BASEMENT MEMBRANE Control Slides
(Our product insert & how we do it)
BASEMENT MEMBRANE Control Slides
These quality control slides are intended to be used to verify histological techniques and reagent reactivity. They are to be used for the qualitative purpose of determining positive or negative results, and are not intended to be used for any quantitative purpose. The first serial section within this control box is stained and provided for your reference.
Newcomer Supply, Inc., takes reasonable measures to deliver quality control slides that are as consistent as possible. However, the characteristics of quality control slides may be dissimilar due to variations in the reagents, stains, techniques, laboratory conditions, and tissue sources used.
These basement membrane control slides were produced from human surgical or autopsy tissues. At Newcomer Supply we use a manual method of performing histopathology stains, specifically avoiding automation, in order to provide reactive control slides for even the less aggressive methods of staining that our customers may be using.
Fixation: 10% Neutral Buffered Formalin
Technique: Paraffin sections cut at 5 microns
Solutions: All solutions manufactured by Newcomer Supply
REAGENTS: (Available as a KIT or COMPONENTS)
KIT: Basement Membrane Stain Kit, Gomori, Catalog #9167
- Gomori Basement Membrane Silver Set: Catalog #1131
Solution A: Methenamine, 3%
Solution B: Silver Nitrate, 5%
Solution C: Sodium Borate, 5%
- Periodic Acid, 0.5% Aqueous Solution Catalog #13308
- Gold Chloride, 0.25% Aqueous Solution Catalog #11287
- Sodium Thiosulfate, 2.5% Aqueous Solution Catalog #13889
- Light Green SF Yellowish Stain Solution, 0.1% Aqueous Catalog #12203
STAINING PROCEDURE: Basement Membrane Stain, Gomori
- Prepare the Working Methenamine-Silver Solution as follows:
- Solution A, Methenamine, 3% Aqueous Solution 50ml
- Solution B, Silver Nitrate, 5% Aqueous Solution 2.5ml
- Solution C, Sodium Borate, 5% Aqueous Solution 5ml
Preheat this working solution along with a coplin jar of distilled water in a 60°C water bath.
- Deparaffinize sections thoroughly in 3 changes of xylene, then hydrate through graded ethyl alcohols to distilled water.
- Place slides in Solution D, Periodic Acid, 0.5% Aqueous Solution for 15 minutes.
- Wash slides in gently running tap water for 5 minutes.
- Rinse slides well in distilled water.
- Using non-metallic forceps, place the slides in the preheated working Methenamine-Silver Solution in the 60°C water bath until the sections on the slide begin to turn a light to medium brown. This should take approximately 10-15 minutes.
- Remove the control slide from the Methenamine-Silver Solution and place in the heated distilled water. Check under a microscope. The basement membrane should be black with some cytoplasm light to Medium brown. If the basement membrane isn’t intense enough, rinse in heated distilled water and place back in the Methenamine-Silver solution. Check every 1-2 minutes until desired intensity is achieved.
- Rinse all slides in 3 changes of distilled water.
- Tone sections in Solution E, Gold Chloride 0.25% Aqueous Solution for 1 minute.
- Rinse slides well in 3 changes of distilled water.
- Place in Solution F, Sodium Thiosulfate 2.5% Aqueous Solution for 2 minutes.
- Wash slides in gently running tap water for 5 minutes, followed by a distilled water rinse.
- Place slides in Solution G, Light Green SF Yellowish Stain Solution, 0.1% for 1 minute.
- Quickly rinse slides in distilled water, 2 dips.
- Dehydrate through graded ethyl alcohols, clear in xylene or substitute, and coverslip with a compatible mounting medium.
- Kidney Glomerular Basement Membranes: Black
- Intraglomerular Deposits: Black
- Nuclei: Outlined in Black
- Cytoplasm: Light Green
- “Manual of Histologic Staining Methods of the Armed Forces Institute of Pathology”, Luna, L., 3rd Ed., 1968, pp. 97-99.
- “Theory and Practice of Histotechnology”, Sheehan, D. & Hrapchak, B., 2nd Ed., 1980, pp. 187-188.
- Modifications developed by Newcomer Supply Laboratory.