Description
Hematoxylin is oxidized to hematein, which combines with an aluminum mordant to form a dye-metal complex. This complex selectively binds to nuclear chromatin and nucleic acids. Following rinsing and bluing, nuclei appear blue to blue-purple with excellent chromatin definition.
Gill Hematoxylin III is a high-concentration formulation that enables rapid progressive staining. Depending on staining time, it may also be used in regressive protocols requiring differentiation.
VitroView™ Gill Hematoxylin III Solution is a ready-to-use, triple-strength progressive nuclear stain formulated for histology, cytology, frozen section diagnostics, and immunohistochemistry (IHC). It provides rapid, intense blue to blue-purple nuclear staining with excellent chromatin detail and minimal background.
Gill Hematoxylin III is the strongest formulation in the Gill series and is preferred when darker nuclear staining or shorter staining times are required.
Application
- Progressive nuclear staining of formalin-fixed, paraffin-embedded (FFPE) tissue sections
- Routine Hematoxylin and Eosin (H&E) staining
- Cytology staining where increased nuclear intensity is desired
- Counterstaining in immunohistochemistry (IHC)
Kit Contents
VB-3050G3 VitroView™ Gill Hematoxylin III 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 III Solution for 1-2 minutes (adjust according to tissue type and desired intensity).
- 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 20–60 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.
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User Manual and Material Safety Data Sheet (MSDS) (PDF )
VB-3050G3 MSDS
