Pigments & Minerals Staining

Front 1

Prussian blue purpose

Back 1

Detect ferric iron (Fe^3+)

Front 2

Prussian blue reagents

Back 2

Potassium ferrocyanide

Hydrochloric acid

Nuclear fast red - counterstain

Front 3

Prussian blue results

Back 3

Ferric iron - blue

Nuclei, hemofuchsin - bright red

Background - pink

Front 4

Prussian blue control

Back 4

Tissue containing iron (small amounts)

Front 5

Prussian blue fixative

Back 5

Alcohol or 10% NBF

Front 6

Prussian blue cut

Back 6

4-5 microns

Back 7

Prussian blue

Front 8

Turnbull blue purpose

Back 8

Demonstrate ferrous iron (fe^2+)

Front 9

Turnbull blue reagents

Back 9

Potassium ferricyanide

HCl

Acetic acid - wash

Nuclear fast red - counterstain

Front 10

Turnbull blue results

Back 10

Ferrous iron - blue

Background - pink/red

Front 11

Turnbull blue control

Back 11

Tissue containing ferrous iron

Front 12

Turnbull blue fixative

Back 12

Alcohol or 10% NBF

Front 13

Turnbull blue cut

Back 13

4-5 microns

Front 14

Turnbull blue troubleshoot notes

Back 14

Use nonmetallic forceps

Chemically clean glasswear

DI water

Front 15

Front (Term)

Back 15

Turnbull blue

Front 16

Bile (halls) stain purpose

Back 16

Demonstrate bilirubin

differentiate between bile pigment from other pigment

Front 17

Bile (halls) stain reagent

Back 17

Fouchet reagent - ferric chloride (oxidizer) & trichloroacetic Acid

Vangieson solution - counterstain

Front 18

Bile (halls) stain results

Back 18

Bile, bilirubin - green

Collagen/mucin - Red

Background/muscle - yellow

Front 19

Bile (halls) stain control

Back 19

Bile

Front 20

Bile (halls) stain fixative

Back 20

10% NBF

Front 21

Bile (halls) stain cut

Back 21

4-5 microns

Front 22

Melanin is...

Back 22

Endogenous

Nonlipidic

Nonhematogenous

Front 23

Front (Term)

Back 23

Bile (halls) stain

Front 24

Schmorl stain purpose

Back 24

Indication of reducing substances

Melanin, argentaffin granules, formalin pigment

Front 25

Schmorl stain reagents

Back 25

Ferric chloride - reducing substance

Potassium ferricyanide - ferrous iron reaction to form ferrous ferricyanide

Mayer mucicarmine - stains mucin red

Metanil yellow - counterstain

Front 26

Schmorl stain results

Back 26

Reducing substances- blue/green

Goblet cells/mucin - Red

Background - yellow

Front 27

Front (Term)

Back 27

Schmorl stain

Front 28

Schmorl stain control

Back 28

Melanin or argentaffin

Front 29

Schmorl stain fixative

Back 29

10% NBF

Front 30

Schmorl stain cut

Back 30

4-5 microns

Front 31

Schmorl stain troubleshooting

Back 31

Works well on GI tract

Can use nuclear fast red as counterstain

Use clean glasswear

Use fresh sol (nonspecific staining)

Front 32

Front (Term)

Back 32

Schmorl stain

Front 33

Fontana-Masson stain purpose

Back 33

Demonstrate argentaffin substances (melanin, argentaffin granules, carcinoid tumor, neurosecretary granules)

Front 34

Fontana-Masson reagents

Back 34

Silver sol - impregnation

Gold chloride- toner

Sodium thiosulfate- remove silver

Nuclear fast red - counterstain

Front 35

Fontana-Masson results

Back 35

Melanin, argentaffin- black

Background - pink

Front 36

Fontana-Masson control

Back 36

Skin - melanin

Small intestine/appendix-argentaffin

Front 37

Fontana-Masson fixative

Back 37

10% NBF (avoid alcohol)

Front 38

Front (Term)

Back 38

Fontana-Masson

Front 39

Fontana-Masson troubleshooting

Back 39

Avoid overstaining

Clean glasswear

Non metallic forceps

Demonstrate melanin w/ bleach

Front 40

Front (Term)

Back 40

Fontana-Masson overstained

Front 41

Grimelius purpose

Back 41

Demonstrate agyrophilic granules- neurosecretory

Front 42

Grimelius reagents

Back 42

Silver solution - impregnation

Reducing sol - makes silver visible (hydroquinone & sodium sulfite)

Nuclear fast red - counterstain

Front 43

Grimelius results

Back 43

Argentaffin granules-dark brown to Black

Agyrophilic granules- dark brown to black

Nuclei - Red

Background - pale yellow/brown

Front 44

Grimelius control

Back 44

Carcinoid tumor or small intestine (agyrophilic positive)

Front 45

Grimelius fixative

Back 45

10% NBF

Front 46

Grimelius cut

Back 46

4-5 microns

Front 47

Grimelius troubleshooting

Back 47

Clean glasswear & nonmetallic forceps needed

Front 48

Front (Term)

Back 48

Grimelius

Front 49

Cherukian-Schenk purpose

Back 49

Agyrophilic granules in neurosecretory tumors

Front 50

Cherukian-Schenk reagents

Back 50

Hydrate acidified water - citric Acid

Silver nitrate - impregnation

Reducing sol - makes silver visible (hydroquinone, sodium sulfite)

Nuclear fast red - counterstain

Front 51

Cherukian-Schenk results

Back 51

Agyrophilic & argentaffin granules- Black

Nuclei - Red

Background - yellow/brown

Front 52

Cherukian-Schenk control

Back 52

Carcinoid tumor, small intestine

Front 53

Cherukian-Schenk fixative

Back 53

10% NBF

Front 54

Cherukian-Schenk cut

Back 54

4-5 microns

Front 55

Cherukian-Schenk troubleshooting

Back 55

Clean glassware non metallic forceps

Front 56

Front (Term)

Back 56

Cherukian-Schenk

Front 57

Gomori-methenamine silver (GMS) purpose

Back 57

demonstrate urates

Front 58

Gomori-methenamine silver (GMS) reagents

Back 58

Impregnated sol - silver nitrate, methenamine sol, sodium borate

Sodium thiosulfate- rinse unbound silver

Light green - counterstain

Front 59

Gomori-methenamine silver results

Back 59

Urates - Black

Background - green

(Birefringent)

Front 60

Von kossa troubleshooting

Back 60

Use chemically clean glassware

Non metallic forceps

Removal of mercury pigment = removal of calcium salts (false negative)

Front 61

Alizarin Red S purpose

Back 61

Calcium demonstration

Front 62

Alizarin Red S tips

Back 62

Birefringent

Use light green as counterstain

Front 63

Rhodanine purpose

Back 63

Demonstrate copper (Wilson’s disease)

Front 64

Rhodanine reagents

Back 64

Rhodanine - binds copper protein

Mayer’s heme - nuclear stain

Sodium borate - bluing agent

Front 65

Rhodanine reuslts

Back 65

Copper - Red

Nuclei - light blue

Front 66

Rhodanine control

Back 66

Section containing Copper (fetal liver)

Front 67

Rhodanine fixative

Back 67

10% NBF

Front 68

Rhodanine cut

Back 68

6-8 microns

Front 69

Rhodanine troubleshooting

Back 69

Avoid overstaining with hematoxylin

Front 70

Gomori-methenamine silver control

Back 70

Urates

Front 71

Gomori-methenamine silver fixative

Back 71

Absolute alcohol!!! (Crystals desolve in aqueous)

Front 72

Gomori-methenamine silver cut

Back 72

4-5 microns

Front 73

Von kossa purpose

Back 73

Demonstrate calcium

Front 74

Von kossa reagents

Back 74

Silver nitrate - reacts with calcium salts (bright lights)

Sodium thiosulfate- removes I reduced silver

Nuclear fast red - counterstain

Front 75

Von kossa results

Back 75

Calcium salts - Black

Background - Red

Front 76

Von kossa control

Back 76

Calcium

Front 77

Von kossa fixative

Back 77

Alcohol preferred, 10% NBF okay

Front 78

Von kossa cut

Back 78

4-5 microns

Front 79

Alizarin Red S purpose

Back 79

Demonstrate calcium

Front 80

Alizarin Red S fixative

Back 80

Alcoholic formalin or 10% NBF

Front 81

Alizarin Red S control

Back 81

Calcium

Front 82

Alizarin Red s cut

Back 82

4-5 microns

Front 83

Rhodanine purpose

Back 83

Detection of copper (Wilson’s disease)

Front 84

Rhodanine fixative

Back 84

10% NBF

Front 85

Rhodanine cut

Back 85

6-8 microns

Front 86

Rhodanine control

Back 86

Copper

Front 87

Rhodanine results

Back 87

Copper - bright red to red yellow

Nuclei - light blue