An Overview of Novel and Conventional Methods to Detect Extracellular Vesicles

2014 ◽  
pp. 125-156 ◽  
2021 ◽  
Vol 14 (4) ◽  
pp. 356
Author(s):  
Sara Busatto ◽  
Dalila Iannotta ◽  
Sierra A. Walker ◽  
Luisa Di Marzio ◽  
Joy Wolfram

Extracellular vesicles (EVs) mediate intercellular transport of biomolecular cargo in the body, making them promising delivery vehicles for bioactive compounds. Genetic engineering of producer cells has enabled encapsulation of therapeutic proteins in EVs. However, genetic engineering approaches can be expensive, time-consuming, and incompatible with certain EV sources, such as human plasma and bovine milk. The goal of this study was to develop a quick, versatile, and simple method for loading proteins in EVs post-isolation. Proteins, including CRISPR associated protein 9 (Cas9), were bound to cationic lipids that were further complexed with MDA-MB-231 cell-derived EVs through passive incubation. Size-exclusion chromatography was used to remove components that were not complexed with EVs. The ability of EVs to mediate intracellular delivery of proteins was compared to conventional methods, such as electroporation and commercial protein transfection reagents. The results indicate that EVs retain native features following protein-loading and obtain similar levels of intracellular protein delivery as conventional methods, but display less toxicity. This method opens up opportunities for rapid exploration of EVs for protein delivery.


Author(s):  
Kazuyuki Koike ◽  
Hideo Matsuyama

Spin-polarized scanning electron microscopy (spin SEM), where the secondary electron spin polarization is used as the image signal, is a novel technique for magnetic domain observation. Since its first development by Koike and Hayakawa in 1984, several laboratories have extensively studied this technique and have greatly improved its capability for data extraction and its range of applications. This paper reviews the progress over the last few years.Almost all the high expectations initially held for spin SEM have been realized. A spatial resolution of several hundreds angstroms has been attained, which is nearly one order of magnitude higher than that of conventional methods for thick samples. Quantitative analysis of magnetization direction has been performed more easily than with conventional methods. Domain observation of the surface of three-dimensional samples has been confirmed to be possible. One of the drawbacks, a long image acquisition time, has been eased by combining highspeed image-signal processing with high speed scanning, although at the cost of image quality. By using spin SEM, the magnetic structure of a 180 degrees surface Neel wall, magnetic thin films, multilayered films, magnetic discs, etc., have been investigated.


2012 ◽  
Vol 28 (1) ◽  
pp. 11-18 ◽  
Author(s):  
Marcus Roth ◽  
Philipp Hammelstein

Based on the conception of sensation seeking as a need rather than a temperamental trait ( Hammelstein, 2004 ), we present a new assessment method, the Need Inventory of Sensation Seeking (NISS), which is considered to assess a motivational disposition. Three studies are presented: The first examined the factorial structure and the reliability of the German versions of the NISS; the second study compared the German and the English versions of the NISS; and finally, the validity of the NISS was examined in a nonclinical study and compared to the validity of conventional methods of assessing sensation seeking (Sensation Seeking Scale – Form V; SSS-V). Compared to the SSS-V, the NISS shows better reliability and validity in addition to providing new research possibilities including application in experimental areas.


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