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Histology Spring 2020 > Chapter 9: Nerve > Flashcards

Chapter 9: Nerve Flashcards

1
Q

Cresyl Echt Violet 1 Purpose

A

Identification of neurons or demonstration of lost Nissl substance that indicates neuronal injury

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2
Q

Cresyl Echt Violet 1 Principle

A

Nissl substance is very basophilic due to RNA content, and therefore stains sharply with basic aniline dyes
Adjusting pH can show both Nissl and nuclei or only Nissl

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3
Q

Cresyl Echt Violet 1 Preferred Fixative

A

10% NBF

Spinal cord control

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4
Q

Cresyl Echt Violet 1 Basic Procedure

A
  1. Deparaffinize, hydrate
  2. Stain in cresyl echt violet (ripen for 24-48 hours and filter before use)
  3. Rinse in distilled water
  4. Place sections in 95% alcohol
  5. Transfer sections to absolute alcohol
  6. Place in xylene
  7. Place in balsam-xylene mixture
  8. Differentiate in absolute alcohol, check sections microscopically
  9. Several changes of xylene
  10. repeat 7-9 multiple times until differentiation is complete; background should be colorless and blue to purple nuclei and Nissl
  11. Coverslip
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5
Q

Cresyl Echt Violet 1 Results

A

Nissl substance and nuclei: blue to purple

background: colorless

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6
Q

Cresyl Echt Violet 1 Technical notes

A

Differentiation should be repeated until the background is colorless. This usually requires several repetitions

The alcohol following the balsam-xylene mixture will become cloudy and should be changed frequently

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7
Q

Cresyl Echt Violet 2 Purpose

A

Identify neurons or loss of Nissl substance (chromatolysis)

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8
Q

Cresyl Echt Violet 2 Principle

A

Same as method one, but the cresyl echt violet is at an acidic pH which enhances contrast between the background and Nissl or nuclei

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9
Q

Cresyl Echt Violet 2 Preferred Fixative

A

10% NBF

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10
Q

Cresyl Echt Violet 2 Basic Procedure

A
  1. Deparaffinize, hydrate
  2. stain in acidic cresyl echt violet (includes alcohol and acetic acid)
  3. Dehydrate, clear, coverslip
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11
Q

Cresyl Echt Violet 2 Results

A

Nissl substance and nuclei: blue-purple

Background: colorless

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12
Q

Cresyl Echt Violet 2 Technical notes

A

Macroscopically the slides will appear unstained

Cresyl echt violet from Luxol fast blue stain may also be used to identify Nissl substance

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13
Q

Bodian Method Purpose

A

Stain nerve fibers

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14
Q

Bodian Method Principle

A
  1. Protargol (brand name silver proteinate) impregnates tissue
  2. Copper is added to impregnating solution to “destain” connective tissue to improve differentiation
  3. Hydroquinone reduces the deposited silver salts into visible metallic silver
  4. Gold chloride is the toner (oxalic acid can be used to reduce the gold and intensify the stain)
  5. Sodium thiosulfate removes unreduced silver
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15
Q

Bodian Method Preferred Fixative

A

10% NBF

Control: peripheral nerve or cerebral cortex to avoid cross-section

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16
Q

Bodian Method Basic Procedure

A
  1. Deparaffinize, hydrate
  2. Add clean copper shot to Protargol at 37C for 48 hours
  3. Rinse in distilled water
  4. place in reducing solution
  5. Rinse in water
  6. Tone in gold chloride
  7. rinse in water
  8. Develop in Oxalic Acid, checking microscopically until the background is grey and the nerve fibers appear clearly stained. Don’t treat too long or it will ruin the silver
  9. Rinse in water
  10. Treat with sodium thiosulfate to remove unreduced silver
  11. Rinse in distilled water
  12. Counterstain, if desired, with aniline blue
  13. Dehydrate, clear, coverslip
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17
Q

Bodian Method Results

A

Nerve fibers and nuclei: black

Background: light grey or blue (if counterstained)

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18
Q

Bodian Method Technical notes

A

Copper shot is cleaned with aqua regia (HCl +Nitric Acid). After use it should be gradually poured into a very large volume of water and then discarded in the sink. Do not pour directly into sink, do not add water to the acid.

Protargol should be left undisturbed until it is completely dissolved

use chemically clean glassware and non-metallic forceps
Don’t overcounterstain with Aniline blue

You can also use nuclear fast red as a counterstain

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19
Q

Holmes Silver Nitrate Method Purpose

A

Demonstrate nerve fibers and neurofibrils

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20
Q

Holmes Silver Nitrate Method Principle

A

Bodian doesn’t work well because the Protargol solution never reaches necessary alkalinity for proper impregnation. Holmes developed a buffered impregnation solution

This is an argyrophil silver method that requires chemical reduction
Gold chloride toner, Oxalic acid reducer, and sodium thiosulfate are the same as in Bodian

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21
Q

Holmes Silver Nitrate Method Preferred Fixative

A

10% NBF
Control: peripheral nerve or cerebral cortex to avoid cross-section
10-15uM paraffin

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22
Q

Holmes Silver Nitrate Method Basic Procedure

A
  1. Deparaffinize, hydrate
  2. Place in 20% silver nitrate in the dark
  3. Prepare impregnating solution (Boric acid, Borax, water, silver nitrate, pyridine)
  4. Wash slides in distilled water
  5. Place slides in impregnating solution overnight at 37C
  6. Remove slides, shake of excess fluid, and place in reducer (hydroquinone, sodium sulfite, water)
  7. wash in running water
  8. Rinse in distilled water
  9. Tone in gold chloride (can be re-used until brown precipitate forms or the solution becomes cloudy)
  10. Rinse in distilled water
  11. Place slides in oxalic acid, when the axons are thoroughly blue-black, stop the process
  12. Rinse in distilled water
  13. place in sodium thiosulfate
  14. wash in tap water. Counterstain if desired
  15. Dehydrate, clear, coverslip
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23
Q

Holmes Silver Nitrate Method Results

A

Axons and nerve fibers: black

Neurofibrils: black

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24
Q

Holmes Silver Nitrate Method Technical notes

A

Pyridine is toxic by ingestion, inhalation, and skin absorption. Use under a fume hood with gloves and goggles

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25
Q

Bielschowsky-PAS Purpose

A

To demonstrate nerve fibers, neurofibrillary tangles, and senile plaques in Alzheimer disease

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26
Q

Bielschowsky-PAS Principle

A
  1. impregnate with ammoniacal silver which deposits on neurofibrils and axons
  2. silver is reduced to metallic silver by formaldehyde in the developer
  3. Gold chloride tones the tissue and removes yellow
  4. sodium thiosulfate removes unreduced silver background
  5. schiff reaction (PAS) stains basement membranes and amyloid in the plaques
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27
Q

Bielschowsky-PAS Preferred Fixative

A

10% NBF

Control: CNS tissue containing plaques and tangles

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28
Q

Bielschowsky-PAS Basic Procedure

A
  1. Prepare ammoniacal silver solution
  2. Deparaffinize, hydrate
  3. Place in silver nitrate solution in the dark at room temp
  4. Wash with distilled water
  5. Place in ammoniacal silver solution
  6. Wash in ammonia water
  7. While slides are in ammonia water, add 2 drops of developer to the ammoniacal silver solution from step 5
  8. place slides in ammoniacal/developer solution. The tissue should turn brown in about 3 minutes
  9. Wash well in ammonia water, then distilled water
  10. Tone in gold chloride until grey appears
  11. Wash in ammonia water, then rinse in distilled water
  12. Place in sodium thiosulfate
  13. Wash in running tap water
  14. Rinse well in distilled water
  15. Place sections in 1% periodic acid
  16. Rinse in distilled water
  17. Place in schiff reagent
  18. Wash in tap water
  19. Dehydrate, clear, coverslip
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29
Q

Bielschowsky-PAS Results

A

Neurofibrillary tangles: dark black
Peripheral neurites and plaques: dark black
Axons: black
Amyloid (plaque cores and vasculature): magenta
Lipofuchsin: magenta

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30
Q

Bielschowsky-PAS Microwave Purpose

A

Demonstrate nerve fibers, tangles, and plaques in Alzheimer’s

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31
Q

Bielschowsky-PAS Microwave Principle

A

Sections are not toned with gold chloride so yellow background remains

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32
Q

Bielschowsky-PAS Microwave Preferred Fixative

A

10% NBF

Control: CNS with plaques and tangles

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33
Q

Bielschowsky-PAS Microwave Basic Procedure

A
  1. Deparaffinize, hydrate
  2. Place slides in 1% Silver Nitrate, dip several times and incubate in warm solution for 15 minutes
  3. Place slides in distilled water
  4. 1% silver nitrate from step 2 + 28% ammonium drops until precipitate clears, then 5% silver nitrate drops until solution becomes slightly cloudy
  5. microwave slides in the solution, dip slides and incubate in warm solution
  6. Place slides in 1% ammonium hydroxide
  7. add 3 drops developer to ammoniacal silver from step 5, place slides in solution until tissue turns brown
  8. Place slides in 1% ammonium hydroxide
  9. Rinse in distilled water
  10. Wipe silver mirror off both sides of slide without disturbing the tissue
  11. place slides in 2% sodium thiosulfate
  12. Rinse slides in distilled water
  13. Dehydrate, clear, coverslip
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34
Q

Bielschowsky-PAS Microwave Results

A 
Axons: brown to black
Cytoplasmic neurofibrils: brown to black
Tangles and plaques: dark brown or black
Neuromelanin: black
Lipofuchsin: brown or black
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35
Q

Bielschowsky-PAS Microwave Technical notes

A

Use chemically cleaned glassware, non-metallic forceps

Uses much less silver nitrate than traditional Bielschowsky and stains tangles and plaques better

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36
Q

Sevier-Munger Modification of Bielschowsky Purpose

A

Demonstrate nerve fibers, tangles, and plaques for Alzheimer’s

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37
Q

Sevier-Munger Principle

A

Ammoniacal silver impregnation
Reduced by formaldehyde developer
No toning with gold chloride so yellow background remains
Sodium Thiosulfate removes unreduced silver

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38
Q

Sevier-Munger Preferred Fixative

A

10% NBF
Control: CNS with tangles and plaques
6-8uM sections

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39
Q

Sevier-Munger Basic Procedure

A
  1. Deparaffinize, hydrate
  2. Preheat 20% silver nitrate, then add slides to warm silver solution and incubate in the oven
  3. Rinse one slide at a time in distilled water and place in a clean, dry staining jar
  4. add 10 drops formalin, while agitating, to working ammoniacal silver solution, pour over slides and develop until golden brown. Check microscopically, do not wash while checking, keep in motion while developing to avoid precipitation
  5. Rinse slides well in tap water
  6. Place in sodium thiosulfate
  7. Wash well in tap water
  8. Dehydrate, clear, coverslip
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40
Q

Sevier-Munger Results

A

Nerve endings and neurofibrils: black

Tangles, peripheral neurites, and plaques: black

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41
Q

Sevier-Munger Technical notes

A

Very reliable and reproduce-able technique

Concentration of ammonium hydroxide and formalin are critical
leave a few particles of silver, do not add extra ammonia

This is an argyrophil stain that is also useful fro demonstrating the granules of some carcinoid tumor cells

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42
Q

Thioflavin S (modified) Purpose

A

Demonstrates presence of tangles, plaques, threads in addition to parenchymal and amyloid deposition in Alzheimer’s disease

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43
Q

Thioflavin S (modified) Principle

A

Fluorescent visualization of amyloid
Pretreatment with potassium permanganate, and bleaching with potassium metabisulfate and oxalic acid
Treatment with sodium hydroxide and hydrogen peroxide, which remove lipid autofluorescence for better definition of pathological lesions
Visualization is better than routine Thioflavin S ans is not affected by prolonged fixation

More sensitive than silver methods
Faster and cheaper than silver

44
Q

Thioflavin S (modified) Preferred Fixative

A

10-20% NBF
Control: CNS with plaques and tangles
6um sections air dried overnight, then 10 minutes in 60C oven

45
Q

Thioflavin S (modified) Basic Procedure

A
  1. Deparaffinize, hydrate
  2. Rinse and old in distilled water
  3. cover tissue with 0.25% potassium permanganate
  4. wash slides in running tap water
  5. treat slides with 1% potassium bisulfate
  6. wash in running tap water
  7. place in sodium hydroxide-peroxide
  8. wash in running tap water and then filtered water
  9. place slides in 0.25% acetic acid
  10. wash slides in running tap water
  11. place slides in 50% alcohol
  12. place in Thioflavin S
  13. rinse slides in 50% alcohol with agitation
  14. Rinse in 95% alcohol
  15. completely dehydrate in absolute alcohol and clear in xylene. Mount with non-fluorescent mounting medium
  16. View slides on a fluorescent filter set that incorporates a blue-violet excitation filter
46
Q

Thioflavin S (modified) Results

A

tangles, neurites, threads, plaque amyloid, and cerebrovascular amyloid: bright green

Diffuse plaques and extracellular plaques: paler yellow green

PSP tangles and Pick bodies: not well demonstrated

47
Q

Thioflavin S (modified) Technical notes

A

Float tissue on pre-heated DI water bath

Mount sections with cytoseal 60

Staining is stable for several months at room temp

48
Q

PTAH Purpose

A

Demonstration of glial fibers (also used to demonstrate muscle striations)

49
Q

PTAH Principle

A

High ratio of PTA to hematein 20:1 so that tungsten binds all available hematein to form a blue lake which stains glial fibers, nuclei, and myelin. Red brown or salmon color of neurons is due to the PTA alone. Overuse of alcohol will fade the red-brown

50
Q

PTAH Preferred Fixative

A

10% NBF
6-8uM sections
Control: Cerebral cortex (not spinal cord)

51
Q

PTAH Basic Procedure

A
  1. Deparaffinize, hydrate
  2. Mordant the sections overnight at room temp in Zenker solution containing acetic acid
  3. Wash sections in running water
  4. Place in Lugol Iodine. Do not use sodium thiosulfate because it may impair subsequent staining
  5. Decolorize the sections in 95% alcohol for 1 hour
  6. Rinse rapidly in distilled water
  7. Place in 1% potassium permanganate
  8. Wash in running tap water
  9. Decolorize the sections in 5% oxalic acid
  10. Wash in running tap water
  11. Stain in PTAH overnight
  12. Dehydrate, clear, coverslip
52
Q

PTAH Results

A

Glial fibers: blue
Nuclei: blue
Neurons: salmon
Myelin: blue

53
Q

PTAH Technical notes

A

Replaced by immunohistochemical methods

Don’t over rinse in EtOH

Don’t use sodium thiosulfate or stain won’t work

54
Q

Holzer Method Purpose

A

To demonstrate glial fibers and areas of gliosis (glial damage)

55
Q

Holzer Method Principle

A

Glial fibers are stained with crystal violet and are resistant to decolorization with an alkaline aniline-chloroform mixture

56
Q

Holzer Method Preferred Fixative

A

10% NBF
6-8uM sections
Control: Cerebral cortex, not spinal cord

57
Q

Holzer Method Basic Procedure

A
  1. Deparaffinize, hydrate
  2. place in PMA-alcohol
  3. Drain off excess fluid, place slides in a staining rack and cover the sections with absolute alcohol-chloroform mixture. The tissue should become translucent
  4. While sections are still wet, cover them with the crystal violet stain
  5. Replace the stain with 10% potassium bromide
  6. Blot the sections dry and then allow them to air dry
  7. Differentiate in aniline-oil+chloroform+ammonium hydroxide differentiating solution
  8. Wash in several changes of xylene. Repeat steps 7 and 8 until the background is very pale or colorless
  9. Mount with synthetic resin
58
Q

Holzer Method Results

A

Glial fibers: blue

Background: very pale blue to colorless

59
Q

Holzer Method Technical notes

A

Crystal Violet precipitate may be removed with straight aniline oil

Aniline oil and chloroform are both very hazardous; use in a chemical fume hood

The aniline oil-chloroform-ammonium hydroxide differentiating solution resists decolorization

60
Q

Cajal Method Purpose

A

Demonstrate astrocytes

61
Q

Cajal Method Principle

A

Astrocytes are selectively stained with the Cajal gold sublimate method on frozen sections

62
Q

Cajal Method Preferred Fixative

A

Formalin ammonium bromide for 2-25 days, rinse and place in formalin ammonium bromide for 2 days if previously fixed with 10% NBF

20-30uM Sections, free floating, no slides
Control: cerebral cortex, not spinal cord

63
Q

Cajal Method Basic Procedure

A
  1. wash free floating frozen sections in several changes of distilled water
  2. Transfer the sections to the gold sublimate solution (gold chloride + mercuric chloride) and incubate in the dark for 4 hours, sections should turn purple
  3. Wash well in distilled water
  4. Treat with 5% sodium thiosulfate
  5. Wash in distilled water
  6. Carefully mount the sections on slides, blot and dehydrate
  7. Clear in xylene and mount with synthetic resin
64
Q

Cajal Method Results

A

Astrocytes: black

65
Q

Cajal Method Technical notes

A

Tissue sections better if washed in tap water for 30 minutes before freezing

chemicals must be pure; brown chloride is preferred over yellow gold chloride

Protoplasmic (grey matter) astrocytes lose stainability after prolonged fixation

Staining solution should not exceed 30C

mixture of mercuric chloride and gold chloride is essential for astrocyte demonstration

mercuric chloride is extremely toxic and an environmental hazard

66
Q

Weil’s Purpose

A

Demonstration of myelin

Myelin degeneration, such as in syphilis

67
Q

Weil’s Principle

A

Mordant-hematoxylin solutions attaches to the phospholipid component of the myelin sheath, which has an affinity for the cationic (+) lake
Regressive technique
1. Ferric ammonium sulfate: excess mordant differentiation removes most of the excess dye
2. Borax ferricyanide: oxidizer differentiation which removes any remaining non-specifically bound hematoxylin lake and forms a colorless oxidation product.

Only myelin sheath and red blood cells remain stained

68
Q

Weil’s Preferred Fixative

A

10% NBF
10-15uM Sections
Control: Spinal cord or medulla

69
Q

Weil’s Basic Procedure

A
  1. Deparaffinize, hydrate
  2. Transfer sections to staining solution for 30 minutes at 54-56C
  3. Wash in tap water
  4. Differentiate in 4% ferric ammonium sulfate until the gray matter can just be distinguished from the white matter and the stain is removed from the slides
  5. Wash in tap water
  6. Complete differentiation in the sodium borate-potassium ferricyanide solution. This should be checked until the grey and white matter are sharply defined
  7. Wash in tap water
  8. Treat sections in diluted ammonia water
  9. Wash in distilled water
  10. Dehydrate, clear, coverslip
70
Q

Weil’s Results

A

Myelin sheath: blue to blue-black

Background: light tan

71
Q

Weil’s Technical notes

A

Gray matter and demyelinated white matter should appear light brown and contrast sharply with blue-black myelinated white matter

Easy macroscopic differentiation

Naturally ripen for 6 months or chemically ripen with ferric ammonium sulfate

72
Q

Luxol Fast Blue Purpose

A

Demonstrate myelin

73
Q

Luxol Fast Blue Principle

A

Similar to alcian blue, but is alcohol rather than water soluble. Staining is accomplished through an acid-base reaction in which the base of the myelin lipoprotein swaps with the base of the dye

74
Q

Luxol Fast Blue Preferred Fixative

A

10% NBF
10-15uM Sections
Control: spinal cord or medulla

75
Q

Luxol Fast Blue Basic Procedure

A
  1. Deparaffinize, hydrate
  2. Place slides in LFB (Luxol Fast Blue) and incubate overnight at 56-58C
  3. Rinse in 95% alcohol to remove excess stain (LFB is alcohol soluble)
  4. Rinse in water
  5. Begin differentiating by immersing slides in lithium carbonate solution
  6. Continue differentiation in 70% alcohol until gray matter and white matter can be distinguished. Do not over differentiate
  7. Wash in distilled water
  8. Finish differentiation by rinsing briefly in lithium carbonate and then through several changes of 70% alcohol until the greenish blue of the white matter contrasts sharply with colorless grey matter
  9. Rinse in distilled water
  10. Dehydrate, clear, coverslip
76
Q

Luxol Fast Blue Results

A

Myelin: blue to blue-green
Background: colorless

77
Q

Luxol Fast Blue Technical notes

A

Gray matter and demylinated white matter should be almost colorless and contrast sharply with blue stained myelinated white matter

Quality of the stain (contrast) can be determined macroscopically

Differentiate until gray matter is almost colorless

78
Q

LFB-Holmes silver nitrate Purpose

A

Demonstrate myellin and nerve fibers in the same section

79
Q

LFB-Holmes silver nitrate Principle

A

LFB staining is accomplished through an acid-base reaction in which the base of the myelin lipoprotein swaps with the base of the dye

Holmes is an argyrophil silver method that requires chemical reduction
Gold chloride toner, Oxalic acid reducer, and sodium thiosulfate

80
Q

LFB-Holmes silver nitrate Preferred Fixative

A

10% NBF
10-15uM Sections
Control: Cerebral cortex or longitudinal section of peripheral nerve

81
Q

LFB-Holmes silver nitrate Basic Procedure

A
  1. Deparaffinize, hydrate
  2. 20% silver nitrate in the dark at room temp
  3. make impregnating solution (boric acid+borax+silver nitrate+pyradine)
  4. Take slides out of 20% silver nitrate and wash in distilled water
  5. 37C overnight incubation in impregnating solution
  6. shake off excess fluid, and place in reducer (hydroquinone+sodium sulfite)
  7. Wash in running water, then distilled water
  8. Tone in gold chloride. Solution may be reused until a brown precipitate forms or the solution becomes cloudy
  9. Rinse in distilled water
  10. Place in oxalic acid, stop when sections are blue-black
  11. Rinse in distilled water
  12. Place slides in sodium thiosulfate
  13. Wash in tap water
  14. place in 95% alcohol
  15. Stain in LFB overnight at 60C
  16. place in 95% alcohol
  17. Place in distilled water
  18. Place in lithium carbonate
  19. Differentiate in 70% alcohol
  20. Rinse in distilled water (repeat steps 18-20 until desired differentiation is achieved)
  21. Dehydrate, clear, coverslip
82
Q

LFB-Holmes silver nitrate Results

A

Myelin: blue to green

Axons and nerve fibers: black

83
Q

LFB-cresyl echt violet Purpose

A

Demonstrate both myelin and Nissl substance

84
Q

LFB-cresyl echt violet Principle

A

LFB staining is accomplished through an acid-base reaction in which the base of the myelin lipoprotein swaps with the base of the dye

Cresyl echt violet uses a basic aniline dye to stain RNA blue, thus demonstrating the RNA in the rough endoplasmic reticulum which comprises the Nissl substance

85
Q

LFB-cresyl echt violet Preferred Fixative

A

10% NBF
10-15uM sections
Control: spinal cord or medulla

86
Q

LFB-cresyl echt violet Basic Procedure

A
  1. Deparaffinize, hydrate
  2. Place slides in LFB, incubate overnight in 56-58C
  3. rinse in 95% alcohol to remove excess stain
  4. Rinse in distilled water
  5. Begin differentiation by immersion in lithium carbonate
  6. Continue differentiation in 70% alcohol until grey and white matter can be distinguished
  7. Rinse in distilled water
  8. Finish differentiation by rinsing briefly in lithium carbonate and then through several changes of 70% alcohol until sharp contrast is achieved
  9. Rinse in distilled water
  10. Add acetic acid, filter, and preheat cresyl echt violet. Place slides in cresyl echt violet for 6 minutes, keep warm while staining.
  11. Differentiate in several changes of 95% alcohol
  12. Dehydrate, clear, coverslip
87
Q

LFB-cresyl echt violet Results

A

Myelin: blue (from LFB)
Nissl substance: violet (from cresyl)
Nuclei: violet (from cresyl)

88
Q

LFB-cresyl echt violet Technical notes

A

If you don’t add acetic acid to the CEV, you will get a diffuse violet background stain

Staining of Nissl will be reduced if you don’t heat the CEV prior to staining

CEV counterstain intensifies the myelin sheath stain

89
Q

LFB-PAS-Hematoxylin Purpose

A

Demonstrate myelin sheath, basement membranes, senile plaques, fungi, and corpora amylacea
The various stains sharpen and complement each other

90
Q

LFB-PAS-Hematoxylin Principle

A

LFB staining is accomplished through an acid-base reaction in which the base of the myelin lipoprotein swaps with the base of the dye

Periodic acid oxidizes reactive groups to form aldehydes (-CHO)
Basic fuschin + sulfurous acid = leucofuschin/Schiff’s reagent which binds to exposed aldehyde groups
Water washes away the sulfur resulting in the rose chromophore

91
Q

LFB-PAS-Hematoxylin Preferred Fixative

A

10% NBF
10-15uM Sections
Control: Cerebral cortex or medulla

92
Q

LFB-PAS-Hematoxylin Basic Procedure

A
  1. Deparaffinize, hydrate
  2. Incubate in LFB overnight at 56-58C
  3. Rinse in 95% alcohol to remove excess stain
  4. Rinse in distilled water
  5. Begin differentiation by placing in lithium carbonate
  6. Continue differentiation in 70% alcohol until grey and white matter become distinguished
  7. Wash in distilled water
  8. Finish differentiation by rinsing briefly in lithium carbonate and then through several changes of 70% alcohol until sharp contrast is achieved (repeat 7-8 as necessary)
  9. Rinse in distilled water
  10. Place in periodic acid (aldehyde formation)
  11. Place in distilled water
  12. Place in schiff solution
  13. Wash in tap water
  14. Stain with Harris hematoxylin
  15. Wash in tap water (if background is not clear dip once in acid-alcohol and wash, if nuclei are not dark blue to purple dip briefly in dilute ammonium hydroxide and wash)
  16. Dehydrate, clear, coverslip
93
Q

LFB-PAS-Hematoxylin Results

A 
Capillary basement membranes: Rose
Fungi: Rose
Corpora amylacea: Rose
Senile plaques: Rose
Myelin Sheath: Blue to blue-green
Nuclei: purple
94
Q

Two anatomic parts of the nervous system

A

CNS: brain and spinal cord
PNS: everything else

95
Q

2 functional parts of the nervous system

A

Somatic: voluntary
Autonomic: involuntary

96
Q

3 groups of nervous system stains based on stain target

A

Neuronal cell bodies and processes
Glial cells and processes
Myelin sheath

97
Q

Neuron

A

Cellular unit of the nervous system, conduct information vial electrochemical impulses

98
Q

What are the structural components of a neuron?

A 
Cell body containing nucleus
Dendrites
Axon
Myelin sheath
Node of Ranvier
99
Q

Characteristics of Nissl Substance

A

Basophilic: stains blue/black with hematoxylin, or basic aniline dyes like thionin and cresyl echt violet
Large aggregates of rough endoplasmic reticulum with RNA content being stained

100
Q

Neuroglia

A

“Nerve glue” like a connective tissue for the CNS
Insulate neurons except at synapses
produce the myelin sheath

101
Q

4 types of glial cells

A

Oligodendroglia/oligodendrocytes
Astrocytes
Microglia
Ependymal Cells

102
Q

Oligodendroglia

A

Produce and maintain myelin sheath in the CNS

103
Q

Astrocytes

A

Protoplasmic(gray) and fibrous(white)
help in scar formation after brain injury
support nerve fibers
Star shaped (stellate)

104
Q

Microglia

A

fixed phagocytic cells found throughout the brain and spinal cord (immune response)

105
Q

Ependymal Cells

A

Epithelium with a brush border that acts as the blood/brain barrier

106
Q

Myelin

A

Fatty insulation around neurons
Stains with lipid stains
Deposited by oligodendrocytes on multiple neurons in the CNS
Deposited by Schwann cells on individual neurons in the PNS
Typically demonstrated with Luxol fast blue and iron hematoxylin

Histology Spring 2020 (14 decks)

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