scholarly journals A method for identifying peripheral connections of perivascular nerves based on sensitive acetylcholinesterase staining via perfusion.

1994 ◽  
Vol 42 (2) ◽  
pp. 223-230 ◽  
Author(s):  
R L Bleys ◽  
G J Groen ◽  
M A Matthijssen

To identify peripheral connections of perivascular nerves, which are usually hard to find and easily damaged, we developed a sensitive method for acetylcholinesterase (AChE) staining in rat. The procedure is based on primary staining via perfusion, which permits visualization of perivascular nerves before dissection and, after dissection, further staining via immersion for identification of peripheral connections of the earlier stained perivascular nerves. The AChE histochemistry is based on an intensification method originally described for sections. The procedure has been optimized for staining via vascular perfusion and can also be applied on whole-mount preparations via immersion. The results show intense nerve staining with minimal background. Although the procedure was initially developed for cerebrovascular nerves, the staining approach can also be applied in other regions, the main advantage being that perivascular neural structures can be identified without the damage that usually occurs during dissection.

1988 ◽  
Vol 182 (2) ◽  
pp. 155-168 ◽  
Author(s):  
Shigeru Kuratani ◽  
Shigenori Tanaka ◽  
Yuji Ishikawa ◽  
Chosei Zukeran

2010 ◽  
Vol 32 (1) ◽  
pp. 37-46
Author(s):  
원성윤 ◽  
Mi-Sun Hur ◽  
김희진 ◽  
Jongtae Park ◽  
곽현호 ◽  
...  

Author(s):  
Peter M. Andrews

Although there have been a number of recent scanning electron microscopic reports on the renal glomerulus, the advantages of scanning electron microscopy have not yet been applied to a systematic study of the uriniferous tubules. In the present investigation, scanning electron microscopy was used to study the ultrastructural morphology of the proximal, distal, thin loop, and collecting tubules. Material for observation was taken from rat kidneys which were fixed by vascular perfusion, sectioned by either cutting or fracturing technigues, and critically point dried.The brush border characterising proximal tubules is first detected on the luminal surface of Bowman's capsule adjacent to the urinary pole orifice. In this region one frequently finds irregular microvilli characterized by broad and flattened bases with occasional bulbous structures protruding from their surfaces.


Author(s):  
E. T. O'Toole ◽  
R. R. Hantgan ◽  
J. C. Lewis

Thrombocytes (TC), the avian equivalent of blood platelets, support hemostasis by aggregating at sites of injury. Studies in our lab suggested that fibrinogen (fib) is a requisite cofactor for TC aggregation but operates by an undefined mechanism. To study the interaction of fib with TC and to identify fib receptors on cells, fib was purified from pigeon plasma, conjugated to colloidal gold and used both to facilitate aggregation and as a receptor probe. Described is the application of computer assisted reconstruction and stereo whole mount microscopy to visualize the 3-D organization of fib receptors at sites of cell contact in TC aggregates and on adherent cells.Pigeon TC were obtained from citrated whole blood by differential centrifugation, washed with Ca++ free Hank's balanced salts containing 0.3% EDTA (pH 6.5) and resuspended in Ca++ free Hank's. Pigeon fib was isolated by precipitation with PEG-1000 and the purity assessed by SDS-PAGE. Fib was conjugated to 25nm colloidal gold by vortexing and the conjugates used as the ligand to identify fib receptors.


Author(s):  
R. P. Becker ◽  
J. J. Wolosewick ◽  
J. Ross-Stanton

Methodology has been introduced recently which allows transmission and scanning electron microscopy of cell fine structure in semi-thin sections unencumbered by an embedding medium. Images obtained from these “resinless” sections show a three-dimensional lattice of microtrabeculfee contiguous with cytoskeletal structures and membrane-bounded cell organelles. Visualization of these structures, especially of the matiiDra-nous components, can be facilitated by employing tannic acid in the fixation step and dessicator drying, as reported here.Albino rats were fixed by vascular perfusion with 2% glutaraldehyde or 1.5% depolymerized paraformaldehyde plus 2.5% glutaraldehyde in 0.1M sodium cacodylate (pH 7.4). Tissues were removed and minced in the fixative and stored overnight in fixative containing 4% tannic acid. The tissues were rinsed in buffer (0.2M cacodylate), exposed to 1% buffered osmium tetroxide, dehydrated in ethyl alcohol, and embedded in pure polyethylene glycol-6000 (PEG). Sections were cut on glass knives with a Sorvall MT-1 microtome and mounted onto poly-L-lysine, formvar-carbon coated grids while submerged in a solution of 95% ethanol containing 5% PEG.


Author(s):  
T. L. Hayes

Biomedical applications of the scanning electron microscope (SEM) have increased in number quite rapidly over the last several years. Studies have been made of cells, whole mount tissue, sectioned tissue, particles, human chromosomes, microorganisms, dental enamel and skeletal material. Many of the advantages of using this instrument for such investigations come from its ability to produce images that are high in information content. Information about the chemical make-up of the specimen, its electrical properties and its three dimensional architecture all may be represented in such images. Since the biological system is distinctive in its chemistry and often spatially scaled to the resolving power of the SEM, these images are particularly useful in biomedical research.In any form of microscopy there are two parameters that together determine the usefulness of the image. One parameter is the size of the volume being studied or resolving power of the instrument and the other is the amount of information about this volume that is displayed in the image. Both parameters are important in describing the performance of a microscope. The light microscope image, for example, is rich in information content (chemical, spatial, living specimen, etc.) but is very limited in resolving power.


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