Description
Hematoxylin is oxidized to hematein, which combines with a mordant to form a dye-metal complex. This complex selectively binds to nucleic acids within cell nuclei, producing blue to blue-purple nuclear staining following rinsing and/or bluing. Gill Hematoxylin I is formulated as a progressive stain, allowing nuclei to be stained directly without the differentiation step typically required for regressive hematoxylins.
VitroView™ Gill Hematoxylin I Solution is a ready-to-use, progressive nuclear stain intended for laboratory use in histology, cytology, and immunohistochemistry (IHC). The solution provides crisp nuclear staining with minimal background and is optimized for routine tissue and cytological specimen staining.
Gill Hematoxylin I is a single-strength formulation that produces light to moderate nuclear staining.
Application
- Progressive nuclear staining of tissue sections
- Hematoxylin and eosin (H&E) staining procedures
- Cytological specimen staining
- Counterstaining following immunohistochemical staining procedures
Kit Contents
VB-3050G1 VitroView™ Gill Hematoxylin I Solution —– 500 ml
Storage Condition
Room temperature.
Protocol
H&E 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).
- Place in Gill Hematoxylin I Solution for 1-3 minutes.
- Rinse in tap water for 5 min with 3 changes. Or Bluing reagent for 30–60 seconds.
- Rinse with water for 1 min.
- Place in 95% Ethanol for 1 minute (DO NOT RINSE).
- Place in Eosin for 1 min 30sec.
- Dehydrate with 2 changes of 95% Ethanol and 2 changes of 100% Ethanol (2 minutes per change).
- Clear with 3 changes of xylene (5 minutes per change)
- Mount coverslip onto glass slide with Permount or some other suitable organic mounting medium.
H&E Stain for Frozen Sections
- Frozen section are fixed in acetone at -20 °C for 3 min or 80% Methanol at 4°C for 5 min
- PBS x 10 min.
- Rinse in DI water (2 minutes).
- Follow step 3-10 above.
For Immunohistochemistry (IHC) Counterstain:
Following completion of chromogen development:
- Rinse slides thoroughly.
- Immerse in Gill Hematoxylin I for 30–90 seconds.
- Rinse with water.
- Blue nuclei.
- Dehydrate, clear, and mount.
Counterstaining time may be adjusted depending on tissue type and desired nuclear intensity.
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.
