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Connective & Muscle
Connective and muscle tissue in the histology lab
| Question | Answer |
|---|---|
| Three types of connective tissue | 1. Connective tissue proper 2. Cartilage 3. Bone |
| Connective tissue proper breaks down into three categories | 1. Collagen 2. Elastic 3. Reticulin |
| Collagen | Makes tissue strong |
| Dense regular collagen | Tendons, organ capsules, and the dermis |
| Elastic tissue | Found in tissue requiring flexibility |
| Reticulin | Delicate supporting network for highly cellular organs such as endocrine glands, lymph nodes, and liver |
| Basement membrane | Basal lamina |
| Three different type of muscle | 1. skeletal 2. cardiac 3. smooth |
| Skeletal muscle | Voluntary, striated |
| Cardiac muscle | Involuntary, striated |
| Smooth muscle | Involuntary, non striated |
| Masson Trichrome | Used to differentiate between collagen and smooth muscle in tumors to identify increases in collagenous tissue in diseases such as cirrhosis of the liver |
| Masson Trichrome | Bouin, 10% NBF is ok |
| Masson Trichrome | 4-5 micrometers |
| Masson Trichrome control | Every tissue has an internal control but uterus, small intestine, appendix, or Fallopian tubes |
| Masson Trichrome | 1. Bouin 2. Rinse 3. Weigert Iron 4. rinse 5. Biebrich scarlet acid fuchsin 6. rinse 7. Place in phosphomolybdic. phosphotungstic acid 8. Aniline blue 9. Rinse 10. 1% Acetic Acid 11. Dehydrate 12. Clear |
| Masson Trichrome results | Nuclei: Black Cytoplasm, keratin, muscle fibers: Red Collagen and mucin: Blue |
| When collagen is predominant then this counterstain is preferred | Light green |
| Masson Trichrome results | Nuclei: Dark blue Cytoplasm, keratin, muscle fibers: Red Collagen and mucin: Blue |
| Gomori 1-step Trichrome | Identify increase in collagenous connective tissue fibers or to differentiate between collagen and smooth muscle fibers |
| Gomori 1-step Trichrome | Chromotrope 2 R and fast green FCF, light green or aniline blue are combined in a solution of phosphotungstic acid to which glacial acetic acid has been added. |
| Gomori 1-step Trichrome | Bouin |
| Gomori 1-step Trichrome | 4-5 micrometers |
| Gomori 1-step Trichrome control | All tissue have internal controls |
| Gomori 1-step Trichrome | 1. Bouin 2. Rinse 3. Weigert iron heme 4. wash 5. Gomori Trichrome 6. Differentie with acetic acid .5% 7. Dehydrate 8. Clear |
| Gomori 1-step Trichrome results | Nuclei: Black Cytoplasm, keratin, muscle fibers: Red Collagen and mucins: Green or blue |
| Van Gieson picric acid acid fuchsin stain | Stains for collagen |
| Van Gieson picric acid acid fuchsin stain | Any type of fixative can be used |
| Van Gieson picric acid acid fuchsin stain | 4-5 micrometers |
| Van Gieson picric acid acid fuchsin stain control | Almost all tissue has an internal control. Uterus, appendix, small intestine or Fallopian tube is a good control |
| Van Gieson picric acid acid fuchsin stain | 1. Weigert iron heme 2. rinse 3. Van Gieson stain 5. Dehydrate 6. Clear |
| Van Gieson picric acid acid fuchsin stain results | Nuclei: Black Collagen: Red Muscle and cytoplasm: Yellow |
| Verhoeff Elastic | Used to demonstrate pathological changes in elastic tissue |
| Verhoeff Elastic | Ferric Chloride and iodine serve as mordants |
| Verhoeff Elastic | 10% NBF or Zenker preferred |
| Verhoeff Elastic | 4-5 micrometers |
| Verhoeff Elastic control | Aorta |
| Verhoeff Elastic | 1. Verhoeff stain 2. rinse 3. differentiate in 2% ferric chloride 4. rinse 5. place in sodium thiosulfate 6. wash 7. counterstain 8. differentiate 9. dehydrate 10. clear |
| Verhoeff Elastic results | Elastic fibers: Black Nuclei: Blue to black Collagen: Red Other tissue elements: Yellow |
| If an elastic stain is over differentiated it can be restained provided it has not be treated with what? | Alcohol |
| Aldehyde fuchsin elastic | Used to stain highly sulfated mucosubstances, pancreatic beta cells, and hepatitis b antigen |
| Aldehyde fuchsin elastic | 10% NBF, chromate fixatives should be avoided (Shows a lilac background) |
| Aldehyde fuchsin elastic | 4-5 micrometers |
| Aldehyde fuchsin elastic control | Aorta or muscular artery |
| Aldehyde fuchsin elastic | 1. Aldehyde fuchsin 2. rinse with 70% alcohol 3. wash 4. counter stain with light green 5. dehydrate 6. clear |
| Aldehyde fuchsin elastic results | Elastic: Deep blue to purple Other tissue elements: Green |
| Russell modification of the Movat pentachorme stain | Demonstration of mucin, fibrin, elastic fibers, muscle and collagen |
| Russell modification of the Movat pentachorme stain | 10% NBF |
| Russell modification of the Movat pentachorme stain | 4-5 micrometers |
| Russell modification of the Movat pentachorme control | Lung, skin or colon |
| Russell modification of the Movat pentachorme results | Nuclei and elastic fibers: black Collagen: Yellow Ground substances and mucin: blue Fibrinoid, fibrin: Intense red Muscle: Red |
| Oxidizers | Phosphomolybdic acid, potassium permanganate, and periodic acid |
| Sensitization | Uranyl nitrate, ferric ammonium sulfate, and dilute solutions of silver nitrate |
| Silver impregnation | Ammonical or diamine silver complex |
| Reduction | Formaldehyde |
| Toning | Gold chloride |
| Unreduced silver is removed by | Sodium thiosulfate |
| Gomori stain for reticular fibers | Demonstration of reticular fibers in tissue |
| Gomori stain for reticular fibers | Important in the differential diagnosis of certain types of tumors. |
| Gomori stain for reticular fibers | 10% NBF |
| Gomori stain for reticular fibers | 4-5 micrometers |
| Gomori stain for reticular fibers control | Liver |
| Gomori stain for reticular fibers | 1. Potassium Permanganate 2. rinse 3. potassium meta bilsulfite 4. wash 5. 2% Ferric ammonium sulfite 6.wash 7. Ammonical silver 8. rinse 9. 20% formalin 10. wash 11. Gold Chloride 12. rinse 13. Potassium metabilsulfite 14. W 15. Counterstain |
| Gomori stain for reticular fibers results | Reticulin: Black Collagen: Taupe |
| Gordon & Sweets for Reticulin | Demonstration of reticular fibers |
| Gordon & Sweets for Reticulin | 10% NBF |
| Gordon & Sweets for Reticulin | 4-5 micrometers |
| Gordon & Sweets for Reticulin control | Liver |
| Gordon & Sweets for Reticulin | 1. Potassium Permanganate 2. Rinse 3. Bleach in 1% oxalic acid 4. Wash 5. 2.5% Ferric Ammonium sulfate 6. Wash 7. Silver solution 8. 10% Formalin 9. Wash 10. Gold Chloride 11. Wash 12. 5% sodium thiosulfate 13. Rinse 14 Counterstain |
| Gordon & Sweets for Reticulin results | Reticulin: Black Other tissue elements depend on counter stain used |
| Mallory PTAH technique for cross striations & fibrin | Demonstration of muscle cross striations and fibrin |
| Mallory PTAH technique for cross striations & fibrin | Diagnostic of rhabdomyosarcomas or tumors arising from striated muscles |
| Mallory PTAH technique for cross striations & fibrin | Zenker preferred, 10% NBF may be used |
| Mallory PTAH technique for cross striations & fibrin | 4-6 micrometers |
| Mallory PTAH technique for cross striations & fibrin control | Longitudinal sections of skeletal or cardiac muscle to demonstrate cross striations, section containing fibrin for the demonstration of fibrin |
| Mallory PTAH technique for cross striations & fibrin | 1. Mordant in Zenker 2. rinse 3. Gram iodine 4. rinse 5. 5% sodium thiosulfate 6. wash 7. Potassium permanganate 8. rinse 9. oxalic acid 10. wash 11. stain PTAH solution 12. Dehydrate 13. Clear |
| Mallory PTAH technique for cross striations & fibrin results | Cross striations, fibrin: Blue Nuclei: Blue Collagen: Red-brown |
| Periodic Acid methnamine silver microwave procedure for basement membranes | 2 micrometers |
| Periodic Acid methnamine silver microwave procedure for basement membranes control | kidney has an internal control built in |
| Periodic Acid methnamine silver microwave procedure for basement membranes | 1. Periodic acid 2. Rinse 3. Methnamine working solution 4. rinse 5. gold chloride 6. rinse 7. Sodium thiosulfate 8. wash 9. counterstain 10. dehydrate 11. Clear |
| Periodic Acid methnamine silver microwave procedure for basement membranes results | Basement membrane: Black Background: green |
| Oil Red O | Demonstration of neutral lipids |
| Oil Red O | Physical method |
| Oil Red O | 10% NBF or calcium formalin |
| Alcohol will do what to fat | Dissolve lipids |
| Oil Red O | Cut frozen at 10 micrometers |
| Oil Red O control | Most tissue contains some fat, so normally a control is not used |
| Oil Red O | 1. stain in oil red 2. rinse 3. stain in Harris Heme with acetic acid 4. wash 5. Blue in ammonia water 6. wash 7. mount |
| Oil Red O results | Fat: red Other tissue elements: according to method used |
| Sudan Black | Demonstration of neutral lipids |
| Sudan Black | Most sensitive of lipid dyes |
| Sudan Black | 10% NBF or calcium formalin |
| Sudan Black | No alcohol should be used at all |
| Sudan Black | 10 micrometers |
| Sudan Black control | Most tissue contains fat |
| Sudan Black | 1. Sudan black 2. Differentiate with propylene glycol 3. wash 4. Counterstain 5. wash 6. mount |
| Sudan Black results | Fat: Blue-black Nuclei: red |
| Osmium Tetroxide | Demonstration of fat and allows for paraffin embedding of tissue |
| Osmium Tetroxide | Chemically combines with fat, blackening it in the process |
| Osmium Tetroxide | 10% NBF |
| Osmium Tetroxide control | No control neccesary |
| Osmium Tetroxide results | Fat: Black |
| Toluidine blue for Mast cells | Demonstration of mast cells in tissue |
| Toluidine blue for Mast cells | 10% NBF |
| Toluidine blue for Mast cells | 4-5 micrometers |
| Toluidine blue for Mast cells control | Section containing mast cells |
| Toluidine blue for Mast cells | 1. Toluidine blue 2. rinse 3. dehydrate 4. clear |
| Toluidine blue for Mast cells results | Mast cells: Deep rose violet Background: Blue |
Created by:
Ziek98
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