Electron-Microscopic localization of biotinylated gastrin bound to eosinophils in the rat stomach

1994 ◽  
Vol 52 ◽  
pp. 300-301
Author(s):  
Caroline A. Miller ◽  
David H. Nichols ◽  
Richard F. Murphy
Keyword(s):  
Electron Microscope ◽  
Phosphate Buffer ◽  
Uranyl Acetate ◽  
List Type ◽  
Cerium Chloride ◽  
Rat Stomach ◽  
Electron Microscopic ◽  
Human Sequence ◽  
Acetate Lead ◽  
Microscope Slides

Gastrin is a small peptide capable of both stimulating gastric acid secretion and acting as an enteric growth factor. Known functions of eosinophils in the rat stomach are related to immunological defense. Here we demonstrate the binding of biotinylated gastrin to rat stomach eosinophils in the electron microscope. Small pieces of stomach were fixed by immersion in 4% paraformaldehyde/0.1% glutaraldehyde in 0.1 M phosphate buffer, pH 7.4 for 1 hour. The tissue was then cryoprotected in 30% sucrose/0.1 M phosphate buffer, transferred to Tissue Tek OCT compound and frozen in isopentane cooled with liquid nitrogen. Transverse cryostat sections were cut at 25 μm, thawed in PBS and free floating sections exposed to 10−5 M biotinylated 1-17 gastrin (human sequence; Peninsula Labs) for 1 hour. Controls omitted the biotinylated gastrin from this step. Sections were then rinsed 3X in PBS and exposed to either:1).a 1:50 dilution of 10 nm Extravidin colloidal gold (Sigma) for 2 hours, or2).an avidin-biotin-alkaline phosphatase complex (ABC-AP;Vector) for 1 hour. A substrate solution containing cerium chloride was used to generate an electron dense reaction product.Sections from both procedures were postfixed in 1% OsO4 in 0.1 M phosphate buffer, rinsed and dehydrated. These were then flat embedded in EMbed 812 between two microscope slides coated with Liquid Release (both from Electron Microscopy Sciences).Polymerized sections were adhered to resin blocks using super glue, cut at 70-90 nm, stained with uranyl acetate/lead citrate and observed in a Philips CM-10 electron microscope.

Electron Microscopic Studies of Gerbil Testis After Vasectomy

1976 ◽  
Vol 34 ◽  
pp. 154-155
Author(s):  
S. K. MAJUMDAR ◽  
FRED KALENSCHER
Keyword(s):  
Electron Microscope ◽  
Normal Structure ◽  
Uranyl Acetate ◽  
Meriones Unguiculatus ◽  
Mongolian Gerbils ◽  
Electron Microscopic ◽  
Transmission Electron ◽  
Acetate Lead ◽  
Microscopic Studies ◽  
Ultrathin Sections

Ultrathin sections made from bilaterally vasectomized as well as bilaterally sham-operated Mongolian gerbils (Meriones unguiculatus) were examined and compared under an electron microscope in order to determine whether vasectomy has any effect upon the fine structure of the testis. The whole testes were removed and placed in Karnovsky's fixative for one hour. After this period the testes were diced into small pieces and fixed for an additional hour in the same fixative. After rinsing in distilled water and postfixed for one hour in OsO4, the tissues were embedded in Epon 812. The sections were stained with uranyl acetate-lead citrate and examined on a Philips Model 201 transmission electron microscope. Shamoperated testis exhibited normal structure of germ cells.

Buffered Formalin for Primary Fixation and Preserving Tissue for a Long Time

1967 ◽  
Vol 25 ◽  
pp. 24-25
Author(s):  
I. Yamamoto ◽  
B. Rosario
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
Phosphate Buffer ◽  
Standard Technique ◽  
Uranyl Acetate ◽  
Osmic Acid ◽  
List Type ◽  
Thin Sections ◽  
Long Time ◽  
Resin Mixture

It is known that suitably buffered formaldehyde can be used as a killing agent and primary fixative for electron microscopy(Pease).We were interested in how long buffered formalin preserves tissue. Thus, in this study, an attempt was made to observe ultrastructure of surgical material fixed and stored in buffered formalin for a long time.Pieces of fresh tissues were, fixed in buffered formalin at 4°C for one hour, and then placed at room temparature for weeks and months without changing solution.Buffered formalin used was prepared as follows:A. “ reagent grade ” formalinB. 0.05 M Sorensen's phosphate buffer, pH 7.2-7.4 mix 1 part of A and 9 parts of B to make 10 % solution.Pieces of tissues were then minced and postfixed in 1 % osmic acid buffered with veronal acetate for two hours or more. Postfixed tissues were then embedded in an epoxy resin mixture through standard technique. Thin sections were stained with uranyl acetate followed by lead, and examined in a Hitachi 11C electron microscope.

Ultrastructural comparison of prolactin adenomas of pituitary in men and women

1988 ◽  
Vol 46 ◽  
pp. 346-347
Author(s):  
R.C. Caughey ◽  
U.P. Kalyan-Raman
Keyword(s):  
Endoplasmic Reticulum ◽  
Electron Microscope ◽  
Phosphate Buffer ◽  
Pituitary Adenomas ◽  
Osmium Tetroxide ◽  
Secretory Granules ◽  
Uranyl Acetate ◽  
En Bloc ◽  
Men And Women ◽  
Transmission Electron

Prolactin producing pituitary adenomas are ultrastructurally characterized by secretory granules varying in size (150-300nm), abundance of endoplasmic reticulum, and misplaced exocytosis. They are also subclassified as sparsely or densely granulated according to the amount of granules present. The hormone levels in men and women vary, being higher in men; so also the symptoms vary between both sexes. In order to understand this variation, we studied 21 prolactin producing pituitary adenomas by transmission electron microscope. This was out of a total of 80 pituitary adenomas. There were 6 men and 15 women in this group of 21 prolactinomas.All of the pituitary adenomas were fixed in 2.5% glutaraldehyde, rinsed in Millonig's phosphate buffer, and post fixed with 1% osmium tetroxide. They were then en bloc stained with 0.5% uranyl acetate, rinsed with Walpole's non-phosphate buffer, dehydrated with graded series of ethanols and embedded with Epon 812 epoxy resin.

Electron Microscopic Examination of a Transplantable Rat Mammary Carcinoma

1968 ◽  
Vol 26 ◽  
pp. 20-21
Author(s):  
S. Shirahama ◽  
G. C. Engle ◽  
R. M. Dutcher
Keyword(s):  
Electron Microscope ◽  
Osmium Tetroxide ◽  
Electron Microscopic Examination ◽  
Undifferentiated Carcinoma ◽  
Uranyl Acetate ◽  
Sprague Dawley ◽  
Electron Microscopic ◽  
North West ◽  
X Irradiation ◽  
Tissue Specimens

A transplantable carcinoma was established in North West Sprague Dawley (NWSD) rats by use of X-irradiation by Engle and Spencer. The tumor was passaged through 63 generations over a period of 32 months. The original tumor, an adenocarcinoma, changed into an undifferentiated carcinoma following the 19th transplant. The tumor grew well in NWSD rats of either sex at various ages. It was invariably fatal, causing death of the host within 15 to 35 days following transplantation.Tumor, thymus, spleen, and plasma from 7 rats receiving transplants of tumor at 3 to 9 weeks of age were examined with an electron microscope at intervals of 8, 15, 22 and 30 days after transplantation. Four normal control rats of the same age were also examined. The tissues were fixed in glutaraldehyde, postfixed in osmium tetroxide and embedded in Epon. The plasma was separated from heparanized blood and processed as previously described for the tissue specimens. Sections were stained with uranyl acetate followed by lead citrate and examined with an RCA EMU-3G electron microscope.

An Electron Microscopic Investigation of the Pathogenesis of Hemorrhage Induced by Rattlesnake Venom

1974 ◽  
Vol 32 ◽  
pp. 56-57
Author(s):  
Charlotte L. Ownby ◽  
Robert A. Kainer ◽  
Anthony T. Tu
Keyword(s):  
Phosphate Buffer ◽  
Osmium Tetroxide ◽  
Microscopic Investigation ◽  
Uranyl Acetate ◽  
Electron Microscopic Investigation ◽  
Lead Citrate ◽  
Electron Microscopic ◽  
Thin Sections ◽  
Rattlesnake Venom ◽  
Diamondback Rattlesnake

One of the significant changes induced by the injection of rattlesnake (Crotalidae) venom is hemorrhage. Since crotaline antivenin does not prevent such local tissue damage, a more effective treatment of snakebite is needed. To aid in the development of such a treatment the pathogenesis of venom-induced hemorrhae was investigated.Swiss-Webster white mice were injected intramuscularly with Western diamondback rattlesnake (Crotalus atrox) venom. Two minutes after the injection, muscle tissue was obtained by bioosy from the thigh and fixed in 6% glutaraldehyde in Milloniq's phosphate buffer (DH 7.4, 2 hrs., 4°C). After post-fixation in 2% osmium tetroxide in Milloniq's phosphate buffer (pH 7.4, 1hr., 4°C) the tissue was dehydrated routinely in ethanol and embedded in Epon 812. The thin sections were stained with uranyl acetate in methanol and lead citrate then observed with either a Zeiss EM 9A or an Hitachi HS-8 electron microscope.

Electron microscope study on the glial filaments of astrocytes in the rat brain

1978 ◽  
Vol 36 (2) ◽  
pp. 618-619
Author(s):  
S. Mori ◽  
K. Furukawa ◽  
H. Abe
Keyword(s):  
Body Weight ◽  
Electron Microscope ◽  
Glial Cells ◽  
Microscope Study ◽  
Phosphate Buffer ◽  
Electron Microscope Study ◽  
Gold Chloride ◽  
Electron Microscopic ◽  
New Knowledge ◽  
Microscopic Studies

With the electron microscope, it was demonstrated that the glial filaments existed in astrocytes and could be impregnated by Cajal's gold chloride sublimate solution. By this conspicuous structure of glial filaments, thus, the astrocytes have long been differentiated from other glial cells from the classical light microscopic studies till the recent electron microscopic observations. Further investigations could add the new knowledge on this important component of glial cells in this laboratory that the actin- like filaments might be contained among the glial filaments. It will be shown in this report that glial filaments of astrocytes are consisted of the heterogenous groups of filaments, and some of them can bind with the heavy meromyosins (HMM).Normal rats (about 120 g body weight) were anesthetized with Nembutal and fixed by perfusion through the heart for 30 minutes with the fixative. This fluid was consisted of 3 % glutaraldehyde, 2 % paraformaldehyde, 4 % sucrose and 0. 5 mM CaCl2 in 0. 1 M phosphate buffer at PH 7. 4.

Ultrastructural diagnosis of cryptosporidium in a patient with acquired immunodeficiency syndrome (AIDS)

1993 ◽  
Vol 51 ◽  
pp. 418-419
Author(s):  
E. Ramirez ◽  
B. L. A. Ceseñas ◽  
F. Segovia
Keyword(s):  
Electron Microscope ◽  
Acquired Immunodeficiency Syndrome ◽  
Phosphate Buffer ◽  
Uranyl Acetate ◽  
Intestinal Biopsy ◽  
Intestinal Protozoan ◽  
Thin Sections ◽  
Severe Diarrhea ◽  
Acquired Immunodeficiency ◽  
Immunodeficiency Syndrome

Cryptosporidium sp is an intestinal protozoan that is well known as parasite of different animals,but recently it has been recognized as a cause of human disease, princippally since apparition of the acquired immunodeficiency syndrome (AIDS). In this kind of patients it produces severe diarrhea (loss of a stool volume up to 17 liters per day), this condition may be a major factor leading to death.Information about Cryptosporidiosis and AIDS incidence in developing countries is limited. In this paper we report the ultrastructural study of an intestinal biopsy from an hemophilic man,infected wit HIV through a blood transfussion that presented severe diarrhea.Jejunal biopsy specimen was obtained and prepared for electron microscope study. The specimen was fixed in 2.5% glutaraldehyde in 0.2 M phosphate buffer pH 7.2;postfixed in 1% osmium tetroxide in 0.2M phosphate buffer pH 7.2;dehydrated in ethanol and embedded in epoxy resin (Medcast). Thin sections obtained in a LKB ultratome V with a diamond knife, stained with uranyl acetate and lead citrate, were observed in a Zeiss EM109 electron microscope.

Comparison of Cholesterol Extraction from Tissues During Processing for Electron Microscopic Radioautography

1968 ◽  
Vol 26 ◽  
pp. 92-93
Author(s):  
E. Tessa Hedley-Whyte ◽  
Betty G. Uzman
Keyword(s):  
Electron Microscope ◽  
Lipid Metabolism ◽  
Propylene Oxide ◽  
Mouse Liver ◽  
Uranyl Acetate ◽  
Electron Microscopic ◽  
Electron Microscopic Radioautography

Radioautographic studies of lipid metabolism with the electron microscope have been limited by the large losses of lipid which occur during the usual dehydration and infiltration procedures. In this study different methods of processing tissue for electron microscopic radioautography have been compared with respect to the extraction of cholesterol-1, 2-3H from labeled mouse liver and nerve.In all methods primary fixation had been in 10% formalin for several months followed by washing overnight. Post-fixation for one hour in Dalton's chrome osmium and staining in 1% uranyl acetate in 10% formalin resulted in small but constant losses. Four methods were assessed: 1) graded acetone dehydration with propylene oxide and epon-araldite mixture infiltration, 2) exposure to 0.5% digitonin in 50% ethanol for 1 hour before post-fixing in osmium followed by procedure 1, 3) Durcupan embedding, and 4) limited dehydration followed by embedding in an epon:araldite mixture (modified Idelman).

Pressurized Specimen Chamber for Electron Microscope

1970 ◽  
Vol 28 ◽  
pp. 546-547 ◽  
Author(s):  
A. Fukami ◽  
T. Etho ◽  
N. Ishihara ◽  
M. Katoh ◽  
K. Fujiwara
Keyword(s):  
Electron Microscope ◽  
Microscopic Observation ◽  
Electron Microscopic Observation ◽  
Electron Bombardment ◽  
Wide Field ◽  
List Type ◽  
Atmospheric Layer ◽  
Electron Microscopic ◽  
Window Film

Electron microscopic observation is confined to dry specimens because the specimen must be kept in vacuum. To extricate themselves from this restriction, several researchers have developed a pressurized specimen chamber which, however, is not satisfactory in reliability; for example, (1) The Lenard window film is not stable,(2) The atmospheric layer is not so thin as the spacer because of the bend of the Lenard window film,(3) Operation is not easy.Under these circumstances, we have developed a new pressurized specimen chamber, considering the following points; (1) Strength of the Leanard window films against pressure and electron bombardment,(2) Applicable condition for wide field of research, Ease of opration.(3) Ease of opration.

Electron microscopic study on the phagocytic lining cells in the cavernous body of the lamprey gill

1978 ◽  
Vol 36 (2) ◽  
pp. 448-449
Author(s):  
Kazuhito Yamaguchi ◽  
Kazuhiko Awaya
Keyword(s):  
Electron Microscope ◽  
Uranyl Acetate ◽  
Osmic Acid ◽  
Lead Citrate ◽  
Electron Microscopic ◽  
Embedded Blocks ◽  
Ultrathin Sections ◽  
Branchial Artery ◽  
Scanning Electron ◽  
Cavernous Body

The lining cells in the cavernous body, which is a branch of the afferent branchial artery, of lamprey gills were studied with transmission (TEM) and scanning electron microscope (SEM).Adult and larval lampreys, Lampertra planeri, were used in this study. Lamprey gills removed after perfusion were fixed in 2 % glutaraldehyde/formaldehyde (pH 7. 4) for 1 hour and postfixed in 1 % osmic acid. For TEM, the gills were dehydrated in ethanol and embedded in epon 812. Ultrathin sections were doubly stained with uranyl acetate and lead citrate. For SEM, most of the gills were dehydrated in ethanol and isoamylacetate and cracked in liquid nitrogen. Some of the gills dehydrated were embedded in stylene monomer. Stylene embedded blocks were cracked with a hummer and dissolved in propylene oxide. All the specimens were dried in a critical point dryer and coated 100 Å thickness with gold-paladium.The cavernous body is composed of trabeculae, between which are blood spaces, the lacunae.

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