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Nerve

Nerve stains in the histology lab

QuestionAnswer
What are the two parts of the nervous system? Central and peripheral
CNS Spinal cord and brain
Somatic nervous system Voluntary (conscious control)
Autonomic nervous system Involuntary
Four types of glial cells 1. Oligodendroglia 2. Astrocytes 3. Microglia 4. Ependymal cells
Myelin consists of protein, chloesterol, phospholipids, and cerebrosides
Crsyl echt violet method I To identify neurons in tissue sections or the demonstration of the loss of Nissl substance
Crsyl echt violet method I Neurons contain nissl substance, which is primarily composed of rough endoplasmic reticulum. Nissl substance is basophilic and stains well with aniline dyes.
Crsyl echt violet method I 10% NBF
Crsyl echt violet method I 6-8 micrometers
Crsyl echt violet method I control Spinal cord
Crsyl echt violet method I 1.Crsyl violet 2. Rinse 3. 95% alcohol 4. 100% alcohol 5. xylene 6. balsam-xlene 7. 100% alcohol 8. Xylene (Steps 6-8 will have to be repeated till proper differentiation is reached)
Crsyl echt violet method I results Nissl substance: Blue to purple Nuclei: Blue to purple Background: Colorless
Creysl echt violet method II Identify neurons in tissue sections or the demonstration of loss of Nissl substance (chromatolysis)
Creysl echt violet method II 10% NBF
Creysl echt violet method II 6-8 micrometers
Creysl echt violet method II control spinal cord
Creysl echt violet method II 1. Cresyl echt violet 2. Dehydrate 3. Clear
Creysl echt violet method II Nissl substance and nuclei: Blue-purple Background: Colorless
Bodian method Nerve fibers, nerve endings, neurofibrils
Wallerian degeneration When an axon is severely or irreversibly injured, all of the axon distal to the injury disappears along with its myelin sheath
Bodian method 10% NBF
Bodian method 6-8 micrometers
Bodian method control Peripheral nerve or cerebral cortex
Bodian method 1. Protargol 2. rinse 3. reducing solution 4. rinse 5. gold chloride 6. rinse 7. Oxalic acid 8. rinse 9. sodium thiosulfate 10. rinse 11. counterstain with aniline blue 12. dehydrate 13. clear
Bodian method results Nerve fibers: Black Background: Light gray or blue Nuclei: Black
Holmes silver nitrate method Demonstration of Nerve fibers and neurofibrils
Holmes silver nitrate method 10% NBF
Holmes silver nitrate method 10-15 micrometers
Holmes silver nitrate method control Cerebral cortex
Holmes silver nitrate method 1. 20% silver nitrate 2. rinse 3. impregnating solution 4. reduce 5. wash 6. gold chloride 7. rinse 8. 2% Oxalic acid 9. rinse 10. 5% sodium thiosulfate 11. wash 12. dehydrate 13. clear
Holmes silver nitrate method results Axons and nerve fibers: Black Neurofibrils: Black
Bielschowsky-PAS Nerve fibers, neurofibrillary tangles, and senile plaques
Bielschowsky-PAS Diagnoses Alzheimer
Bielschowsky-PAS 10% NBF
Bielschowsky-PAS 8-10 micrometers
Bielschowsky-PAS control CNS that contains senile plaques and neurofibrillary tangles
Bielschowsky-PAS 1. 20% silver nitrate 2. w 3. Ammonical silver solutions 4. w in ammonia water 5. Add developer 6. w 7. gold chloride 8. Ammonia water 9. 5% sodium thiosulfate 10. w 11. 1% periodic acid 12. wash 13. Schiff 14. wash 15. Dehydrate and clear
Bielschowsky-PAS results Neurofibrillary tangles and peripheral neuritis of neuritic plaques: Dark black Axons: Black Amyloid(plaque cores and vascular): Magenta Lipofuscin: Magenta
Seiver-Munger modification of Bielschowsky Nerve fibers, neurofibrillary tangles and senile plaques
Seiver-Munger modification of Bielschowsky Diagnoses Alzheimers
Seiver-Munger modification of Bielschowsky 6-8 micrometers
Seiver-Munger modification of Bielschowsky control CNS
Seiver-Munger modification of Bielschowsky 1. 20% silver nitrate 2. rinse 3. Formalin 4. rinse 5, Sodium thiosulfate 6. wash 7. Dehydrate and clear
Seiver-Munger modification of Bielschowsky results Nerve endings and neurofibrils: Black Neurofibrillary tangles and peripheral neuritis of neuritic plaques: Black
Seiver-Munger modification of Bielschowsky Useful for demonstrating granules of some carcinoid tumor cells
Seiver-Munger modification of Bielschowsky Argyrophil
Thioflavin S Demonstration of neurofibrillary tangles and senile plaques
Thioflavin S 10-20% NBF
Thioflavin S control CNS containing senile plaques and neurofibrillary tangles
Thioflavin S 1.Potassium permanganate 2. rinse 3. potassium meta bisulfite-oxalic acid 4.wash 5. sodium hydroxide-peroxide 6.wash 7. acetic acid 8.wash 9. 50% alcohol 10.Thioflavin S 11.50% alcohol 12. Dehydrate and clear
Thioflavin S results Alzheimer neurofibrillary tangles, senile plaques neurites, neuropil threads, senile plaque amyloid, and cerebrovascular amyloid: Bright green Diffuse plaques and extracellular tangles: Paler yellow green PSP tangles and Pick bodies: Not well demon
Mallory phosphotungstic acid hematoxylin (PTAH) Demonstration of glial cells
Mallory phosphotungstic acid hematoxylin (PTAH) 10% NBF
Mallory phosphotungstic acid hematoxylin (PTAH) 6-8 micrometers
Mallory phosphotungstic acid hematoxylin (PTAH) control Cerebral cortex
Mallory phosphotungstic acid hematoxylin (PTAH) 1. Mordant with Zenker 2. wash 3. Lugol iodine 4. decolorize 95% alcohol 5. rinse 6. Potassium permanganate 7. wash 8. Decolorize in 5% oxalic acid 9. wash 10. Stain in PTAH 11. Dehydrate and clear
Mallory phosphotungstic acid hematoxylin (PTAH) results Glial fibers: Blue Nuclei: Blue Neurons: Salmon Myelin: BLue
Holzer method Demonstration of glial cells in areas of gliosis
Holzer method Resistant to decolorization with the alkaline aniline chloroform mix
Holzer method 10% NBF
Holzer method 6-8 micrometers
Holzer method control Cerebral cortex
Holzer method 1. Phosphomolybdic acid alcohol 2. Cover sections with absolute alcohol chloroform 3. Crystal violet 4. 10% potassium bromide 5.Blot 6. Differentiate 7. Dehydrate and clear
Holzer method results Glial fibers: Blue Background: Very pale blue to colorless
Cajal stain Demonstration of astrocytes
Cajal stain Formalin ammonium bromide for no <2 days and no more than 25
Cajal stain 20-30 micrometers
Cajal stain control cerebral cortex
Cajal stain results Astrocytes with perivascular feet: Black
Cajal stain 1. Gold sublimate 2. wash 3. 5% sodium thiosulfate 4. wash 5. mount, dehydrate and clear
Weil method Demonstration of myelin sheath
Weil method Helps in diagnosing syphilis and amyotrophic lateral sclerosis
Weil method 10% NBF
Weil method 10-15 micrometers
Weil method control spinal cord or medullar
Weil method 1. staining solution 2. wash 3. 4% ferric ammonium sulfate 4. Sodium borate potassium ferricyanide 5. Wash 6. Ammonia water 7. Wash 8. Dehydrate 9. clear
Weil method results Myelin sheath: Blue to blue black Background: Light tan
Luxol Fast Blue Demonstrates myelin sheath
Luxol Fast Blue Is of the sulfonated copper phthalocyanine type, but is alcohol soluble
Luxol Fast Blue 10% NBF
Luxol Fast Blue 10-15 micrometers
Luxol Fast Blue control Spinal cord or medulla
Luxol Fast Blue 1. Luxol fast blue 2. rinse with 95% 3. rinse 4. Lithium carbonate 5. 70% alcohol 6. wash 7. Lithium to 70% several times 8. rinse 9. dehydrate and clear
Luxol Fast Blue results Myelin: Blue to blue-green Background: Colorless
Created by: Ziek98