VitroView™ Jones Stain Kit (Basement Membrane)

$320.00

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SKU: VB-3026 Category:

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# Reagent Size (ml)
VB-3026-1 Periodic acid solution 250
VB-3026-2 Methenamine Solution 250×2
VB-3026-3 Silver Nitrate Solution 30
VB-3026-4 Borax Solution 125
VB-3026-5 Gold Chloride Solution 125
VB-3026-6 Sodium Thiosulfate Solution 125
VB-3026-7 Nuclear Fast Red Solution 125

Storage

Store at 2-8°C for VB-3026-1/2/3/5 and room temperature for VB-2026-4/6/7

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:

  1. Deparaffinize in xylene I for 6 minutes and II for 6 minutes.
  2. Rehydrate: ethanol 100% (2 minutes)x2; ethanol 95% (2 minutes)x2; ethanol 70% (2 minutes)x1; Rinse in distilled water (5 minutes).
  3. Incubate slide in Periodic Acid Solution for 15 minutes.
  4. Rinse in two changes of distilled water.
  5.  Combine the following for a working Silver Methenamine Solution:
Component Volume
Methenamine Solution 42 ml
Silver Nitrate Solution 2.5 ml
Borax Solution 6 ml

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

7.  Place working Silver Methenamine Solution in 65°C water bath and allow temperature to equilibrate.
8. Incubate slide in working Silver Methenamine Solution for 60 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:

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

Note

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

Precautions

Handle with care. Avoid contact with eyes, skin and clothing. Do not ingest. Wear gloves.

User Manual and Material Safety Data Sheet (MSDS)  (PDF)

VB-3026 User Manual

VB-3026 MSDS