scholarly journals Application of Cationic Colloidal Gold Staining for the Demonstration of Anionic Sites.

1998 ◽  
Vol 31 (1) ◽  
pp. 1-8 ◽  
Author(s):  
Kenji Saga
Keyword(s):  
Colloidal Gold ◽  
Anionic Sites ◽  
Cationic Colloidal Gold

Visualization of the anionic sites in the cell wall of apple fruit using a cationic colloidal gold probe

1993 ◽  
Vol 51 ◽  
pp. 320-321
Author(s):  
Stéphane Roy ◽  
William S. Conway ◽  
Alley E. Watada ◽  
Christopher D. Pooley ◽  
William P. Wergin
Keyword(s):  
Cell Wall ◽  
Colloidal Gold ◽  
Apple Fruit ◽  
Gold Complex ◽  
Anionic Sites ◽  
Wall Strength ◽  
Anionic Binding Sites ◽  
Cationic Colloidal Gold ◽  
Softening Process ◽  
Gold Probe

The ripening of fleshy fruits involves a softening process that consists of biochemical changes in the cell wall and leads to cell separation. Calcium is an important constituent of the cell wall and plays roles in maintaining the firmness of fruit and in reducing postharvest decay. The modification of cell wall strength is believed to be influenced by calcium that interacts with acidic pectic polymers to form crossbridges. This study examined how the frequency and distribution of anionic binding sites in the cell walls of apple fruit were influenced by calcium infiltration.Mature “Golden Delicious” apple fruits were pressure infiltrated with either H2O or a 4% solution of CaCl2 and the pericarp was sampled and processed according to standard procedures. Cationic poly-Llysine colloidal gold complex was used in a one-step procedure to visualize anionic sites in muro. Observations were performed with light microscopy, following silver intensification, and with transmission electron microscopy.

scholarly journals Demonstration of anionic sites in human eccrine and apocrine sweat glands in post-embedded ultra-thin sections with cationic colloidal gold: effect of enzyme digestion on these anionic sites.

1993 ◽  
Vol 41 (8) ◽  
pp. 1197-1207 ◽  
Author(s):  
K Saga ◽  
M Takahashi
Keyword(s):  
Colloidal Gold ◽  
Dermatan Sulfate ◽  
Sweat Glands ◽  
Secretory Cells ◽  
Anionic Sites ◽  
Chondroitinase Abc ◽  
Dark Cell ◽  
Luminal Membrane ◽  
Cationic Colloidal Gold ◽  
Eccrine Sweat Glands

We localized anionic sites ultrastructurally in human eccrine and apocrine sweat glands with a poly-L-lysine-gold complex (cationic colloidal gold). Anionic sites were labeled by incubating Lowicryl K4M-embedded sections on droplets of cationic colloidal gold. In eccrine sweat glands, colloidal gold particles were restricted to the basolateral membrane of the secretory cells at low pH, whereas the luminal membrane did not react with the gold particles. Chondroitinase ABC digested these anionic sites. This indicates that chondroitin sulfate and/or dermatan sulfate constitutes anionic sites in the basal labyrinth of eccrine sweat glands. In apocrine sweat glands, the luminal membrane of the secretory cells showed strong reaction at low pH, whereas the contraluminal membrane did not show any reaction. Neuraminidase completely digested these anionic sites, which indicated that the anionic charge of the apocrine lumen was due to sialic acid. Differences in distribution and susceptibility to enzymes of anionic sites in cell membranes between eccrine and apocrine sweat glands may reflect functional differences between these glands. Dark cell granules in eccrine secretory cells were negative for the anionic sites when sections were labeled without any pre-treatment. However, pre-incubation of the grids on EGTA or deionized water unmasked the anionic sites on the dark cell granules. The positive staining after EGTA treatment was greatly decreased by reincubation with CaCl2. These results suggested that Ca blocked anionic sites in dark cell granules. Exposed anionic sites were digested with chondroitinase ABC. This indicated that chondroitinase ABC and/or dermatan sulfate composed the anionic sites in dark cell granules.

scholarly journals Detection of glomerular anionic sites in post-embedded ultra-thin sections using cationic colloidal gold.

1991 ◽  
Vol 39 (7) ◽  
pp. 965-972 ◽  
Author(s):  
N P Goode ◽  
M Shires ◽  
D M Crellin ◽  
A M Davison
Keyword(s):  
Hyaluronic Acid ◽  
Chondroitin Sulfate ◽  
Colloidal Gold ◽  
Endothelial Glycocalyx ◽  
Computer Assisted ◽  
Nuclear Staining ◽  
Anionic Sites ◽  
Thin Sections ◽  
Cationic Colloidal Gold ◽  
Influence Of Ph

We detected glomerular anionic sites in fixed, LR Gold-embedded ultra-thin tissue sections using cationic colloidal gold. Manual and computer-assisted quantitation were compared, and the influence of pH and glycosaminoglycan-degrading enzymes on site expression was examined. Both quantitation methods produced similar results. Alteration of pH within a narrow range (pH 2.5-3.0) markedly affected the staining pattern. At pH 2.5, epithelial and endothelial glycocalyx and regular sites restricted to the lamina rara externa were stained. At pH 3.0 and above, glycocalyx was unstained but intracellular and nuclear staining was present; glomerular basement membrane (GBM) and mesangial matrix sites were abundant. After chondroitinase ABC or hyaluronidase digestion, GBM staining was eliminated at pH 2.0 and reduced at pH 7.0 (p less than 0.001), suggesting that degraded sites are associated with chondroitin sulfate or hyaluronic acid. By contrast, prolonged heparitinase I digestion was ineffective at either pH. Digestion of purified substrates revealed crossreactivity of heparitinase towards chondroitin sulfate and of chondroitinase towards hyaluronic acid. Since tissue sites were reduced by chondroitinase but not heparitinase, we suggest that degradation is due to hyaluronidase activity of chondroitinase and the anionic sites are associated with hyaluronic acid. However, the influence of pH indicates that lamina rara externa sites are structurally distinct from other GBM anionic sites.

scholarly journals Cationic colloidal gold--a new probe for the detection of anionic cell surface sites by electron microscopy.

1986 ◽  
Vol 34 (5) ◽  
pp. 693-696 ◽  
Author(s):  
E Skutelsky ◽  
J Roth
Keyword(s):  
Cell Surface ◽  
Colloidal Gold ◽  
Particle Diameter ◽  
Neuraminidase Treatment ◽  
Surface Sites ◽  
Ph Values ◽  
Red Blood Cell Membranes ◽  
Cationic Colloidal Gold ◽  
Flexible Probe ◽  
Cellular Components

Particles of colloidal gold were coated with poly-L-lysine to prepare cationic colloidal gold. Monodispersed colloidal gold with a particle diameter of 5, 8, or 15 nm and poly-L-lysine with a molecular weight of 350,000 or 1500-8000 were used. The resulting complexes were used to label red blood cell membranes. The labeling was sensitive to neuraminidase treatment or acid hydrolysis, demonstrating that cationic colloidal gold binds preferentially to anionic cell surface constituents. Cationic colloidal gold can be used at physiological pH values and ionic strength, as well as at low pH values, making it a flexible probe for detection of anionic cellular components.

scholarly journals Sulfated Glycosaminoglycans In Guinea Pig Neutrophils Studied by Use of Cationic Colloidal Gold

1999 ◽  
Vol 47 (7) ◽  
pp. 881-887 ◽  
Author(s):  
Dong-Hua Yang ◽  
Shinichiro Tsuyama ◽  
Jun Ohmori ◽  
Fusayoshi Murata
Keyword(s):  
Guinea Pig ◽  
Colloidal Gold ◽  
Cationic Colloidal Gold

Formation of basement membrane-like structure terminates the cellular encapsulation of microfilariae in the haemocoel of Anopheles quadrimaculatus

Parasitology ◽  
1998 ◽  
Vol 116 (6) ◽  
pp. 511-518 ◽  
Author(s):  
C. T. LIU ◽  
R. F. HOU ◽  
C. C. CHEN
Keyword(s):  
Basement Membrane ◽  
Colloidal Gold ◽  
Early Stage ◽  
Basement Membranes ◽  
Brugia Pahangi ◽  
Exterior Surface ◽  
Anopheles Quadrimaculatus ◽  
Outermost Surface ◽  
Cationic Colloidal Gold

The encapsulation of microfilariae in the haemocoels of mosquitoes combines both humoral and cellular reactions: the microfilariae are first encased in an acellular layer of melanin, followed by a cellular encapsulation by plasmatocytes. In this study, we demonstrated that cellular encapsulation of Brugia pahangi microfilariae in the haemocoel of the mosquito Anopheles quadrimaculatus was terminated by the formation of a basement membrane-like structure on the outermost surface of the cellular capsule. This structure occurred in the early stage of cellular encapsulation and was evident on the exterior surface of the plasmatocyte, when the active haemocytes were attaching to the already melanized microfilariae. The termination structure appears to be laid down by releasing the vesicle inclusions of haemocytes and has similarities in ultrastructure and cationic colloidal gold staining properties with that of mosquito basement membranes.

Distribution of lectin binding sties and anionic sites in the rat zona pellucida

1990 ◽  
Vol 48 (3) ◽  
pp. 64-65
Author(s):  
E. Skutelsky ◽  
R. Maymon ◽  
B. Bar-Shira (Maymon) ◽  
R. Shalgi
Keyword(s):  
Zona Pellucida ◽  
Colloidal Gold ◽  
Lectin Binding ◽  
Cumulus Cells ◽  
Ultrastructural Level ◽  
Female Rats ◽  
Anionic Sites ◽  
Specific Nature ◽  
Immature Female ◽  
Specific Sugar

The zona pellucida (ZP) in mammals is an extracellular coat which surroudns the oocyte during growth, fertilization and early development, consisting of a limited number of glycoproteins. Carbohydrates residues and charge properties of the ZP are believed to have a role in sperm-egg interaction. The purpose of the present study was to furtehr characterize the specific nature and distribution of the carbohydrate residues of the ZP and cumulus cells and anionic residues at the ultrastructural level.We used lectins as probes for specific sugar residues and polycationic colloidal gold (PCG) as a marker for aninonic sites. Immature female rats were induced to superovate. Ovulated, cumulus-oocyte complexes were isolated and fixed with 2.5% Glutaraldehyde, dehydrated and embedded in Araldite or LR-Gold. For light microscopic (LM) histochemistry, 0.5 μm sections were cut, deresined by Na-methoxide and labeled with biotin-derivatized lectins as probes and avidin biotin-peroxidase complex as visualant.

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