Marlene asks:
Does anyone have a reliable staining method to demonstrate aluminum in 
paraffin (not plastic) embedded tissues?
thank you in advance for the help-

Included are the methods I have used a number of years ago when dealing with
dementia in patients on long term dialysis.  The dementia was believed to be
due to aluminium, and in fact we did demonstrate it in a couple of cases.
I hope this helps.  Sorry, I don't have the references at my fingertips.
Leo Levesque
Pathology
Health Sciences Centre
Winnipeg, MB,Canada


ALUMINON STAIN FOR ALUMINUM HYDROXIDE
(Irwin, 1955)


For paraffin and celloidin sections.

SOLUTIONS

		Buffer Solution pH 5.2
				5M Aqueous ammonium chloride (267.5 gm/l)
44.2	parts
				5M Aqueous ammonium acetate (385.3 gm/l)
44.2	parts
				6M Aqueous hydrochloric acid (500 ml/l)	11.6
parts

		Aluminon Staining Solution
				Aurine tricarboxylic acid (Aluminon)	2
gm
				Buffer Solution pH 5.2	100	ml
				Dissolve the aurine tricarboxylic in a few
ml of the buffer and make up to 100 ml.  Heat to 75oC for 15 minutes and
filter while hot.

		Decolorizing Buffer Solution pH 7.2
				Buffer Solution pH 5.2	73.5	parts
				1.6M Aqueous ammonium carbonate solution
				(12.6 gm/100 ml)	26.5	parts
				Add one or the other of these solutions to
adjust to pH 7.2.

		Saturated Picric Acid Solution or 1/10,000 Methylene Blue
Solution


METHOD

1.	Bring sections down to water.
2.	Stain in Aluminon staining solution for 5 minutes at 75oC or 15
minutes at 60oC.
3.	Dip section three or four times in distilled water at room
temperature.
4.	Dip sections in decolorizing buffer solution for 3 seconds at room
temperature.  This decolorizes the pinkish-stained tissue, with little
effect on the color of the aluminum lake.  Longer treatment may diminish or
remove color of aluminum lake.
5.	Wash in water
6.	Counterstain with Picric Acid Solution or with a 1/10,000 Methylene
Blue Solution.
7.	Dehydrate, clear and mount.


RESULTS

		Aluminum hydroxide	- bright cherry red


Note:  If demonstrating aluminum in bone, undecalcified sections must be
used.
ALUMINON STAIN FOR ALUMINUM HYDROXIDE
(Irwin, 1955; HSC, 1983)


SOLUTIONS

		Buffer Solution pH 5.2
				5M Aqueous ammonium chloride (267.5 gm/l)
44.2	parts
				5M Aqueous ammonium acetate (385.3 gm/l)
44.2	parts
				6M Aqueous hydrochloric acid (500 ml/l)	11.6
parts

		Aluminon Staining Solution
				Aurine tricarboxylic acid (Aluminon)	2
gm
				Buffer Solution pH 5.2	100	ml
				Heat to 75oC for 15 minutes and filter while
hot.

		Decolorizing Buffer Solution pH 7.2
				Buffer Solution pH 5.2	73.5	parts
				1.6M Aqueous ammonium carbonate solution
				(12.6 gm/100 ml)	26.5	parts
				Add one or the other of these solutions to
adjust to pH 7.2.

		2% Metanil Yellow in Absolute Ethanol.


METHOD

1.	Bring sections down to water.
2.	Stain in Aluminon staining solution for 15 minutes at 60oC.
3.	Dip section three or four times in distilled water at room
temperature.
4.	Dip sections in decolorizing buffer solution for 3 seconds at room
temperature.  This decolorizes the pinkish-stained tissue, with little
effect on the color of the aluminum lake.  Longer treatment may diminish or
remove color of aluminum lake.
5.	Wash in water
6.	Counterstain with 2% Metanil yellow for 1 to 2 minutes.
7.	Dehydrate, clear and mount.


RESULTS

		Aluminum hydroxide	- bright cherry red


Note:  If demonstrating aluminum in bone, undecalcified sections must be
used.
Aluminon Stain for Aluminum
(Buchanan, 1981)


For Epon and Methacrylate sections.

Fixation:	Tissues should be fixed in 10% N.B. Formalin

Embedding:	Araldite Epon

Sections:	4 um


SOLUTIONS

		Buffer Solution pH 5.2
				5M Aqueous ammonium chloride (267.5 gm/l)
3.8	parts
				5M Aqueous ammonium acetate (385.3 gm/l)
3.8	parts
				6M Aqueous hydrochloric acid (500 ml/l)	1
part

		Aluminon Staining Solution
				Aurine tricarboxylic acid (Aluminon)	2
gm
				Buffer Solution pH 5.2	100	ml
				Dissolve the aurine tricarboxylic in a few
ml of the buffer and make up to 100 ml.  Heat to 75oC for 15 minutes and
filter while hot.

		Decolorizing Buffer Solution pH 7.2
				Buffer Solution pH 5.2	3.6	parts
				1.6M Aqueous ammonium carbonate solution
				(12.6 gm/100 ml)	10	parts
				Add one or the other of these solutions to
adjust to pH 7.2.

		Saturated Picric Acid Solution


METHOD


1.	Araldite Epon resin is not removed prior to staining.
2.	Stain in Aluminon staining solution for 30 minutes at 75oC.
3.	Wash in distilled water.
4.	Decolorise for 25 seconds in Buffer Solution pH 7.2
5.	Wash in distilled water
6.	Counterstain in Saturated Picric Acid Solution.
7.	Dry sections and mount.


RESULTS

		Mineralised bone	- rusty brown
		Osteoid and background	- yellow
		Aluminum hydroxide	- brilliant red