Iron
PRODUCT SPECIFICATIONS:
Tissue: Positive staining liver or spleen.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090).
Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.
Quality Control Stain: Gomori Prussian Blue quality control stained slide(s) included.
Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.
Storage: 15-30°C in a light deprived and humidity controlled environment.
Intended Use: To verify histological techniques and reagent reactivity.
Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.
CONTROL SLIDE VALIDATION:
With Iron, Gomori Prussian Blue Stain Kit: | Part 9136A/B | Individual Stain Solution | |
Solution A: | Hydrochloric Acid 20%, Aqueous | 125/250 ml | Part 12087 |
Solution B: | Potassium Ferrocyanide 10%, Aqueous | 125/250 ml | Part 13392 |
Solution C: | Nuclear Fast Red Stain, Kernechtrot | 250/500 ml | Part 1255 |
APPLICATION:
Newcomer Supply Iron Control Slides are for the positive histochemical staining of ferric iron deposits in tissue sections.
PRESTAINING PREPARATION:
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- Heat dry sections in oven according to your laboratory protocol.
- Acid clean glassware prior to use to avoid residual iron staining.
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- See Procedure Note #1.
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NEWCOMER SUPPLY VALIDATION PROCEDURE:
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- Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each. Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each. Wash well with distilled water.
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- See Procedure Notes #2 and #3.
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- Prepare fresh Ferrocyanide Working Solution directly before use; combine and mix well.
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- Solution A: Hydrochloric Acid 20%, Aqueous 20 ml
- Solution B: Potassium Ferrocyanide 10%, Aqueous 20 ml
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- Place slides in fresh Ferrocyanide Working Solution for 20 minutes.
- Rinse in three changes of tap water; rinse in distilled water.
- Place in Solution C: Nuclear Fast Red Stain, Kernechtrot for 5 minutes.
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- Shake solution well before use; do not filter.
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- Rinse well in distilled water.
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- See Procedure Note #4.
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- Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.
- Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each. Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each. Wash well with distilled water.
RESULTS:
Ferric iron deposits | Bright blue |
Nuclei | Red |
Cytoplasm | Pink |
PROCEDURE NOTES:
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- Acid clean all glassware/plasticware (Part 12086) and rinse thoroughly in several changes of distilled water.
- Drain slides after each step to prevent solution carry over.
- Do not allow sections to dry out at any point during procedure.
- Wash well after Nuclear Fast Red Stain, Kernechtrot to avoid cloudiness in dehydration steps.
- If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps
REFERENCES:
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- Luna, Lee G. Manual of Histologic Staining Methods of the Armed Forces Institute of Pathology. 3rd ed. New York: Blakiston Division, McGraw-Hill, 1968. 179-184.
- Sheehan, Dezna C., and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 217-218.
- Modifications developed by Newcomer Supply Laboratory.