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VitroView Jones Stain Kit (Basement Membrane) (#VB-3026)

 

Description

The periodic acid oxidizes the carbohydrate components of the basement membrane which produce aldehydes. The released aldehydes reduce the silver to visible metallic silver. With this principle, Jones Stain Kit (Basement Membrane) is intended for use in histological demonstration of the basement membrane and reticular fibers. This kit is ideal for staining renal glomerular basement membranes. The basement membrane and reticular fibers are also found in skin, spleen and lung, etc.

 

Kit Components

 

SKU#ReagentSize (ml)
VB-3026-1Periodic acid solution250
VB-3026-2Methenamine Solution250×2
VB-3026-3Silver Nitrate Solution30
VB-3026-4Borax Solution125
VB-3026-5Gold Chloride Solution125
VB-3026-6Sodium Thiosulfate Solution250
VB-3026-7Nuclear Fast Red Solution250

 

Storage

Store at 2-8°C

 

Protocol

Thing to do before staining

  • Equilibrate all materials and prepared reagents to room temperature just prior to use and gently agitate.
  • All glassware used in this procedure should be chemically cleaned and rinsed thoroughly in distilled water.
  • Do not use metal forceps to remove slides from reagents. Use plastic forceps only.

 

 Jones Stain for paraffin sections:

  • Deparaffinize in xylene I for 6 minutes and II for 6 minutes.
  • Rehydrate: ethanol 100% (2 minutes)x2; ethanol 95% (2 minutes)x2; ethanol 70% (2 minutes)x1; Rinse in distilled water (5 minutes).
  • Incubate slide in Periodic Acid Solution for 15 minutes.
  • Rinse in two changes of distilled water.
  •  Combine the following for a working Silver Methenamine Solution:
ComponentVolume
Methenamine Solution42 ml
Silver Nitrate Solution2.5 ml
Borax Solution6 ml

 

Δ Note: Mixed solution may not be stored for reuse later.

7.  Place working Silver Methenamine Solution in 50°C water bath and allow temperature to equilibrate.
8. Incubate slide in working Silver Methenamine Solution in 50°C water bath for 15 minutes. Using plastic forceps dip slide in ‘hot’ distilled water and check under a microscope for evaluation of silver impregnation. Basement membrane and reticular fibers should be black. If color is not sufficient, dip slide in ‘hot’ distilled water and return the slide to working Silver Methenamine Solution for 5-   10 minutes and check again.
9. Rinse in four changes of distilled water.
10. Incubate slide in Gold Chloride Solution for 15-30 seconds.
11. Rinse in four changes of distilled water.
12. Incubate slide in Sodium Thiosulfate Solution for 2 minutes.
13. Rinse in tap water followed by two changes of distilled water.
14. Incubate slide in Nuclear Fast Red Solution for 2 minutes.
15. Rinse slide quickly in distilled water.
16. Rinse slide using absolute alcohol.
17. Clear with 3 changes of xylene (5 minute per change)
18. Mount coverslip onto glass slide with Permount or some other suitable organic mounting medium.

 

Jones Stain for frozen sections:

  • Frozen section are fixed in acetone at -20 °C for 3 min or 80% Methanol at 4°C for 5 minutes
  • PBS 1× 10 minutes.
  • Rinse in DI water (2 minutes).
  • Follow step 3-12 above.

 

Expected Results:

  • Basement Membrane—————-Black
  • Reticulum Fibers———————–Black
  • Nuclei——————————————Red
  • Cytoplasm——————————-Light Pink

 

 Control Tissue: Kidney cut at 2-4 microns, lung or spleen section.

VitroView Jones Stain Kit (Basement Membrane) (#VB-3026)

SKU: VB-3026
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