New versatile staining reagents for biological transmission electron microscopy that substitute for uranyl acetate

2011 ◽  
Vol 60 (6) ◽  
pp. 401-407 ◽  
Author(s):  
M. Nakakoshi ◽  
H. Nishioka ◽  
E. Katayama
Author(s):  
John H. L. Watson ◽  
Jessica Goodwin ◽  
E. Osborne Coates

Biopsies of lung were taken at operation from a patient with semi-acute diffuse pulmonary infiltrates for study by TEM and SEM. Tissue by light microscopy showed non-caseating granulomas consistent with sarcoidosis. Clinical evidence suggested a hypersensitivity reaction related to inhalation of substance of undetermined nature. Samples were fixed in glutaraldehyde, cacodylate-buffered. They were critical point dried and coated with Au-Pd for SEM, and were handled appropriately for TEM in Araldite. Sections were contrasted with uranyl acetate and lead citrate.


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.


Author(s):  
R.H.M. Cross ◽  
A.N. Hodgson ◽  
R.T.F. Bernard

Uranyl acetate is routinely used in the staining of thin sections of biological tissue for transmission electron microscopy. Although many methods for its preparation and use have been described, there is seldom reference to the reasons for variations in concentration, solvent, storage time and staining time. Likewise, possible variations in the effects of staining under different conditions are largely ignored. In order to gain clarity on this issue an attempt has been made to test three variables (solvent, storage time and use in light or dark) under controlled experimental conditions.The tissues used for the experiment were the testis of a marine limpet, the gut epithelium of a fresh-water catfish, and the kidney of a rat; all of which were fixed and embedded by standard methods. The uranyl acetate solutions were prepared at the outset of the experiment and dispensed into small volumes and stored in the dark at 4°C until required.


2007 ◽  
Vol 70 (1) ◽  
pp. 43-49 ◽  
Author(s):  
Sumire Inaga ◽  
Tetsuo Katsumoto ◽  
Keiichi Tanaka ◽  
Toshio Kameie ◽  
Hironobu Nakane ◽  
...  

1998 ◽  
Vol 4 (S2) ◽  
pp. 1138-1139
Author(s):  
G. M. Vernon ◽  
E. J. Rappa ◽  
W. C. Murray ◽  
R. Witkus

Crustacean hemocytes have been characterized on the basis of cell size and nature of cytoplasmic granules. Based on light microscopic morphological analysis and cytochemistry, investigators variously named the hemocyte types (agranular, small-granule, large granule, undifferentiated, hyaline cells, non-explosive, explosive granulocytes, etc.). In his study of the isopod Armadillidium vulgare Faso adopted the terminology of Benjamin and James and referred to the hemocytes as hyaline cells, semi-granulocytes and granulocytes.In the present investigation we have studied the hemocytes of two isopods, Oniscus asellus and Armadillidium nasatum, using transmission electron microscopy. Hemolymph was collected by penetrating the posterior dorsal exoskeleton of 20 animals of each genus with a microcapillary pipette and drawing 3-5μL per isopod. The samples were processed following a standard technique. Thin sections were collected on 300 mesh copper grids, counterstained with 2% aqueous uranyl acetate and lead citrate, and viewed with a JEOL 1010 electron microscope.


1999 ◽  
Vol 276 (4) ◽  
pp. L631-L641 ◽  
Author(s):  
Nades Palaniyar ◽  
Ross A. Ridsdale ◽  
Stephen A. Hearn ◽  
Yew Meng Heng ◽  
F. Peter Ottensmeyer ◽  
...  

Pulmonary surfactant, a mixture of lipids and surfactant proteins (SPs), plays an important role in respiration and gas exchange. SP-A, the major SP, exists as an octadecamer that can self-associate to form elongated protein filaments in vitro. We have studied here the association of purified bovine SP-A with lipid vesicle bilayers in vitro with negative staining with uranyl acetate and transmission electron microscopy. Native bovine surfactant was also examined by transmission electron microscopy of thinly sectioned embedded material. Lipid vesicles made from dipalmitoylphosphatidylcholine and egg phosphatidylcholine (1:1 wt/wt) generally showed a smooth surface morphology, but some large vesicles showed a corrugated one. On the smooth-surfaced vesicles, SP-As primarily interacted in the form of separate octadecamers or as multidirectional protein networks. On the surfaces of the striated vesicles, SP-As primarily formed regularly spaced unidirectional filaments. The mean spacing between adjacent striations and between adjacent filaments was 49 nm. The striated surfaces were not essential for the formation of filaments but appeared to stabilize them. In native surfactant preparations, SP-A was detected in the dense layers. This latter arrangement of the lipid bilayer-associated SP-As supported the potential relevance of the in vitro structures to the in vivo situation.


Parasitology ◽  
1984 ◽  
Vol 89 (1) ◽  
pp. 71-78 ◽  
Author(s):  
B. Leitch ◽  
A. J. Probert ◽  
N. W. Runham

SummaryThe ultrastructure of the tegument of Schistosoma haematobium was examined using scanning and transmission electron microscopy. The surface of the male worm is characterized by numerous raised tubercles bearing apically directed spines. The female in contrast to the male is cylindrical and relatively smooth. Details of oral and ventral suckers are given. The use of uranyl acetate as a tertiary fixative and en bloc stain has revealed the heptalaminate nature of the outer membrane. Tegumental mitochondria are shown to be morphologically more complex than those of S. mansoni. Spherical and elliptical inclusion bodies are also described. The ultrastructure of the oesophageal tegument of S. haematobium is described for the first time and corresponds with earlier observations of S. mansoni.


Author(s):  
M.A. Gillott ◽  
G. Erdös ◽  
D. E. Buetow

Mesophyll cells isolated from soybeans (Glycine max, L. Merr. var. Corsoy) can be grown photoautotrophically in suspension culture. The SB-P cell line can be bleached by maintaining them in total darkness in sucrose supplemented media for several weeks, and will regain photosynthetic competency when returned to the light. This system is ideally suited for the study of gene regulation and the biochemical and ultrastructural changes which occur during the greening process.Cells were fixed for electron microscopy after 8 weeks of growth in total darkness and at intervals of 1 h to 12 d after transfer to the light. Chlorophyll measurements were determined for each sample. For transmission electron microscopy, the cells were fixed for 1-2 h in 4% glutaraldehyde in 0.1M Pipes buffer, pH 7.4, washed in the same buffer, then postfixed for 1 h in 1% OsO4 in Pipes, pH 6.8. Following a graded ethanol dehydration series the cells were transferred into propylene oxide and embedded in Epon. Sections were stained with uranyl acetate and observed on a JEOL 100C TEM operated at 80 kV.


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