scholarly journals SUGGESTIONS FOR A HISTOCHEMICAL TERMINOLOGY OF CARBOHYDRATE-RICH TISSUE COMPONENTS

1965 ◽  
Vol 13 (7) ◽  
pp. 599-603 ◽  
Author(s):  
S. S. SPICER ◽  
T. J. LEPPI ◽  
P. J. STOWARD
Keyword(s):  
Sialic Acid ◽  
Connective Tissue ◽  
Acid Mucopolysaccharide ◽  
Tissue Sections ◽  
Histochemical Methods ◽  
General Terms

Biochemically known mucopolysaccharides can rarely be identified unequivocally in specific tisse sites by histochemical methods. Moreover, many carbohydrate-rich substances that have been detected histochemically (in terms of their vic-glycol content and characteristic acid groups) cannot be related to mucopolysaccharides and glycoproteins known to biochemists. Accordingly, a terminology and classification is proposed for delineating mucosubstances in situ in terms of their histochemical properties. Mucosubstance and mucosaccharide are recommended as general terms for tissue entities rich in carbohydrate, but the term [acid] mucopolysaccharide is restricted to biochemically known mucopolysaccharides of connective tissue. The terms sulfomucin and sialomucin are suggested for acidic epithelial mucosubstances containing sulfate esters and sialic acid, respectively. Neutral mucosubstance is reserved for naming periodate-reactive entities, except glycogen, that lack demonstrable acid groups. The various muscosubstances that have been found in tissue sections so far are tabulated with suggested names.

Flat embeddment of monolayer cells, tissue sections, and other cellular materials attached onto glass substrate

1990 ◽  
Vol 48 (3) ◽  
pp. 766-767
Author(s):  
Jeffrey P. Chang ◽  
Jaang J. Wang
Keyword(s):  
Present Report ◽  
Cellular Materials ◽  
Cover Glass ◽  
Tissue Sections ◽  
Embedding Technique ◽  
Exfoliated Cells ◽  
Black Paper ◽  
Cell Concentrations ◽  
Flat Embedding

Flat embeddment of certain specimens for electron microscopy is necessary for three classes of biological materials: namely monolayer cells, tissue sections of paraffin or plastics, as well as cell concentrations, exfoliated cells, and cell smears. The present report concerns a flat-embedding technique which can be applied to all these three classes of materials and which is a modified and improved version of Chang's original methodology.Preparation of coverglasses and microslides. Chemically cleaned coverglasses, 11 × 22 mm or other sizes, are laid in rows on black paper. Ink-mark one coner for identifying the spray-side of the glass for growing cells. Lightly spray with Teflon monomer (Heddy/Contact Inductries, Paterson, NO 07524, U.S.A.) from a pressurized can. Bake the sprayed glasses at 500°F for 45 min on Cover-Glass Ceramic Racks (A. Thomas Co. Philadelphia), for Teflon to polymerize.Monolayer Cells. After sterilization, the Teflon-treated coverglasses, with cells attached, are treated or fixed in situ in Columbia staining dishes (A. Thomas Co., Philadelphia) for subsequent processing.

Porcine Repeat Element DNA: In Situ Detection of Xenotransplanted Cells

1995 ◽  
Vol 4 (2) ◽  
pp. 253-256 ◽  
Author(s):  
Henry F. Oettinger ◽  
Amelie Rodrigue-Way ◽  
Joyce J. Bousquet ◽  
Albert S.B. Edge
Keyword(s):  
Southern Blotting ◽  
Host Tissue ◽  
Repeat Element ◽  
Tissue Analysis ◽  
Specific Staining ◽  
Tetrazolium Chloride ◽  
Tissue Sections ◽  
Blue Tetrazolium Chloride ◽  
In Situ Detection

Using a digoxygenin-labelled DNA probe derived from the porcine repeat element PRE-1, we have developed a protocol for the detection of transplanted porcine islets and hepatocytes against a background of murine host tissue. Analysis of this probe by Southern blotting indicated that PRE-1 hybridizes to pig genomic DNA but not to human or mouse DNA. On tissue sections, hybridizing probe was detected using alkaline phosphatase-conjugated anti-digoxygenin antibody visualized with 5-bromo-4-chloro-3-indolyl-phosphate/4-nitro-blue tetrazolium chloride (BCIP/ NBT) substrate. We have demonstrated sensitive and highly specific staining of porcine nuclei in fixed, paraffin embedded tissue sections, and have applied the technique to detect porcine pancreatic islets and hepatocytes transplanted into murine kidney and spleen. Applications of this technique include detection of transplanted cells or organs across a variety of xenogeneic barriers.

Molecular Hybridization of Mouse Satellite DNA-Complementary RNA in Ultrathin Sections Prepared for Electron Microscopy

1974 ◽  
Vol 14 (2) ◽  
pp. 253-261
Author(s):  
J. JACOB ◽  
KATHERINE GILLIES ◽  
D. MACLEOD ◽  
K. W. JONES
Keyword(s):  
Electron Microscopy ◽  
Satellite Dna ◽  
Similar Distribution ◽  
Tissue Sections ◽  
Satellite Sequences ◽  
Interphase Cells ◽  
Nucleolar Chromatin ◽  
Ultrathin Sections ◽  
Mouse Satellite Dna

The feasibility of in situ hybridization in tissue sections prepared for electron microscopy has been examined using mouse satellite DNA-complementary RNA and mouse L cells. The results obtained are encouraging, although certain technical aspects require further clarification. In interphase cells, hybrid-forming sites occur in chromatin patches positioned along the nuclear envelope. It is also confirmed that satellite DNA occurs in nucleolus-associated chromatin. The results suggest that satellite sequences are present in intranucleolar and peri-nucleolar chromatin. A similar distribution is indicated for ribosomal cistrons.

scholarly journals A new method to detect apoptosis in paraffin sections: in situ end-labeling of fragmented DNA.

1993 ◽  
Vol 41 (1) ◽  
pp. 7-12 ◽  
Author(s):  
J H Wijsman ◽  
R R Jonker ◽  
R Keijzer ◽  
C J van de Velde ◽  
C J Cornelisse ◽  
...  
Keyword(s):  
Cell Death ◽  
Programmed Cell Death ◽  
Staining Method ◽  
Morphological Characteristics ◽  
Image Cytometry ◽  
Apoptotic Cells ◽  
Dna Breaks ◽  
Paraffin Sections ◽  
Tissue Sections

Apoptosis (programmed cell death) can be difficult to detect in routine histological sections. Since extensive DNA fragmentation is an important characteristic of this process, visualization of DNA breaks could greatly facilitate the identification of apoptotic cells. We describe a new staining method for formalin-fixed, paraffin-embedded tissue sections that involves an in situ end-labeling (ISEL) procedure. After protease treatment to permeate the tissue sections, biotinylated nucleotides are in situ incorporated into DNA breaks by polymerase and subsequently stained with DAB via peroxidase-conjugated avidin. Staining of cells with the morphological characteristics of apoptosis was demonstrated in tissues known to exhibit programmed cell death, i.e., prostate and uterus after castration, tumors, lymph node follicles, and embryos. Apoptotic cells could be discriminated morphologically from areas of labeled necrotic cells, in which DNA degradation also occurs. Because apoptosis is relatively easily recognized in H&E-stained sections of involuting prostates of castrated rats, we used this model system to validate the ISEL method for the quantification of apoptotic cells. A high correlation was found between the fractions of ISEL-labeled cells and the fractions of apoptotic cells that were morphologically determined in adjacent sections. We conclude that ISEL is a useful technique for quantification of apoptosis in paraffin sections, especially for those tissues in which morphological determination is difficult. Furthermore, this new staining method enables the use of automated image cytometry for evaluating apoptosis.

scholarly journals Exo-enzymatic addition of diazirine-modified sialic acid to cell surfaces enables photocrosslinking of glycoproteins

2021 ◽  
Author(s):  
Nageswari Yarravarapu ◽  
Rohit Sai Reddy Konada ◽  
Narek Darabedian ◽  
Nichole J. Pedowtiz ◽  
Soumya N. Krishnamurthy ◽  
...  
Keyword(s):  
Sialic Acid ◽  
Cell Surface ◽  
Cell Types ◽  
Cell Surfaces ◽  
Binding Interactions ◽  
Multiple Cell ◽  
Labeling Method ◽  
Cell Surface Glycoconjugates

Glycan binding often mediates extracellular macromolecular recognition events. Accurate characterization of these binding interactions can be difficult because of dissociation and scrambling that occur during purification and analysis steps. Use of photocrosslinking methods has been pursued to covalently capture glycan-dependent interactions in situ however use of metabolic glycan engineering methods to incorporate photocrosslinking sugar analogs is limited to certain cell types. Here we report an exo-enzymatic labeling method to add a diazirine-modified sialic acid (SiaDAz) to cell surface glycoconjugates. The method involves chemoenzymatic synthesis of diazirine-modified CMP-sialic acid (CMP-SiaDAz), followed by sialyltransferase-catalyzed addition of SiaDAz to desialylated cell surfaces. Cell surface SiaDAz-ylation is compatible with multiple cell types and is facilitated by endogenous extracellular sialyltransferase activity present in Daudi B cells. This method for extracellular addition of α2-6-linked SiaDAz enables UV-induced crosslinking of CD22, demonstrating the utility for covalent capture of glycan-mediated binding interactions.

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