Special stains used for connective tissues

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					Special Stains of Connective Tissues
Dr. Muhammad Rafique

Hypothesis
What is a stain Role of ordinary stains Ordinary satins Why the special stains used What are the examples of special stains for Connective tissues

Staining and visualisation
After the new sections have been baked onto their slides it is necessary to stain them in order that they can be viewed under the microscope. Without staining, the view is of grey crystalline material - this is paraffin wax. The most common stain used throughout the world as the basic morphology preparation is the Haematoxylin and Eosin stain. Haematoxylin is a natural dye extracted from trees. It is a basic dye and therefore binds with acidic tissue components. Eosin on the other hand, is an acidic dye and binds with basic tissue components including the cell membranes and organelles.

Used of Special Stains
Special stains are used to demonstrate the different components of tissues which are stained with different colors Special stains usually consist of combinations different stains so that different components of tissues take up different stain. For e.g. collagen fibers, muscle fibers, elastic fibers and reticular fibers take different colors and identified separately

Special Stains Used for Connective Tissues
• • • • Masson's Trichrome Stain for CT Mallory's Connective Tissue Stain Verhoeff-van Gieson Stain Masson Trichrome & Verhoeff Stain

Masson's Trichrome Stain for CT
This is an example of Masson's Trichrome Stain for CT. There are many variations of this routine. In most of them collagenous components are stained a green or blue color and cytoplasm is varying shades of red. In this it's easy to make out the extensive collagenous fiber reinforcement of the wall of the large vein at left, and the CT wrappings of the bundles of nerve fibers at right. The higher magnification image shows the CT of the vein wall in detail.

Masson's Trichrome Stain for CT
Ordinary Stain Special Stain

Mallory's Connective Tissue Stain
The example here is from a section of the pancreas. The CT stroma (supporting elements) of this organ are highlighted in blue using this stain, and the parenchymal tissue (i.e., the functional secretory cells) is stained in red. Mallory's CT stain is very widely used and many variations on it exist.

Mallory's Connective Tissue Stain
Ordinary Stain

Special Stain

Verhoeff-van Gieson Stain
Verhoeff component of this two part stain is specific for elastic fibers. The Van Gieson stain is specific for collagen. This combination permits to distinguish two types of CT elements in the wall of this artery: the black strands are elastic fibers and the bright red material is the reinforcing collagen. In addition to these CT elements, there's a lot of smooth muscle in the wall of this artery, which is stained a duller red than the bright crimson of the collagen.

Verhoeff-van Gieson Stain
Ordinary Satin

Special Stain

Masson Trichrome & Verhoeff Stain
In it the Masson's Trichrome stain, and the Verhoeff stain, both routines for connective tissue, have been combined. The Masson stain colors collagen green and muscle red; the Verhoeff stain colors elastic fibers black. Using a combination method like this one makes it possible to provide a lot of detail about how an organ is built. Black elastic fibers are interspersed throughout the muscle to provide springiness and resilience; the green collagen fibers confer resistance to overstretch.

Masson Trichrome & Verhoeff Stain
Ordinary Stain Special Satin

MASSON’S TRICHROME STAIN
Purpose Trichrome stains are frequently used to differentiate between collagen and smooth muscle in tumors and to identify increases in collagenous tissue in diseases such as cirrhosis of the liver. Components Solution A Hematoxylin ... 10.0 g Alcohol, 95% ... 1,000.0 ml Solution B Ferric chloride, 29% aqueous solution ... 20.0 ml Distilled water ... 475.0 ml Glacial acetic acid ... 5.0 ml Working Solution Mix equal parts of solutions A and B. Results Nuclei ..... black Cytoplasm, keratin, muscle fibers ..... red Collagen and mucus ..... blue

Mallory's Trichrome for Muscle and Collagen
Solutions Solution A Acid fuchsin 1 g Distilled water 100 mL Solution B Phosphomolybdic acid 1 g Distilled water 100 mL Solution C Orange G 2 g Methyl blue 0.5 g Oxalic acid 2 g Distilled water 100 mL Expected results Nuclei – red Erythrocytes – orange Muscle – red Collagen – blue