uniquely emulsified lipid reservoirs that serve as the primary energy source during seed germination. These oil bodies undergo significant changes regarding their size, composition and structure during normal seedling development; however, a detailed characterization of these oil body dynamics, which critically affect oil body extractability and nutritional value, has remained challenging because of a limited ability to monitor oil body location and composition during germination in situ . Here, we demonstrate via in situ , label-free imaging that oil bodies are highly dynamic intracellular organelles that are morphologically and biochemically remodelled extensively during germination. Label-free, coherent Raman microscopy (CRM) combined with bulk biochemical measurements revealed the temporal and spatial regulation of oil bodies in native soya bean cotyledons during the first eight days of germination. Oil bodies undergo a cycle of growth and shrinkage that is paralleled by lipid and protein compositional changes. Specifically, the total protein concentration associated with oil bodies increases in the first phase of germination and subsequently decreases. Lipids contained within the oil bodies change in saturation and chain length during germination. Our results show that CRM is a well-suited platform to monitor in situ lipid dynamics and local chemistry and that oil bodies are actively remodelled during germination. This underscores the dynamic role of lipid reservoirs in plant development.

scholarly journals Label-free CARS microscopy reveals similar triacylglycerol acyl chain length and saturation in myocellular lipid droplets of athletes and individuals with type 2 diabetes

Diabetologia ◽  
2020 ◽  
Vol 63 (12) ◽  
pp. 2654-2664
Author(s):  
Sabine Daemen ◽  
Anne Gemmink ◽  
Alexandra Paul ◽  
Nils Billecke ◽  
Katrina Rieger ◽  
...  
Keyword(s):  
Type 2 Diabetes ◽  
Insulin Sensitivity ◽  
Chain Length ◽  
Acyl Chain ◽  
Subcellular Location ◽  
Label Free ◽  
Acyl Chain Length ◽  
Type 2 Diabetic

Abstract Aims/hypothesis Intramyocellular lipid (IMCL) content associates with development of insulin resistance, albeit not in insulin-sensitive endurance-trained athletes (trained). Qualitative and spatial differences in muscle lipid composition may underlie this so-called athlete’s paradox. Here we studied triacylglycerol (TAG) composition of individual myocellular lipid droplets (LDs) in trained individuals and individuals with type 2 diabetes mellitus. Methods Trained ($$ \dot{V}{\mathrm{O}}_{2\max } $$ V ̇ O 2 max 71.0 ± 1.6 ml O2 [kg lean body mass (LBM)]−1 min−1), normoglycaemic (fasting glucose 5.1 ± 0.1 mmol/l) individuals and untrained ($$ \dot{V}{\mathrm{O}}_{2\max } $$ V ̇ O 2 max 36.8 ± 1.5 ml O2 [kg LBM]−1 min−1) individuals with type 2 diabetes (fasting glucose 7.4 ± 0.5 mmol/l), with similar IMCL content (3.5 ± 0.7% vs 2.5 ± 0.3%, p = 0.241), but at opposite ends of the insulin sensitivity spectrum (glucose infusion rate 93.8 ± 6.6 vs 25.7 ± 5.3 μmol [kg LBM]−1 min−1 for trained individuals and those with type 2 diabetes, respectively) were included from our database in the present study. We applied in situ label-free broadband coherent anti-Stokes Raman scattering (CARS) microscopy to sections from skeletal muscle biopsies to measure TAG acyl chain length and saturation of myocellular LDs. This approach uniquely permits examination of individual LDs in their native environment, in a fibre-type-specific manner, taking into account LD size and subcellular location. Results Despite a significant difference in insulin sensitivity, we observed remarkably similar acyl chain length and saturation in trained and type 2 diabetic individuals (chain length: 18.12 ± 0.61 vs 18.36 ± 0.43 number of carbons; saturation: 0.37 ± 0.05 vs 0.38 ± 0.06 number of C=C bonds). Longer acyl chains or higher saturation (lower C=C number) could be detected in subpopulations of LDs, i.e. large LDs (chain length: 18.11 ± 0.48 vs 18.63 ± 0.57 carbon number) and subsarcolemmal LDs (saturation: 0.34 ± 0.02 vs 0.36 ± 0.04 C=C number), which are more abundant in individuals with type 2 diabetes. Conclusions/interpretation In contrast to reports of profound differences in the lipid composition of lipids extracted from skeletal muscle from trained and type 2 diabetic individuals, our in situ, LD-specific approach detected only modest differences in TAG composition in LD subpopulations, which were dependent on LD size and subcellular location. If, and to what extent, these modest differences can impact insulin sensitivity remains to be elucidated.

scholarly journals Temporal Imaging of Live Cells by High-Speed Confocal Raman Microscopy

Materials ◽  
2021 ◽  
Vol 14 (13) ◽  
pp. 3732
Author(s):  
Jeon Woong Kang ◽  
Freddy T. Nguyen ◽  
Niyom Lue
Keyword(s):  
High Speed ◽  
External Perturbation ◽  
Cell Types ◽  
Raman Microscopy ◽  
Live Cells ◽  
Confocal Raman Microscopy ◽  
Label Free ◽  
Minimal Sample ◽  
Confocal Raman

Label-free live cell imaging was performed using a custom-built high-speed confocal Raman microscopy system. For various cell types, cell-intrinsic Raman bands were monitored. The high-resolution temporal Raman images clearly delineated the intracellular distribution of biologically important molecules such as protein, lipid, and DNA. Furthermore, optical phase delay measured using quantitative phase microscopy shows similarity with the image reconstructed from the protein Raman peak. This reported work demonstrates that Raman imaging is a powerful label-free technique for studying various biomedical problems in vitro with minimal sample preparation and external perturbation to the cellular system.

An atomic-resolution microscope study of Al contracts on GaAs

1986 ◽  
Vol 44 ◽  
pp. 726-727
Author(s):  
Z. Liliental-Weber ◽  
C. Nelson ◽  
R. Ludeke ◽  
R. Gronsky ◽  
J. Washburn
Keyword(s):  
High Speed ◽  
Schottky Contacts ◽  
Detailed Knowledge ◽  
The Past ◽  
Semiconductor Interfaces ◽  
Deposited Metal ◽  
Microwave Applications ◽  
Free Interfaces ◽  
Potential Use

The properties of metal/semiconductor interfaces have received considerable attention over the past few years, and the Al/GaAs system is of special interest because of its potential use in high-speed logic integrated optics, and microwave applications. For such materials a detailed knowledge of the geometric and electronic structure of the interface is fundamental to an understanding of the electrical properties of the contact. It is well known that the properties of Schottky contacts are established within a few atomic layers of the deposited metal. Therefore surface contamination can play a significant role. A method for fabricating contamination-free interfaces is absolutely necessary for reproducible properties, and molecularbeam epitaxy (MBE) offers such advantages for in-situ metal deposition under UHV conditions

scholarly journals Spatially Resolved Experimental Modal Analysis on High-Speed Composite Rotors Using a Non-Contact, Non-Rotating Sensor

Sensors ◽  
2021 ◽  
Vol 21 (14) ◽  
pp. 4705
Author(s):  
Julian Lich ◽  
Tino Wollmann ◽  
Angelos Filippatos ◽  
Maik Gude ◽  
Juergen Czarske ◽  
...  
Keyword(s):  
High Speed ◽  
Natural Frequencies ◽  
Mode Shapes ◽  
Fiber Reinforced ◽  
Structural Dynamic ◽  
Spatially Resolved ◽  
Glass Fiber Reinforced ◽  
Vibration Responses ◽  
And Performance

Due to their lightweight properties, fiber-reinforced composites are well suited for large and fast rotating structures, such as fan blades in turbomachines. To investigate rotor safety and performance, in situ measurements of the structural dynamic behaviour must be performed during rotating conditions. An approach to measuring spatially resolved vibration responses of a rotating structure with a non-contact, non-rotating sensor is investigated here. The resulting spectra can be assigned to specific locations on the structure and have similar properties to the spectra measured with co-rotating sensors, such as strain gauges. The sampling frequency is increased by performing consecutive measurements with a constant excitation function and varying time delays. The method allows for a paradigm shift to unambiguous identification of natural frequencies and mode shapes with arbitrary rotor shapes and excitation functions without the need for co-rotating sensors. Deflection measurements on a glass fiber-reinforced polymer disk were performed with a diffraction grating-based sensor system at 40 measurement points with an uncertainty below 15 μrad and a commercial triangulation sensor at 200 measurement points at surface speeds up to 300 m/s. A rotation-induced increase of two natural frequencies was measured, and their mode shapes were derived at the corresponding rotational speeds. A strain gauge was used for validation.

scholarly journals In situ differentiation of iridophore crystallotypes underlies zebrafish stripe patterning

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Dvir Gur ◽  
Emily J. Bain ◽  
Kory R. Johnson ◽  
Andy J. Aman ◽  
H. Amalia Pasoili ◽  
...  
Keyword(s):  
Skin Color ◽  
Cell Types ◽  
Color Pattern ◽  
Single Type ◽  
Sequencing Analysis ◽  
X Ray Diffraction ◽  
X Ray ◽  
Cryogenic Scanning Electron Microscopy ◽  
Different Cell Types

AbstractSkin color patterns are ubiquitous in nature, impact social behavior, predator avoidance, and protection from ultraviolet irradiation. A leading model system for vertebrate skin patterning is the zebrafish; its alternating blue stripes and yellow interstripes depend on light-reflecting cells called iridophores. It was suggested that the zebrafish’s color pattern arises from a single type of iridophore migrating differentially to stripes and interstripes. However, here we find that iridophores do not migrate between stripes and interstripes but instead differentiate and proliferate in-place, based on their micro-environment. RNA-sequencing analysis further reveals that stripe and interstripe iridophores have different transcriptomic states, while cryogenic-scanning-electron-microscopy and micro-X-ray diffraction identify different crystal-arrays architectures, indicating that stripe and interstripe iridophores are different cell types. Based on these results, we present an alternative model of skin patterning in zebrafish in which distinct iridophore crystallotypes containing specialized, physiologically responsive, organelles arise in stripe and interstripe by in-situ differentiation.

Enhanced Grain Refinement of W18Cr4V High‐Speed Steel using in situ TiN‐Nb‐Cr@Graphene/Fe Nanocomposite Inoculant

2021 ◽  
Author(s):  
Lina Bai ◽  
Chunxiang Cui ◽  
Jianjun Zhang ◽  
Lichen Zhao ◽  
Guixing Zheng ◽  
...  
Keyword(s):  
Grain Refinement ◽  
High Speed ◽  
High Speed Steel

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.

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