The Modified Hematoxylin-Eosin (HE) Staining Kit is a core tool for routine staining of pathological sections. It features convenient operation and stable staining effects, and is widely used in histological morphology observation. Primarily designed for morphological staining of biological tissue sections (such as paraffin sections and frozen sections), it can clearly distinguish cell nuclei from cytoplasm, aiding in the observation of tissue structure, cell morphology, and pathological changes. It serves as the most fundamental staining method in pathological diagnosis and scientific research experiments.
H1506002
| Components
| 100T
| Storage
|
H1506002A
| Hematoxylin Staining
| 100 mL
| RT. Store in the dark.
|
H1506002B
| Eosin Staining
| 100 mL
| RT
|
H1506002C
| Blueing Reagent
| 100 mL
| RT
|
H1506002D
| Differentiating Solution
| 100 mL
| RT
|
Self-Prepared Materials:
1. 1% acidic ethanol differentiating solution; alternatively, acidic ethanol differentiating solution can be purchased.
2. Weakly alkaline water; alternatively, ammonia solution (0.1%) can be purchased.
3. Lithium carbonate aqueous solution; alternatively, lithium carbonate aqueous solution (1%) can be purchased.
4. A series of ethanol solutions, including absolute ethanol, 95% ethanol, 90% ethanol, and 80% ethanol.
5. 4% paraformaldehyde; alternatively, paraformaldehyde solution (4% PFA) can be purchased.
Operating Procedures (For Reference Only):
Staining of Paraffin Sections:
1. Dewaxing of Sections to Water
① Treat with xylene twice, 5–10 minutes each time.
② (Optional) Treat with absolute ethanol twice, 3–5 minutes each time.
③ Treat with 95% ethanol for 3–5 minutes.
④ Treat with 90% ethanol for 3–5 minutes.
⑤ Treat with 80% ethanol for 3–5 minutes.
⑥ Rinse with tap water or distilled water for 1–3 minutes.
2. Staining
① Stain with hematoxylin staining solution for 5–8 minutes.
② Rinse with distilled water for 5–10 seconds to remove excess staining solution.
③ Immerse the sections in differentiation solution for 2–5 seconds, then rinse thoroughly with running tap water.
④ Treat the sections with bluing solution for 2–5 seconds, then rinse thoroughly with running tap water.
⑤ Stain with eosin staining solution for 3–5 minutes.
3. Dehydration, Clearing, and Mounting
① Treat with 80% ethanol for 10–20 seconds.
② Treat with 90% ethanol for 10–20 seconds.
③ Treat with 95% ethanol twice, 1–2 minutes each time.
④ Treat with absolute ethanol twice, 2–3 minutes each time.
⑤ Clear with xylene three times, 2–3 minutes each time.
⑥ Mount sections with neutral resin.
Staining Results
Cell nuclei: blue.
Cytoplasm, muscle fibers, collagen fibers, and thyroid colloid: red of varying intensities.
Keratin and red blood cells: bright orange-red.
Staining of Frozen Sections
① Fix with acetone or paraformaldehyde for 5–10 seconds.
② Rinse with tap water for 2–5 seconds.
③ Drop-stain with hematoxylin staining solution for 1–2 minutes (may be heated to 50°C).
④ Rinse with tap water for 2–5 seconds.
⑤ Immerse the sections in differentiation solution for 2–5 seconds, then rinse thoroughly with running tap water.
⑥ Treat the sections with bluing solution for 2–5 seconds, then rinse thoroughly with running tap water.
⑦ Stain with eosin staining solution for 2–5 seconds.
⑧ Rinse with water for 1–2 seconds.
⑨ Treat with 80% ethanol for 1–2 seconds.
⑩ Treat with 95% ethanol for 1–2 seconds.
⑪ Treat with absolute ethanol for 2–5 seconds.
⑫ Treat with phenol-xylene (1:3, v/v) for 2–5 seconds.
⑬ Clear with xylene for 3 times, 2–5 seconds each time.
⑭ Mount the sections with neutral balsam.
Precautions:
1. Paraffin sections should be thoroughly deparaffinized.
2. The 95% ethanol, 90% ethanol, and 80% ethanol solutions should be replaced regularly with fresh solutions.
3. The differentiation time in 1% acid-alcohol differentiation solution should be determined based on the thickness of the sections, the type of tissue, and the freshness of the 1% acid-alcohol differentiation solution. Additionally, after differentiation, the sections should be rinsed sufficiently with running water to completely remove the acid.
4. The ether-ethanol fixation solution is prepared by mixing equal amounts of ether and 95% ethanol, followed by adding an appropriate amount of acetic acid. It should be stored in a sealed container.
5. The staining time for frozen sections should be as short as possible.
6. Bluing solutions commonly used include 0.2–1% ammonia water, Scoot bluing solution, or 0.1–1% lithium carbonate aqueous solution.
7. For your safety and health, please wear a lab coat and disposable gloves during operations.
