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Connective & Muscle

Connective and muscle tissue in the histology lab

QuestionAnswer
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