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Connective Tissue
Special Stain for demonstration of Connective Tissue
| Question | Answer |
|---|---|
| What is the purpose of Masson's Trichrome? | To demonstrate between collagen and smooth muscle in tumors and the increase of collagen in diseases as cirrhosis. |
| What is a disease that shoes increase of collagen? | Cirrhosis |
| What is a routine stain for liver and kidney biopsies? | Masson's Trichrome |
| State the principle of the Masson's Trichrome | Three dyes are employed. The general rule in trichrome staining is that the less porous tissues are colored by the smallest dye molecules, whenever a dye of larger molecular size is able to penetrate, it will do so at the expense of the smaller molecule. |
| Which elements does Masson's Trichrome stain specifically? | Muscle, collagen, fibrin and erythrocytes |
| Why do we stain tissue with Wiegerts Haematoxylin? | To stain the entire nuclei |
| What is the differentiating reagent in Masson's Trichrome? | Acid Alcohol |
| What is Scott's Tap Water used for in Masson's Trichrome? | The blueing step |
| What is the cytoplasm stain for Masson's Trichrome? | Ponceau |
| What is Ponceau used for in Masson's Trichrome? | To stain the cytoplasm and other elements |
| What is the the mordant and differentiation reagent in Masson's Trichrome? | Phosphomolybdic Acid |
| How long do you treat sections with phosphomolybdic acid? | Until collagen is decolourised and muscle, red blood cells and fibrin remains red |
| How do you monitor the phosphomolybdic acid Reaction in Masson's Trichrome? | Under the microscope |
| What counterstain is used in Masson's Trichrome? | Light Green |
| With what reagent do you differentiate the counterstain in Masson's Trichrome? | Acetic Acid |
| What colour will the nuclei be stained in Masson's Trichrome? | Blue/Black |
| What colour will the cytoplasm, muscle, acidophil granules, RBCs and fibrin stain in Masson's Trichrome? | Red |
| What colour will collagen, cartilage, mucin and basophil granules stain in Masson's Trichrome? | Green |
| What is the control tissue used in Masson's Trichrome? | Uterus, appendix or fallopian tubes |
| State the purpose of Gomori's One Step Trichrome | To identify an increase in collagenous tissue fibers or to differentiate between collagen and smooth muscle fibres |
| What is the principle of Gomori's One Step Trichrome? | Based on a single step staining. In this procedure a plasma stain (Chromophore 2R) and a connective tissue stain (fast green FCF, light green or aniline blue) are combined in a solution of phosphotungstic acid to which glacial acetic acid has been added |
| What is the connective tissue stain in the Gomori's One Step? | Fast Green FCF, light green or aniline blue |
| What is the plasma stain in Gomori's One Step? | Chromophore 2R |
| What does the combined solution in Gomori's One Step consists of? | Phosphotungstic acid and glacial acetic acid |
| Why do you use Weigert's Haematoxylin in in Gomori's One Step? | To stain the nuclei |
| What is the nuclei stained with in Gomori's One Step? | Wiegerts Haematoxylin |
| Why do you stain with Gomori's Trichrome solution? | To demonstrate the collagen and smooth muscle |
| What colour will the nuclei be stained in the Gomori's One Step? | Black |
| What colour will the cytoplasm, keratin and muscle fibres be stained in the Gomori's One Step? | Red |
| What colour will the collagen and mucous be stained in the Gomori's One Step? | Green/Blue (Counterstain depending) |
| What control tissue is used in Gomori's One Step? | Uterus, appendix or fallopian tubes |
| State the purpose and principle of the Van Gieson stain? | Van Gieson is used to differentiate between collagen and smooth muscle in tumors and to demonstrate the increase of collagen in diseases |
| What is used to stain the nuclei detail in Van Gieson? | Wiegerts Haematoxylin |
| What type of Haematoxylin is Wiegerts? | Iron Heamatoxylin |
| Why is it important to wash sections after staining with Wiegerts Haematoxylin? | To remove excess Haematoxylin |
| What is the counterstain in Van Gieson? | Van Gieson |
| Why do we rapidly dehydrated sections through alcohols? | Certain Stains can be removed from the sections |
| What colour will the nuclei be stained in the Van Gieson? | Red |
| What colour will the nuclei be stained? | Dark blue/Black |
| What colour will the muscle and RBC be stained in the Van Gieson? | Yellow |
| What control tissue is used in Van Gieson? | Uterus, appendix or fallopian tubes |
| Name three stains used to demonstrate muscle and collagen | Masson's Trichrome Gomori's One Step Trichrome Van Gieson |
| Name two trichrome staining method s | Masson's Trichrome Gomori's One Step Trichrome |
| What is the difference between the Masson's Trichrome and Gomori's One Step Trichrome methods? | Masson's Trichrome uses three dyes to stain elements specifically where as the Gomori's One Step only one dye is used to stain different colours other than the dye colour itself |
| Which fixative is best for post fixation in trichrome staining? | Bouin's |
| Why is Bouin's fixative not the routine fixative in trichrome staining? | It can produce pigments and artifacts |
| State the purpose of the EVG method? | Demonstration of pathological changes in elastic fibres in cases of emphysema and/or arteriosclerosis |
| What diseases are associated with with the Verhoeff's Van Gieson? | Emphysema and arteriosclerosis |
| State the principle of the EVG | The tissue is overstrained with Verhoeff's Haematoxylin . Ferric chloride and Iodine serves as the mordant and oxidizing agent, converting the Haematoxylin to haematin. The ferric chloride will differentiate the elastic fibres until they are clearly seen. |
| What are the components of the Verhoeff's Haematoxylin? | Alcoholic Haematoxylin Ferric chloride Lugol's Iodine |
| Why do you prepare the elastic stain fresh every time before use in the EVG? | To avoid poor staining results |
| How long do you differentiate in the EVG? | Until the elastic fibres stand out sharply and the background is clear |
| How do you monitor the differentiation progress in the EVG? | Under the microscope |
| What is the differentiating in the EVG? | Ferric chloride |
| State the counterstain used in the EVG | Van Gieson |
| What colour will collagen be stained in the EVG? | Red |
| What colour will the nuclei be stained in the EVG? | Dark blue |
| What colour will the background be stained in the EVG? | Yellow |
| What colour will the elastic fibres stain in the EVG? | Black |
| What control tissue is used in the EVG? | Aorta or large Artery |
| What is the full name of the EVG? | Verhoeff's Van Gieson |
| What is the full name for PHAH? | Phosphotungstic Acid Haematoxylin |
| State the purpose of the PTAH stain | Used for demonstrating muscle cross striations, fibrin and glial fibers |
| State the principle of PTAH | The stain has been referred to as a polychromatic stain because one solution gives two colours. The components coloured red-brown will lose this colour with prolonged alcohol washes, and dehydration of the section thereafter must be rapid. |
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