scholarly journals Nanoparticle tracking analysis

2020 ◽  
Vol 1 ◽  
Author(s):  
H. Peneder ◽  
E. Punz ◽  
I.A. Joubert ◽  
M. Geppert ◽  
M. Himly

Due to their extremely small size, nanoparticles cannot be analyses by conventional approaches such as light microscopy. To visualise particles in the nanoscale range, a combination of an ultra-microscope and a laser illumination unit has to be applied. This combinatory technique is called Nanoparticle Tracking Anlysis (NTA) and can be used of thr nalysis of particles in a size range of approximately 10 nm up to 1 μm in liquid suspension. 

2017 ◽  
Vol 5 (4) ◽  
Author(s):  
Ciarán M. Maguire ◽  
Katherine Sillence ◽  
Matthias Roesslein ◽  
Claire Hannell ◽  
Guillaume Suarez ◽  
...  

One of the greatest challenges in the manufacturing and development of nanotechnologies is the requirement for robust, reliable, and accurate characterization data. Presented here are the results of an interlaboratory comparison (ILC) brought about through multiple rounds of engagement with NanoSight Malvern and ten pan-European research facilities. Following refinement of the nanoparticle tracking analysis (NTA) technique, the size and concentration characterization of nanoparticles in liquid suspension was proven to be robust and reproducible for multiple sample types in monomodal, binary, or multimodal mixtures. The limits of measurement were shown to exceed the 30–600 nm range (with all system models), with percentage coefficients of variation (% CV) being calculated as sub 5% for monodisperse samples. Particle size distributions were also improved through the incorporation of the finite track length adjustment (FTLA) algorithm, which most noticeably acts to improve the resolution of multimodal sample mixtures. The addition of a software correction to account for variations between instruments also dramatically increased the accuracy and reproducibility of concentration measurements. When combined, the advances brought about during the interlaboratory comparisons allow for the simultaneous determination of accurate and precise nanoparticle sizing and concentration data in one measurement.


2021 ◽  
Vol 22 (8) ◽  
pp. 3839
Author(s):  
Sobha Karuthedom Karuthedom George ◽  
Lucia Lauková ◽  
René Weiss ◽  
Vladislav Semak ◽  
Birgit Fendl ◽  
...  

Growing interest in extracellular vesicles (EVs) has prompted the advancements of protocols for improved EV characterization. As a high-throughput, multi-parameter, and single particle technique, flow cytometry is widely used for EV characterization. The comparison of data on EV concentration, however, is hindered by the lack of standardization between different protocols and instruments. Here, we quantified EV counts of platelet-derived EVs, using two flow cytometers (Gallios and CytoFLEX LX) and nanoparticle tracking analysis (NTA). Phosphatidylserine-exposing EVs were identified by labelling with lactadherin (LA). Calibration with silica-based fluorescent beads showed detection limits of 300 nm and 150 nm for Gallios and CytoFLEX LX, respectively. Accordingly, CytoFLEX LX yielded 40-fold higher EV counts and 13-fold higher counts of LA+CD41+ EVs compared to Gallios. NTA in fluorescence mode (F-NTA) demonstrated that only 9.5% of all vesicles detected in scatter mode exposed phosphatidylserine, resulting in good agreement of LA+ EVs for CytoFLEX LX and F-NTA. Since certain functional characteristics, such as the exposure of pro-coagulant phosphatidylserine, are not equally displayed across the entire EV size range, our study highlights the necessity of indicating the size range of EVs detected with a given approach along with the EV concentration to support the comparability between different studies.


Author(s):  
Nakazo Watari ◽  
Yasuaki Hotta ◽  
Yoshio Mabuchi

It is very useful if we can observe the identical cell elements within the same sections by light microscopy (LM), transmission electron microscopy (TEM) and/or scanning electron microscopy (SEM) sequentially, because, the cell fine structure can not be indicated by LM, while the color is; on the other hand, the cell fine structure can be very easily observed by EM, although its color properties may not. However, there is one problem in that LM requires thick sections of over 1 μm, while EM needs very thin sections of under 100 nm. Recently, we have developed a new method to observe the same cell elements within the same plastic sections using both light and transmission (conventional or high-voltage) electron microscopes.In this paper, we have developed two new observation methods for the identical cell elements within the same sections, both plastic-embedded and paraffin-embedded, using light microscopy, transmission electron microscopy and/or scanning electron microscopy (Fig. 1).


Author(s):  
E.J. Jenkins ◽  
D.S. Tucker ◽  
J.J. Hren

The size range of mineral and ceramic particles of one to a few microns is awkward to prepare for examination by TEM. Electrons can be transmitted through smaller particles directly and larger particles can be thinned by crushing and dispersion onto a substrate or by embedding in a film followed by ion milling. Attempts at dispersion onto a thin film substrate often result in particle aggregation by van der Waals attraction. In the present work we studied 1-10 μm diameter Al2O3 spheres which were transformed from the amprphous state to the stable α phase.After the appropriate heat treatment, the spherical powders were embedded in as high a density as practicable in a hard EPON, and then microtomed into thin sections. There are several advantages to this method. Obviously, this is a rapid and convenient means to study the microstructure of serial slices. EDS, ELS, and diffraction studies are also considerably more informative. Furthermore, confidence in sampling reliability is considerably enhanced. The major negative feature is some distortion of the microstructure inherent to the microtoming operation; however, this appears to have been surprisingly small. The details of the method and some typical results follow.


Author(s):  
C. T. Nightingale ◽  
S. E. Summers ◽  
T. P. Turnbull

The ease of operation of the scanning electron microscope has insured its wide application in medicine and industry. The micrographs are pictorial representations of surface topography obtained directly from the specimen. The need to replicate is eliminated. The great depth of field and the high resolving power provide far more information than light microscopy.


Author(s):  
Bruce Mackay

The broadest application of transmission electron microscopy (EM) in diagnostic medicine is the identification of tumors that cannot be classified by routine light microscopy. EM is useful in the evaluation of approximately 10% of human neoplasms, but the extent of its contribution varies considerably. It may provide a specific diagnosis that can not be reached by other means, but in contrast, the information obtained from ultrastructural study of some 10% of tumors does not significantly add to that available from light microscopy. Most cases fall somewhere between these two extremes: EM may correct a light microscopic diagnosis, or serve to narrow a differential diagnosis by excluding some of the possibilities considered by light microscopy. It is particularly important to correlate the EM findings with data from light microscopy, clinical examination, and other diagnostic procedures.


Author(s):  
Gladys Harrison

With the advent of the space age and the need to determine the requirements for a space cabin atmosphere, oxygen effects came into increased importance, even though these effects have been the subject of continuous research for many years. In fact, Priestly initiated oxygen research when in 1775 he published his results of isolating oxygen and described the effects of breathing it on himself and two mice, the only creatures to have had the “privilege” of breathing this “pure air”.Early studies had demonstrated the central nervous system effects at pressures above one atmosphere. Light microscopy revealed extensive damage to the lungs at one atmosphere. These changes which included perivascular and peribronchial edema, focal hemorrhage, rupture of the alveolar septa, and widespread edema, resulted in death of the animal in less than one week. The severity of the symptoms differed between species and was age dependent, with young animals being more resistant.


Author(s):  
Joseph E. Mazurkiewicz

Immunocytochemistry is a powerful investigative approach in which one of the most exacting examples of specificity, that of the reaction of an antibody with its antigen, isused to localize tissue and cell specific molecules in situ. Following the introduction of fluorescent labeled antibodies in T950, a large number of molecules of biological interest had been studied with light microscopy, especially antigens involved in the pathogenesis of some diseases. However, with advances in electron microscopy, newer methods were needed which could reveal these reactions at the ultrastructural level. An electron dense label that could be coupled to an antibody without the loss of immunologic activity was desired.


Author(s):  
E. S. Boatman ◽  
G. E. Kenny

Information concerning the morphology and replication of organism of the family Mycoplasmataceae remains, despite over 70 years of study, highly controversial. Due to their small size observations by light microscopy have not been rewarding. Furthermore, not only are these organisms extremely pleomorphic but their morphology also changes according to growth phase. This study deals with the morphological aspects of M. pneumoniae strain 3546 in relation to growth, interaction with HeLa cells and possible mechanisms of replication.The organisms were grown aerobically at 37°C in a soy peptone yeast dialysate medium supplemented with 12% gamma-globulin free horse serum. The medium was buffered at pH 7.3 with TES [N-tris (hyroxymethyl) methyl-2-aminoethane sulfonic acid] at 10mM concentration. The inoculum, an actively growing culture, was filtered through a 0.5 μm polycarbonate “nuclepore” filter to prevent transfer of all but the smallest aggregates. Growth was assessed at specific periods by colony counts and 800 ml samples of organisms were fixed in situ with 2.5% glutaraldehyde for 3 hrs. at 4°C. Washed cells for sectioning were post-fixed in 0.8% OSO4 in veronal-acetate buffer pH 6.1 for 1 hr. at 21°C. HeLa cells were infected with a filtered inoculum of M. pneumoniae and incubated for 9 days in Leighton tubes with coverslips. The cells were then removed and processed for electron microscopy.


Author(s):  
N.C. Lyon ◽  
W. C. Mueller

Schumacher and Halbsguth first demonstrated ectodesmata as pores or channels in the epidermal cell walls in haustoria of Cuscuta odorata L. by light microscopy in tissues fixed in a sublimate fixative (30% ethyl alcohol, 30 ml:glacial acetic acid, 10 ml: 65% nitric acid, 1 ml: 40% formaldehyde, 5 ml: oxalic acid, 2 g: mecuric chloride to saturation 2-3 g). Other workers have published electron micrographs of structures transversing the outer epidermal cell in thin sections of plant leaves that have been interpreted as ectodesmata. Such structures are evident following treatment with Hg++ or Ag+ salts and are only rarely observed by electron microscopy. If ectodesmata exist without such treatment, and are not artefacts, they would afford natural pathways of entry for applied foliar solutions and plant viruses.


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