scholarly journals Quantitative Phase Velocimetry for Label-Free Measurement of Intracellular Mass Transport Velocity

2021 ◽  
Author(s):  
Soorya Pradeep ◽  
Thomas A Zangle
Keyword(s):  
Synthetic Data ◽  
Label Free ◽  
Quantitative Phase ◽  
Cell Compartments ◽  
Distinct Cell ◽  
Effective Particle ◽  
Mass Transport Velocity ◽  
Entire Flow ◽  
Single Approach

Transport of mass within cells helps maintain homeostasis and is disrupted by disease and stress. Here, we develop quantitative phase velocimetry (QPV) as a label-free approach to make the invisible flow of mass within cells visible and quantifiable. We benchmark our approach against alternative image registration methods, a theoretical error model, and synthetic data. Our method tracks not just individual labeled particles or molecules, but the entire flow of material through the cell. This enables us to measure diffusivity within distinct cell compartments using a single approach, which we use here for direct comparison of nuclear and cytoplasmic diffusivity. As a label-free method, QPV can be used for long-term tracking to capture dynamics through the cell cycle. Finally, based on the known effective particle size, we show that QPV is an accessible method to measure intracellular viscosity.

scholarly journals Characterization of Liposomes Using Quantitative Phase Microscopy (QPM)

Pharmaceutics ◽  
2021 ◽  
Vol 13 (5) ◽  
pp. 590
Author(s):  
Jennifer Cauzzo ◽  
Nikhil Jayakumar ◽  
Balpreet Singh Ahluwalia ◽  
Azeem Ahmad ◽  
Nataša Škalko-Basnet
Keyword(s):  
Rapid Development ◽  
Fluorescent Labeling ◽  
Label Free ◽  
Batch Mode ◽  
Full Field ◽  
Quantitative Phase ◽  
Phase Microscopy ◽  
Quantitative Phase Microscopy ◽  
Tracking Ability

The rapid development of nanomedicine and drug delivery systems calls for new and effective characterization techniques that can accurately characterize both the properties and the behavior of nanosystems. Standard methods such as dynamic light scattering (DLS) and fluorescent-based assays present challenges in terms of system’s instability, machine sensitivity, and loss of tracking ability, among others. In this study, we explore some of the downsides of batch-mode analyses and fluorescent labeling, while introducing quantitative phase microscopy (QPM) as a label-free complimentary characterization technique. Liposomes were used as a model nanocarrier for their therapeutic relevance and structural versatility. A successful immobilization of liposomes in a non-dried setup allowed for static imaging conditions in an off-axis phase microscope. Image reconstruction was then performed with a phase-shifting algorithm providing high spatial resolution. Our results show the potential of QPM to localize subdiffraction-limited liposomes, estimate their size, and track their integrity over time. Moreover, QPM full-field-of-view images enable the estimation of a single-particle-based size distribution, providing an alternative to the batch mode approach. QPM thus overcomes some of the drawbacks of the conventional methods, serving as a relevant complimentary technique in the characterization of nanosystems.

scholarly journals Nanoparticle Determination in Water by LED-Excited Surface Plasmon Resonance Imaging

Chemosensors ◽  
2021 ◽  
Vol 9 (7) ◽  
pp. 175
Author(s):  
Lukas Wunderlich ◽  
Peter Hausler ◽  
Susanne Märkl ◽  
Rudolf Bierl ◽  
Thomas Hirsch
Keyword(s):  
Light Source ◽  
Self Assembled Monolayers ◽  
Receptor Interaction ◽  
Complex Nature ◽  
Label Free ◽  
Quality Of Water ◽  
Spr Imaging ◽  
Online Sensor ◽  
Assembled Monolayers

The increasing popularity of nanoparticles in many applications has led to the fact that these persistent materials pollute our environment and threaten our health. An online sensor system for monitoring the presence of nanoparticles in fresh water would be highly desired. We propose a label-free sensor based on SPR imaging. The sensitivity was enhanced by a factor of about 100 by improving the detector by using a high-resolution camera. This revealed that the light source also needed to be improved by using LED excitation instead of a laser light source. As a receptor, different self-assembled monolayers have been screened. It can be seen that the nanoparticle receptor interaction is of a complex nature. The best system when taking sensitivity as well as reversibility into account is given by a dodecanethiol monolayer on the gold sensor surface. Lanthanide-doped nanoparticles, 29 nm in diameter and with a similar refractive index to the most common silica nanoparticles were detected in water down to 1.5 µg mL−1. The sensor can be fully regenerated within one hour without the need for any washing buffer. This sensing concept is expected to be easily adapted for the detection of nanoparticles of different size, shape, and composition, and upon miniaturization, suitable for long-term applications to monitor the quality of water.

scholarly journals Analysis of full disc Ca II K spectroheliograms

2018 ◽  
Vol 609 ◽  
pp. A92 ◽  
Author(s):  
Theodosios Chatzistergos ◽  
Ilaria Ermolli ◽  
Sami K. Solanki ◽  
Natalie A. Krivova
Keyword(s):  
Synthetic Data ◽  
Calibration Method ◽  
Poor Quality ◽  
Average Error ◽  
Photometric Calibration ◽  
Solar Observations ◽  
Relative Errors ◽  
Long Term Variations ◽  
The Ideal

Context. Historical Ca II K spectroheliograms (SHG) are unique in representing long-term variations of the solar chromospheric magnetic field. They usually suffer from numerous problems and lack photometric calibration. Thus accurate processing of these data is required to get meaningful results from their analysis. Aims. In this paper we aim at developing an automatic processing and photometric calibration method that provides precise and consistent results when applied to historical SHG. Methods. The proposed method is based on the assumption that the centre-to-limb variation of the intensity in quiet Sun regions does not vary with time. We tested the accuracy of the proposed method on various sets of synthetic images that mimic problems encountered in historical observations. We also tested our approach on a large sample of images randomly extracted from seven different SHG archives. Results. The tests carried out on the synthetic data show that the maximum relative errors of the method are generally <6.5%, while the average error is <1%, even if rather poor quality observations are considered. In the absence of strong artefacts the method returns images that differ from the ideal ones by <2% in any pixel. The method gives consistent values for both plage and network areas. We also show that our method returns consistent results for images from different SHG archives. Conclusions. Our tests show that the proposed method is more accurate than other methods presented in the literature. Our method can also be applied to process images from photographic archives of solar observations at other wavelengths than Ca II K.

scholarly journals Label-Free Study of the Global Cell Behavior during Exposure to Environmental Radiofrequency Fields in the Presence or Absence of Pro-Apoptotic or Pro-Autophagic Treatments

2022 ◽  
Vol 23 (2) ◽  
pp. 658
Author(s):  
Alexandre Joushomme ◽  
André Garenne ◽  
Mélody Dufossée ◽  
Rémy Renom ◽  
Hermanus Johannes Ruigrok ◽  
...  
Keyword(s):  
Mobile Communications ◽  
Cellular Response ◽  
Continuous Wave ◽  
Digital Holographic Microscopy ◽  
Label Free ◽  
Environmental Signals ◽  
Chemical Treatments ◽  
Rf Exposure ◽  
Universal Mobile Telecommunications System

It remains controversial whether exposure to environmental radiofrequency signals (RF) impacts cell status or response to cellular stress such as apoptosis or autophagy. We used two label-free techniques, cellular impedancemetry and Digital Holographic Microscopy (DHM), to assess the overall cellular response during RF exposure alone, or during co-exposure to RF and chemical treatments known to induce either apoptosis or autophagy. Two human cell lines (SH-SY5Y and HCT116) and two cultures of primary rat cortex cells (astrocytes and co-culture of neurons and glial cells) were exposed to RF using an 1800 MHz carrier wave modulated with various environmental signals (GSM: Global System for Mobile Communications, 2G signal), UMTS (Universal Mobile Telecommunications System, 3G signal), LTE (Long-Term Evolution, 4G signal, and Wi-Fi) or unmodulated RF (continuous wave, CW). The specific absorption rates (S.A.R.) used were 1.5 and 6 W/kg during DHM experiments and ranged from 5 to 24 W/kg during the recording of cellular impedance. Cells were continuously exposed for three to five consecutive days while the temporal phenotypic signature of cells behavior was recorded at constant temperature. Statistical analysis of the results does not indicate that RF-EMF exposure impacted the global behavior of healthy, apoptotic, or autophagic cells, even at S.A.R. levels higher than the guidelines, provided that the temperature was kept constant.

scholarly journals The phosphoproteome of rice leaves responds to water and nitrogen supply

2021 ◽  
Author(s):  
Sara Hamzelou ◽  
Vanessa J. Melino ◽  
Darren C. Plett ◽  
Karthik Shantharam Kamath ◽  
Arkadiusz Nawrocki ◽  
...  
Keyword(s):  
Prolonged Exposure ◽  
Water Channel ◽  
Nitrogen Supply ◽  
Nitrogen Fertilizers ◽  
Label Free ◽  
Limited Information ◽  
Irrigated Rice ◽  
Post Translational Modifications ◽  
Growth Adaptation

The scarcity of freshwater is an increasing concern in flood-irrigated rice, whilst excessive use of nitrogen fertilizers is both costly and contributes to environmental pollution. To co-ordinate growth adaptation under prolonged exposure to limited water or excess nitrogen supply, plants have processes for signalling and regulation of metabolic processes. There is limited information on the involvement of one of the most important post-translational modifications (PTMs), protein phosphorylation, on plant adaptation to long-term changes in resource supply. Oryza sativa cv. Nipponbare was grown under two regimes of nitrogen from the time of germination to final harvest. Twenty-five days after germination, water was withheld from half the pots in each nitrogen treatment and low water supply continued for an additional 26 days, while the remaining pots were well watered. Leaves from all four groups of plants were harvested after 51 days in order to test whether phosphorylation of leaf proteins responded to prior abiotic events. The dominant impact of these resources is exerted in leaves, where PTMs have been predicted to occur. Proteins were extracted and phosphopeptides were analysed by nanoLC-MS/MS analysis, coupled with label-free quantitation. Water and nitrogen regimes triggered extensive changes in phosphorylation of proteins involved in membrane transport, such as the aquaporin OsPIP2-6, a water channel protein. Our study reveals phosphorylation of several peptides belonging to proteins involved in RNA-processing and carbohydrate metabolism, suggesting that phosphorylation events regulate the signalling cascades that are required to optimize plant response to resource supply.

scholarly journals Estimating activity cycles with probabilistic methods

2018 ◽  
Vol 615 ◽  
pp. A111 ◽  
Author(s):  
N. Olspert ◽  
J. Pelt ◽  
M. J. Käpylä ◽  
J. Lehtinen
Keyword(s):  
Linear Trend ◽  
Synthetic Data ◽  
Probabilistic Methods ◽  
Data Sampling ◽  
Constant Variance ◽  
Noise Type ◽  
Periodic Model ◽  
Activity Cycles ◽  
Magnetic Cycles

Context. Period estimation is one of the central topics in astronomical time series analysis, in which data is often unevenly sampled. Studies of stellar magnetic cycles are especially challenging, as the periods expected in those cases are approximately the same length as the datasets themselves. The datasets often contain trends, the origin of which is either a real long-term cycle or an instrumental effect. But these effects cannot be reliably separated, while they can lead to erroneous period determinations if not properly handled. Aims. In this study we aim at developing a method that can handle the trends properly. By performing an extensive set of testing, we show that this is the optimal procedure when contrasted with methods that do not include the trend directly in the model. The effect of the form of the noise (whether constant or heteroscedastic) on the results is also investigated. Methods. We introduced a Bayesian generalised Lomb-Scargle periodogram with trend (BGLST), which is a probabilistic linear regression model using Gaussian priors for the coefficients of the fit and a uniform prior for the frequency parameter. Results. We show, using synthetic data, that when there is no prior information on whether and to what extent the true model of the data contains a linear trend, the introduced BGLST method is preferable to the methods that either detrend the data or opt not to detrend the data before fitting the periodic model. Whether to use noise with other than constant variance in the model depends on the density of the data sampling and on the true noise type of the process.

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