Correlative Fluorescence and Scanning Transmission Electron Microscopy of Quantum Dot-Labeled Proteins on Whole Cells in Liquid

Author(s):  
Diana B. Peckys ◽  
Vera Bandmann ◽  
Niels de Jonge
2016 ◽  
Vol 22 (3) ◽  
pp. 656-665 ◽  
Author(s):  
Justus Hermannsdörfer ◽  
Verena Tinnemann ◽  
Diana B. Peckys ◽  
Niels de Jonge

AbstractWhole cells can be studied in their native liquid environment using electron microscopy, and unique information about the locations and stoichiometry of individual membrane proteins can be obtained from many cells thus taking cell heterogeneity into account. Of key importance for the further development of this microscopy technology is knowledge about the effect of electron beam radiation on the samples under investigation. We used environmental scanning electron microscopy (ESEM) with scanning transmission electron microscopy (STEM) detection to examine the effect of radiation for whole fixed COS7 fibroblasts in liquid. The main observation was the localization of nanoparticle labels attached to epidermal growth factor receptors (EGFRs). It was found that the relative distances between the labels remained mostly unchanged (<1.5%) for electron doses ranging from the undamaged native state at 10 e−/Å2 toward 103 e−/Å2. This dose range was sufficient to determine the EGFR locations with nanometer resolution and to distinguish between monomers and dimers. Various different forms of radiation damage became visible at higher doses, including severe dislocation, and the dissolution of labels.


Author(s):  
F. Khoury ◽  
L. H. Bolz

The lateral growth habits and non-planar conformations of polyethylene crystals grown from dilute solutions (<0.1% wt./vol.) are known to vary depending on the crystallization temperature.1-3 With the notable exception of a study by Keith2, most previous studies have been limited to crystals grown at <95°C. The trend in the change of the lateral growth habit of the crystals with increasing crystallization temperature (other factors remaining equal, i.e. polymer mol. wt. and concentration, solvent) is illustrated in Fig.l. The lateral growth faces in the lozenge shaped type of crystal (Fig.la) which is formed at lower temperatures are {110}. Crystals formed at higher temperatures exhibit 'truncated' profiles (Figs. lb,c) and are bound laterally by (110) and (200} growth faces. In addition, the shape of the latter crystals is all the more truncated (Fig.lc), and hence all the more elongated parallel to the b-axis, the higher the crystallization temperature.


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