Simulation-based optimisation of thermodynamic conditions in the ESEM for dynamical in-situ study of spherical polyelectrolyte complex particles in their native state

Neděla, Vilém; Tihlaříková, Eva; Maxa, Jiří; Imrichová, Kamila; Bučko, Marek; Gemeiner, Peter

doi:10.1016/j.ultramic.2020.112954

Cited by 29 publications

(24 citation statements)

References 57 publications

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“…Further optimization of the conditions will be carried out after analyzing results with the software TDS (Num Solution & ISI CAS, Czech Republic), which is based on ANSYS software simulations and online in situ measurement of the sample temperature and environmental humidity. Measurements will be performed via micro-sensors integrated into the self-developed Peltier cooling stage in the A-ESEM [49]. This all depends on the optimization of staining procedures for plant cells that would utilize in vivo labeling with antibodies or specific labeling with fluorescent ligands carrying nanogold or quantum dots [50].…”

Section: Discussionmentioning

confidence: 99%

Correlative Light-Environmental Scanning Electron Microscopy of Plasma Membrane Efflux Carriers of Plant Hormone Auxin

et al. 2021

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Fluorescence light microscopy provided convincing evidence for the domain organization of plant plasma membrane (PM) proteins. Both peripheral and integral PM proteins show an inhomogeneous distribution within the PM. However, the size of PM nanodomains and protein clusters is too small to accurately determine their dimensions and nano-organization using routine confocal fluorescence microscopy and super-resolution methods. To overcome this limitation, we have developed a novel correlative light electron microscopy method (CLEM) using total internal reflection fluorescence microscopy (TIRFM) and advanced environmental scanning electron microscopy (A-ESEM). Using this technique, we determined the number of auxin efflux carriers from the PINFORMED (PIN) family (NtPIN3b-GFP) within PM nanodomains of tobacco cell PM ghosts. Protoplasts were attached to coverslips and immunostained with anti-GFP primary antibody and secondary antibody conjugated to fluorochrome and gold nanoparticles. After imaging the nanodomains within the PM with TIRFM, the samples were imaged with A-ESEM without further processing, and quantification of the average number of molecules within the nanodomain was performed. Without requiring any post-fixation and coating procedures, this method allows to study details of the organization of auxin carriers and other plant PM proteins.

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Section: Discussionmentioning

confidence: 99%

Correlative Light-Environmental Scanning Electron Microscopy of Plasma Membrane Efflux Carriers of Plant Hormone Auxin

et al. 2021

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“…The most accurate shape which copies the gas expansion is the Laval nozzle. This can be constructed in three ways: characteristic method, linear shape, or Bell nozzle ( Figure 6 , Figure 7 and Figure 8 ) [ 7 ]. Due to the production technology, the most advantageous construction is based on linear shape ( Figure 8 ).…”

Section: Static Pressure Measurement According To Prandtl’s Theorymentioning

confidence: 99%

“…The pressure and shock wave distribution in the primary path of the electron beam passing through the differentially pumped chamber has a great influence on the resulting image quality [ 1 , 2 , 3 ]. The environmental scanning electron microscope (ESEM) is one of the most promising tools for studying plant [ 4 , 5 , 6 ] and polymer [ 7 , 8 ] samples using special signal electron detectors [ 9 ], which were developed based on gas flow simulations.…”

Section: Introductionmentioning

confidence: 99%

Application of Prandtl’s Theory in the Design of an Experimental Chamber for Static Pressure Measurements

Šabacká

Neděla

Maxa

et al. 2021

Sensors

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Pumping in vacuum chambers is part of the field of environmental electron microscopy. These chambers are separated from each other by a small-diameter aperture that creates a critical flow in the supersonic flow regime. The distribution of pressure and shock waves in the path of the primary electron beam passing through the differentially pumped chamber has a large influence on the quality of the resulting microscope image. As part of this research, an experimental chamber was constructed to map supersonic flow at low pressures. The shape of this chamber was designed using mathematical–physical analyses, which served not only as a basis for the design of its geometry, but especially for the correct choice of absolute and differential pressure sensors with respect to the cryogenic temperature generated in the supersonic flow. The mathematical and physical analyses presented here map the nature of the supersonic flow with large gradients of state variables at low pressures at the continuum mechanics boundary near the region of free molecule motion in which the Environmental Electron Microscope and its differentially pumped chamber operate, which has a significant impact on the resulting sharpness of the final image obtained by the microscope. The results of this work map the flow in and behind the Laval nozzle in the experimental chamber and are the initial basis that enabled the optimization of the design of the chamber based on Prandtl’s theory for the possibility of fitting it with pressure probes in such a way that they can map the flow in and behind the Laval nozzle.

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“…The ESEM is suitable for the direct imaging of wet, electrically non-conductive and electron beam-sensitive samples (Michaloudi et al, 2018) at chamber pressures up to 2,000 Pa; the samples do not require conductive coating (Schenkmayerová et al, 2014). Moreover, the device allows in-situ observation under dynamically changing environmental conditions (Neděla et al, 2020). The samples are placed on the silicon pad of a Peltier cooling stage, which can reach temperatures down to −27 °C; this ESEM facilitates the imaging of frozen samples.…”

Section: Description Of the Esemmentioning

confidence: 99%

Sublimation of frozen CsCl solutions in ESEM: determining the number and size of salt particles relevant to sea-salt aerosols

Vetráková

Neděla²,

Runštuk³

et al. 2022

Preprint

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Abstract. We identified the factors that, during the sublimation of a frozen CsCl solution, are important in generating fine salt particles as a possible source of salt aerosol. The number, size, and structure of the particles that remain after ice sublimation were investigated with respect to the concentration of the salt in the sample, the freezing method, and the sublimation temperature. The last-named aspect is evidently of primary importance for the preference of fine salt crystals over a large compact piece of salt. Independently of the concentration and freezing method tested, the sublimation of the frozen samples above the eutectic temperature (Teu) yielded a large compact piece of salt, namely, an improbable source of aerosol particles. Small salt particles that might be a source of atmospheric aerosols were formed predominantly at the temperatures below Teu, and their structures strongly depended on the concentration of the salt. For example, the sublimation of those samples that exhibited less than 8 psu (0.05 M) often produced small aerosolizable isolated particles readily able to be windblown. Conversely, the sublimation of 78 psu (0.5 M) samples led to the formation of relatively stable and largely interconnected salt structures. Presumably, our findings have important implications because the formed salt particles may assume the role of cloud condensation nuclei and ice-nucleating particles, affect polar atmospheric radiative forcing, and facilitate heterogeneous atmospheric chemistry.

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Simulation-based optimisation of thermodynamic conditions in the ESEM for dynamical in-situ study of spherical polyelectrolyte complex particles in their native state

Cited by 29 publications

References 57 publications

Correlative Light-Environmental Scanning Electron Microscopy of Plasma Membrane Efflux Carriers of Plant Hormone Auxin

Correlative Light-Environmental Scanning Electron Microscopy of Plasma Membrane Efflux Carriers of Plant Hormone Auxin

Application of Prandtl’s Theory in the Design of an Experimental Chamber for Static Pressure Measurements

Sublimation of frozen CsCl solutions in ESEM: determining the number and size of salt particles relevant to sea-salt aerosols

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