In a narrow sense, connective tissue refers to the three types of fibers it contains: collagen fibers, reticular fibers, and elastic fibers. There are many staining methods for connective tissue, such as Masson trichrome staining, Van Gieson staining, Gomori ammoniacal silver method, and Mallory phosphotungstic acid hematoxylin staining. However, the above methods only focus on the staining of one or two kinds of tissues.
Russell Modified Movat Pentachrome Staining Solution is widely used for its rich and bright staining. This method is mainly used to demonstrate atherosclerotic plaques. Weigert hematoxylin is used to stain cell nuclei, safranine fuchsin to stain cytoplasm, crocein scarlet to stain collagen tissue, and alcian blue to stain the matrix (proteoglycans). Due to the complicated operation procedure of this kit, the staining effect is highly related to the experience and proficiency of the operator, so it is not easy to obtain consistently satisfactory results. This reagent is for research use only. Not for clinical diagnosis or other purposes.
Product Components and Storage Conditions:
R1518553
| Component
| 9×50mL
| Storage
|
R1518553A
| Hypo Solution
| 50mL
| RT
|
R1518553B
| Alcian Blue Staining Solution
| 50mL
| 2-8℃. Store in the dark.
|
R1518553C
| Alkalic Alcohol Solution
| 50mL
| RT
|
R1518553D
| Weigert A
| 30mL
| RT
|
R1518553E
| Weigert B
| 20mL
| RT
|
R1518553F
| Weigert C
| 10mL
| RT. Store in the dark.
|
R1518553G
| Safranin Fuchsin A
| 40mL
| RT
|
R1518553H
| Safranin Fuchsin B
| 10mL
| RT
|
R1518553I
| Phosphotungstic Acid Solution
| 50mL
| RT. Store in the dark.
|
R1518553J
| Weak Acid Differentiation Solution
| 50mL
| RT
|
R1518553K
| Alcoholic Crocein Scarlet Staining Solution
| 50mL
| RT
|
R1518553L
| Bouin Fixative
| 250mL
| RT. Store in the dark.
|
Immediately before use, mix D:E:F at a ratio of 3:2:1 to obtain Weigert hematoxylin. Do not prepare in advance.
Immediately before use, mix G:H at a ratio of 4:1 to obtain safranin fuchsin staining solution. Do not prepare in advance.
Materials Required:
1. Graded ethanol, distilled water, xylene or eco-friendly dewaxing and clearing agent, neutral balsam
2. Microwave oven
Protocol (For Reference Only):
1. Deparaffinize paraffin sections routinely with xylene or dewaxing and clearing agent, and rehydrate with graded ethanol.
2. Place an appropriate amount of Bouin fixative in a microwave oven, heat moderately for 30–60 seconds, then immediately immerse the sections for 10 minutes.
3. Rinse in running water for 10 minutes.
4. Treat sections in Hypo Solution for 5 minutes, then rinse 2–3 times with distilled water.
5. Stain sections in Alcian Blue Staining Solution for 20 minutes, then rinse in running water for 2–5 minutes.
6. Preheat Alkalic Alcohol Solution in a microwave oven at high power to 45–60 °C, then immerse sections for 10 minutes.
7. Rinse in running water for 2–5 minutes.
8. Stain sections in pre-mixed Weigert Hematoxylin Staining Solution for 60 minutes, protected from light.
9. Rinse briefly in running water, then wash 2–3 times with distilled water, 3–5 minutes each time.
10. Stain sections in pre-mixed Safranin Fuchsin Staining Solution for 1 minute, protected from light.
11. Rinse 2–3 times with distilled water, 3–5 minutes each time.
12. Treat sections in Phosphotungstic Acid Solution for 5 minutes, then transfer directly into Weak Acid Differentiation Solution for 5 minutes.
13. Rinse 2–3 times with distilled water, 3–5 minutes each time.
14. Dehydrate: 95% ethanol for 1 minute, 100% ethanol twice for 1 minute each.
15. Stain sections in Crocein Scarlet Staining Solution for 5 minutes.
16. Dehydrate: absolute ethanol twice for 1 minute each, clear in xylene or dewaxing and clearing agent, and coverslip.
Staining Results:
Nuclei and elastic fibers: Black
Collagen and reticular fibers: Yellow
Proteoglycans: bluish Green
Fibrinoid, deep cellulose: Deep red
Cardiac muscle and smooth muscle: Red
Foam cells: Purple
Precautions:
1. Due to staining intensity, tissue section quality and other factors, not all five colors may be displayed after staining. Anti-slip section treatment is recommended.
2. Section thickness is generally about 5 μm; elastic fiber differentiation is usually completed within 2–3 minutes.
3. Thorough rinsing in running water to remove Weigert Hematoxylin is critical; inadequate rinsing will inhibit subsequent staining steps.
4. This staining method can demonstrate Cryptococcus neoformans, which is stained bright blue.
5. Use reagents promptly after opening to avoid affecting experimental results.
6. For your safety and health, wear a lab coat and disposable gloves during operation.
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