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. Store at 15-30°C in a light deprived and humidity controlled environment.
These Basement Membrane control slides are produced from human surgical or autopsy tissues under carefully controlled conditions. Reasonable measures are used to deliver quality control slides that are as consistent as possible. However, characteristics of quality control slides may be dissimilar due to variations in the reagents, stains, techniques, laboratory conditions, and tissue sources used. At Newcomer Supply, we use a manual method of performing immunohistochemical and histopathology stains, specifically avoiding automation, in order to provide reactive control slides for even 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:
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
All glassware must be acid cleaned and thoroughly rinsed with distilled water.
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.
Prepare the Working Methenamine-Silver Solution:
Solution A, Methenamine, 3% Aqueous Solution 50 ml
Solution B, Silver Nitrate, 5% Aqueous Solution 2.5 ml
Solution C, Sodium Borate, 5% Aqueous Solution 5 ml
Heat the working silver solution to 45°C - 60°C. If precipitate forms in the working silver solution, the cause may be related to the solution temperature and the method used to heat the working silver solution.
Using non-metallic forceps and acid cleaned glassware, place slides in the preheated working silver solution. Incubate slides in oven, water bath, or on the bench top, (Set to the temperature which gives your lab the best results.) for approximately 12-18 minutes untill sections appear to be paper-bag brown. Periodically remove the control slide, rinse in warm distilled water, and check microscopically for adequate silver impregnation. Basement membranes should be dark brown. If the organisms are not sufficiently dark at this point, place slides back in the warm silver solution. Recheck at 2-3 minute intervals until desired intensity is achieved.
Rinse slides in 3-4 changes of distilled water.
Tone in Solution E, Gold Chloride 0.25% Aqueous Solution until sections turn grey. (20 seconds to 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|
|Nuclei||Outlined in Black|
- “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.