Jones

Author: Kelsey Goldman, HTL (ASCP)

Last Update: 11 July 2023

Cite this Page! -> Goldman, Kelsey. “Jones.” HistologyOutlines.Com, 11 July 2023. Accessed (today)

Purpose:

An argyrophilic stain for the visualization of basement membranes; primarily used in the staining of kidney sections. This stain is also called the Jones Methenamine Silver Method (Colangelo).

Control Tissues:

Staining Pattern:

Biochemistry Theory:

The Jones staining method is an example of argyrophilic staining. This meaning that the tissue itself will act to reduce the silver from its ionic form to the visible metallic form; there is no need to add a secondary reducing agent. In the case of the Jones stain, the silver reduction takes place with proteins found on the thick surface of the kidney’s basement membranes (Colangelo).

Troubleshooting:

Always use chemically cleaned glassware with silver staining and avoid the use of metallic forceps (Luna)
As noted in the protocol below, it is very important to carefully follow heating and filtering steps. The misuse of these steps may lead to grainy particulate on the slide.

Additional References:

Colangelo, Gail. “Basement Membrane Staining in Renal Biopsies.” Journal of Histotechnology, vol. 1, no. 2, 1977, pp. 63–64, https://doi.org/10.1179/his.1977.1.2.63.
Luna, Lee. Manual of Histologic Staining Methods of the Armed Forces Institute of Pathology. Third, McGraw Hill, 1968.

Protocols

Luna's Adaptation of the Jones Stain

Solutions:

0.5% Periodic Acid
3% Methenamine ( can be diluted from stringer or powered solution in distilled water)
5% Silver nitrate
Borate Buffer stock A ( 0.2 M Boric Acid )
- Boric Acid – 12.36 g
- distilled water 1000 mL
Borate Buffer Stock B (0.25 M Sodium Borate)
- Sodium Borate 19.07 g
- Distilled water 1000 mL
Working Borate Solution pH 8.2
- Solution A 6.5 mL
- Solution B 3.5 mL
Gold Chloride ( replace after 100 slides)
3% sodium Thiosulfate
Working Methenamine silver pH 8.2 Solution
- Methenamine 3%- 42.5 mL
- Silver nitrate 5%- 2.5 mL
- Borate Buffer pH 8.2, 12 mL
- Filter and prepare fresh , Stable approximately 1 hour

Protocol

Deparaffinize and hydrate slides
Incubate in the periodic acid solution for 11 minutes
wash in distilled water
Place slides in the methenamine – silver working solution and place into a 70C water bath for 60-75 minutes. Do not forget to pre-filter this solution. Do not allow slides to go over temperature. Check under the microscope to ensure that a brown “paper bag” color is present on the slide. Do not dry out the slide in the process of checking it. Try to make sure the slide doesn’t cool in this process; this will lead to uneven staining of the section.
rinse well in distilled water
Tone in the gold chloride solution for 1 minute
rinse in distilled water
incubate in Sodium thiosulfate for 1-2 minutes.
wash in running tap water for 10 minutes
Counterstain the sections in Harris hematoxylin and eosin
dehydrate, clear, and coverslip

Adapted from

Luna, Lee. Manual of Histologic Staining Methods of the Armed Forces Institute of Pathology. Third, McGraw Hill, 1968.