Adsorption Staining Methd For Ultrathin Frozen Sections

Author(s):  
K. T. Tokuyasu

The successful routine use of cryoultramicrotomy to examine ultrathin sections by transmission electron microscopy requires the application of suitable staining to delineate the ultrastructure. While negative staining is quite effective for certain purposes1, 2, positive staining is more appropriate for immunocytochemical studies because it does not obscure the immunolabels.

Author(s):  
Kenichi Takaya

Mast cell and basophil granules of the vertebrate contain heparin or related sulfated proteoglycans. Histamine is also present in mammalian mast cells and basophils. However, no histamine is detected in mast cell granules of the amphibian or fish, while it is shown in those of reptiles and birds A quantitative x-ray microanalysis of mast cell granules of fresh frozen dried ultrathin sections of the tongue of Wistar rats and tree frogs disclosed high concentrations of sulfur in rat mast cell granules and those of sulfur and magnesium in the tree frog granules. Their concentrations in tree frog mast cell granules were closely correlated (r=0.94).Fresh frozen dried ultrathin sections and fresh air-dried prints of the tree frog tongue and spleen and young red-eared turtle (ca. 6 g) spleen and heart blood were examined by a quantitative energy-dispersive x-ray microanalysis (X-650, Kevex-7000) for the element constituents of the granules of mast cells and basophils. The specimens were observed by transmission electron microscopy (TEM) (80-200 kV) and followed by scanning transmission electron microscopy (STEM) under an analytical electron microscope (X-650) at an acceleration voltage of 40 kV and a specimen current of 0.2 nA. A spot analysis was performed in a STEM mode for 100 s at a specimen current of 2 nA on the mast cell and basophil granules and other areas of the cells. Histamine was examined by the o-phthalaldehyde method.


2010 ◽  
Vol 16 (4) ◽  
pp. 456-461 ◽  
Author(s):  
Valentina Bello ◽  
Giovanni Mattei ◽  
Paolo Mazzoldi ◽  
Nicoletta Vivenza ◽  
Paolo Gasco ◽  
...  

AbstractLipid-containing nanostructures, in the form of solid lipid nanoparticles or iron oxide nanoparticles (NPs) coated with a lipid shell, were used as case studies for assessing and optimizing staining for transmission electron microscopy structural and compositional characterization. These systems are of paramount importance as drug delivery systems or as bio-compatible contrast agents. In particular, we have treated the systems with a negative (phospshotungstic acid) or with a positive (osmium tetroxide) staining agent. For iron-oxide NPs coated with the lipid shell, negative staining was more efficient with respect to the positive one. Nevertheless, in particular cases the combination of the two staining procedures provided more complete morphological and compositional characterization of the particles.


1996 ◽  
Vol 44 (1) ◽  
pp. 69
Author(s):  
LRG Cannon ◽  
NA Warson

Temnocephala minor Haswell, 1888 lives ectosymbiotically on the surface of the freshwater crayfish Cherax destructor in the Murray-Darling drainages of Australia. Some glands open on the postero-lateral margin and, being moderately refractory to many stains, can be overlooked in whole mounts and sections, and were, in fact, missed by Haswell. Observations were made on living worms with intra vitam dyes, and on whole mounts, wax sections and ultrathin sections using transmission electron microscopy (TEM) to characterise the secretion from these glands and ascertain its mode of manufacture. The function of the glands remains unknown although it appears non-adhesive.


2014 ◽  
Vol 875-877 ◽  
pp. 695-698
Author(s):  
Qian Li ◽  
Li Juan Zhang ◽  
Fang Yang ◽  
Wen Li Zhang ◽  
Hong Xu

ts hard to get ideal ultrathin sections because of the adamant SiO2 dust in silicosis, after perfusion fixation methods and strict control of the cutting speed, improving the success rate of the Silicosis tissue TEM sample preparation of ultrathin sections,so we can more clearly and accurately observed ultrastructural changes of silicosis,and it also can offer morphological basis for research the silicosis organizations function histological changes.


2018 ◽  
Vol 24 (S1) ◽  
pp. 1730-1731 ◽  
Author(s):  
Kelly A. Parker ◽  
Yue Li ◽  
Adam Eshein ◽  
Shengshuang Zhu ◽  
Cheri M. Hampton ◽  
...  

Author(s):  
O. L. Shaffer ◽  
M. S. El-Aasser ◽  
J. W. Vanderhoff

Measurement of particle size and particle morphology by transmission electron microscopy (TEM) is important in the characterization of polymer latex systems. Special sample treatment methods have been developed to permit the study of latexes that present problems, such as low-glass-transition-temperature (Tg) and electron-transparent particles. Some of these methods include hardening and staining by osmium tetroxide or bromine of latexes that contain unsaturation, negative staining by uranyl acetate, and freezing of latexes with low glass-transition temperatures.We have recently found phosphotungstic acid (PTA) to be useful in both negative and positive staining of latex particles. As a negative stain, PTA can enhance the contrast between the electron-transparent particles and the dense PTA-stained background. This has been particularly useful with latexes such as poly(butyl acrylate), poly(ethyl acrylate), poly(ethylene), and other polymers of similar electron densities. As a positive stain, PTA can also react with the surface functional groups of particles, such as hydroxyl, carboxyl, and amine groups.


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