Pigment And Mineral Stains

Front 1

Name three exogenous pigments.

Back 1

Carbon/anthracotic
Asbestos fibers
Tattoo pigments

Front 2

Prussian blue/ferric stain protocol

Back 2

D+H to DW
2% potassium ferrocyanide-HCl solution
DWW
NFR
RWW
DCM

Front 3

Results of the Prussian/Fe stain

Back 3

Nuclei & hemofuchsin: bright red
Hemosiderin: blue
Background: pink

Front 4

Potassium ferrocyanide, 2% solution

Back 4

Potassium ferrocyanide, 10 g
DW, 500 mL

Front 5

Hydrochloric acid solution, 2%

Back 5

Concentrated HCl, 10 mL
DW, 490 mL

Front 6

NFR solution (Kernechtrot)

Back 6

NFR, 0.5 g
Aluminum sulfate, 25 g
DW, 500 mL

Front 7

Turnbull blue stain for ferrous iron protocol

Back 7

Depar + hydrate to DW
Ferricyanide staining solution, 1 hour
1% acetic acid
NFR, 5 min
DWW
DCM

Front 8

Results for Turnbull blue stain

Back 8

Ferrous iron: blue
Background: pink-red

Front 9

Hydrochloric acid, 0.06N solution

Back 9

Concentrated HCl, 2.5 mL
DW, 497.5 mL

Front 10

Potassium ferricyanide staining solution

Back 10

Potassium ferricyanide, 0.4 g
HCl, 0.06N solution, 40 mL
Prepare fresh just before use

Front 11

Acetic acid, 1% solution

Back 11

GAA, 1 mL
DW, 100 mL

Front 12

Schmorl technique for reducing substances protocol

Back 12

Depar and hydrate to DW
Ferric chloride-potassium ferricyanide working solution, 3 changes
DWW
Working Mayer mucicarmine
DWW
Metanil yellow
DWW
DCM

Front 13

Results for Schmorl technique

Back 13

Reducing substances: blue-green
Goblet cells, mucin: rose
Background: yellow-green

Front 14

Silver nitrate 10% solution

Back 14

Silver nitrate, 10 g
DW, 100 mL

Front 15

Fontana silver solution

Back 15

25 mL 10% silver nitrate
Swirl, add concentrated ammonium hydroxide drops
Add until solution clouds, then clears
Add 10% silver nitrate dropwise until permanent turbidity
Filter and dilute 20 mL solution:40 mL DW

Front 16

Gold chloride, 0.2% solution

Back 16

Gold chloride, 1% solution 10 mL
DW, 40 mL

Front 17

Ferric chloride, 1% stock solution

Back 17

Ferric chloride, 1.5 g
DW, 150 mL

Front 18

Potassium ferricyanide stock solution, 0.1%

Back 18

Potassium ferricyanide, 0.1g
DW, 100 mL

Front 19

Ferric chloride-potassium ferricyanide working solution

Back 19

Ferric chloride stock solution, 150 mL
Potassium ferricyanide stock solution, 50 mL
Adjust pH to 2.4 with 1N HCl

Front 20

Metanil yellow, 0.25% solution

Back 20

Metanil yellow, 0.25 g
DW, 100 mL
GAA, 0.25 mL

Front 21

Fontana-Masson stain for melanin and argentaffin granules protocol

Back 21

Depar and hydrate to DW
Silver nitrate solution, 56 deg C
DWW
0.2% gold chloride solution
DWW
5% hypo
DWW
NFR
DWW
DCM

Front 22

Fontana-Masson results

Back 22

Melanin: black
Argentaffin granules: black
Nuclei: pink

Front 23

Grimelius argyrophil stain protocol

Back 23

Depar and hydrate to DW
Room temp working silver solution, incubate @60 deg C
DWW
Reducing solution, 40-50 deg C
DWW
Working silver solution, 60 deg C
DWW
Reducing solution
DWW
NFR
DWW
DCM

Front 24

Grimelius results

Back 24

Argentaffin granules, argyrophil granules: dark brown to black
Nuclei: red
Background: pale yellow-brown

Front 25

Silver nitrate, 1% solution

Back 25

Silver nitrate, 0.1g
DW, 10 mL
Prepare fresh each time

Front 26

Acetic acid, 0.2M solution

Back 26

GAA, 6 mL
DW, 94 mL

Front 27

Sodium acetate, 0.2M solution

Back 27

Sodium acetate, trihydrate 2.75 g
DW, 100 mL
Store in fridge

Front 28

Acetic acid-Sodium acetate buffer, 0.2M solution

Back 28

Acetic acid, 0.2M solution 5 drops
Sodium acetate, 0.2M solution, 18 mL
Adjust pH to 5.6, make fresh each time

Front 29

Working silver solution

Back 29

Acetic acid-acetate buffer, pH 5.6, 5 mL
DW, 48.5 mL
SIlver nitrate, 1% solution
Prepare fresh each time

Front 30

Reducing solution, Grimelius

Back 30

Hydroquinone, 0.5 g
Sodium sulfite, anhydrous, 1.25 g
DW, 50 mL
Prepare just before use

Front 31

Churukian-Schenk method for argyrophil granules protocol

Back 31

Depar and hydrate to acidified water, pH 4.0 to 4.2
0.5% silver nitrate, 43 deg C, 2 min
Move to 58 deg C bath, 2 hr
DWW
Reducing solution, 58 deg C, 30-60 min
DWW
Return to same silver nitrate, 58 deg C, 10 min
DWW
Return to same reducing solution, 58 deg C, 5 min
DWw
NFR
DWW
DCM

Front 32

Churukian-Schenk results

Back 32

Arygrophil granules, argentaffin substances: black
Nuclei: red
Background: yellow-brown

Front 33

Citric acid, 0.3% solution

Back 33

Citric acid, 0.3 g
DW, 100 mL

Front 34

Acidified water

Back 34

DW, 100 mL
Add enough 0.3% citric acid to bring pH to 4.0 to 4.2

Front 35

Silver nitrate, 0.5% solution

Back 35

Silver nitrate, 0.5 g
DW, 100 mL

Front 36

Reducing solution, Churukian

Back 36

Sodium sulfite, anhydrous, 2.5 g
Hydroquinone, 0.5 g
DW, 50 mL
Prepare just before use

Front 37

GMS for urates protocol

Back 37

Depar to absolute alcohol, no 95 or water
Working methenamine silver solution, 60 deg C , 30 min or until urate crystals are black
DWW
3% hypo, 5 min
DWW
Working light green counter, 1.5-2 min
DCM

Front 38

GMS for urates results

Back 38

Urates: black
Background: blue-green

Front 39

Silver nitrate, 5% solution

Back 39

Silver nitrate, 25 g
DW, 500 mL

Front 40

Methenamine solution, 3%

Back 40

Hexamethylenetetramine, 3 g
DW, 100 mL

Front 41

Stock methenamine-silver nitrate solution

Back 41

Methenamine, 3% solution 100 mL
Silver nitrate, 5% solution, 5 mL
White precip will form but dissolve when shaken, store in fridge

Front 42

Sodium borate, 5% solution

Back 42

Sodium borate, 5 g
DW, 100 mL

Front 43

Working methenamine silver nitrate solution

Back 43

Methenamine silver solution, 25 mL
DW, 25 mL
Sodium borate, 5% solution, 3 mL

Front 44

3% hypo

Back 44

Sodium thiosulfate, 3 g
DW, 100 mL

Front 45

Light green stock solution

Back 45

Light green, SF yellowish, 1 g
DW, 500 mL
GAA, 1 mL

Front 46

Working light green solution

Back 46

Light green stock solution, 10 mL
DW, 50 mL

Front 47

Bile stain protocol

Back 47

Depar and hydrate to DW
Fouchet reagent, freshly filtered
DWW
van Gieson solution, 5 min
DCM

Front 48

Bile results

Back 48

Bile/bilirubin: emerald green to olive drab
Background: yellow

Front 49

Ferric chloride, 10% solution

Back 49

Ferric chloride, 10 g
DW, 100 mL

Front 50

Fouchet reagent

Back 50

Trichloracetic acid, 25 g
DW, 100 mL
Ferric chloride, 10 mL

Front 51

van Gieson solution

Back 51

Acid fuchsin, 1% solution 20 mL
Picric acid, saturated solution, 380 mL

Front 52

Von Kossa calcium stain protocol

Back 52

Depar and hydrate to DW
Silver nitrate solution, in glass coplin exposed to light: 30 min
DWW
5% hypo, 2 min
DWW
NFR, 2 min
DWW
DCM

Front 53

Von Kossa results

Back 53

Calcium salts: black
Background: red

Front 54

Alizarin red S calcium stain protocol

Back 54

Depar and hydrate to 50% alcohol
DWW, rapid
Alizarin red S staining solution, 30 sec-5 min
Shake off excess, blot
Acetone, 10-20 sec
Acetone-xylene, 10-20 sec
Xylene, mount

Front 55

Alizarin red S results

Back 55

Calcium deposits: orange-red

Front 56

Alizarin red S stain, 2% solution

Back 56

Alizarin red S, 2 g
DW, 100 ml
Mix, adjust pH to 4.1 to 4.3 with 0.5% ammonium hydroxide

Front 57

Rhodanine method for copper protocol

Back 57

Depar and hydrate to DW
Working rhodamine solution in plastic Coplin, 18 hr @ 37 deg C
DWW
Dilute Mayer hematoxylin, 10 min
DWW
0.5% sodium borate
DWW
DCM

Front 58

Rhodanine results

Back 58

Copper: bright red to red yellow
Nuclei: Light blue

Front 59

Saturated rhodanine solution

Back 59

5-(p-dimethylamino-benzylidine) rhodanine, 0.2 g
Absolute ethanol, 100 mL

Front 60

Working rhodanine solution

Back 60

Saturated rhodanine solution, 3 mL
DW, 47 mL
Filter before use

Front 61

Diluted Mayer hematoxylin

Back 61

Mayer hematoxylin, 25 mL
DW, 25 mL

Front 62

Sodium borate, 0.5%

Back 62

Sodium borate, 0.5 g
DW, 100 mL