Jones' Method for Kidney

EMS26396

Components:

Periodic Acid 0.5% Aqueous

Methenamine Solution 3% Aqueous

Silver Nitrate 5% Aqueous

Borate Buffer Working Solution

Gold Chloride 0.2% Aqueous

Harris Haematoxylin

Alcoholic Eosin Y Stain Solution

Sodium Thiosulfate 3% Aqueous

Potassium Ferricyanide 0.5%

Sodium Metabisulfite 3% Aqueous

Acid Alcohol 1%

Ammonia Water 0.3%

Fixation:

10% Buffered Neutral Formalin or Russell’s Zenker.

Sections:

Cut paraffin at 2µm sections.

Staining Procedures:

1. Deparaffinise and hydrate slides to distilled water.
2. Oxidise in Periodic Acid, 0.5% for 11 minutes. Wash well in chloride-free water.
3. Prepare Methenamine Silver solution by mixing:
   1. 42.5 ml Methenamine 3%, 2.5ml Silver Nitrate, 5% and 12.0ml Borate Buffer, pH 8.2
4. Place slides in the solution and the entire jar in a water bath at 70°C for approx. 60 – 75 minutes. Check under microscope when slides appear medium brown microscopically. Every 10 minutes, once the medium brown color has been established, rinse slide in 70°C chloride-free water and check under microscope. Rinse again in hot water and return to hot staining solution. As the staining time approaches the end point, check the slides, as above, every 1 – 2 minutes. The entire procedure must be performed quickly to prevent an uneven staining of the tissue. The slides should exhibit a brownish-yellow background, intense black reticulum fibers, and black basement membranes. If the slides become oversaturated, i.e. too black, de-stain in a dilute Potassium Ferricyanide Solution, 0.5% for one or two dips.
5. Rinse well in distilled water. Tone in Gold Chloride, 0.2% for 1 minute. If sections are over-toned, place in Sodium Metabisulfite, 3% for 1 – 2 minutes. Rinse well in distilled water.
6. Place in Sodium Thiosulfate, 3% for 1- 2 minutes. Wash in running tap water, 10 minutes. Rinse well in distilled water.
7. Stain in Harris’ Haematoxylin containing 2 – 4 ml of Glacial Acetic Acid per 100 ml for 5 -15 minutes. Wash in water.
8. Differentiate in Acid Alcohol, 1% until the sections turned red.
9. Blue section in Ammonia Water, 0.3%. Wash thoroughly.
10. Counter-stain in Eosin Y, 1%, Alcoholic Solution
11. Dehydrate in 95% and 100% alcohol, and clear in Xylene, 3 changes each. Mount.

Stain Results:        

References:

Jones, D.B., Amer. J. Path. 27:99 (1951).

AFIP Manual of Histological Staining Methods, 3rd ed., Ed. L. Luna: NY, McGraw-Hill Publ., c. 1968, p.97.