INSTRUCCIONES DE USO

HAEMATOLOGICAL STAINS

Staining of human clinical specimens, for subsequent visualisation by microscopy.
For professional use only
Single use
Medical device for In Vitro Diagnostic

Intended use:
Haematological stains are staining reagents intended for use in cellular diagnostics in human medicine, in particular for the haematological and clinical-cytological examination of blood samples of human origin. The major part of the haematological staining methods are intended for stain human blood extensions to highlight haematopoyetic cells, chromosomes, parasites or other pathogens and are based on the use of Romanowsky stain. This stain has a mixture of eosine and methylene blue. Among these methods there are Wright, May-Grünwald, Giemsa and the fast staining technique.

General precaution:
• Not suitable for any other application than its intended use.
• Do not use if packaging is damaged.
• Do not use the product if it has changed its original color.
• Do not use if expiration date is overcome.
• Manage as hazardous waste.
• For its storage, keep the product closed at room temperature in a well-ventilated area and away from any hot or ignition points.

EOSIN-METHYLENE BLUE ACCORDING TO WRIGHT
Reagent:
Code: 808200 — Bottle of 250 ml
Instructions for use:
1. Take a clean grease-free slide.
2. Using the ground edge of a slide, thinly spread a drop of blood deposited on and near one end of another slide.
3. Dry the smear at room temperature.
4. Cover with 1 ml of stain during 1 minute for its action as a fixer.
5. Then add 1 ml water (pH 7.2).
6. Leave the stain to act for 2-4 minutes.
7. Rinse with water (pH 7.2).
8. Dry in a vertical position.
Note: As it is a fast staining, it is recommended to keep the smear as thin as possible.

EOSIN-METHYLENE BLUE ACCORDING TO MAY-GRÜNWALD
Reagents:
Code: 808000 — Bottle of 250 ml
Code: 808001 — Bottle of 1000 ml
Instructions for use:
1. Take a clean grease-free slide.
2. Using the ground edge of a slide, thinly spread a drop of blood deposited on and near one end of another slide.
3. On the smear dried at room temperature (not fixed), add 0,5 ml of May-Grünwald solution and let it to act during 2-3 minutes.
4. Add an equal amount of water (pH 7.2).
5. Mix carefully both solutions.
6. Leave for 5-10 minutes.
7. Rinse the slide and dry upright.

AZUR-EOSIN-METHYLENE BLUE ACCORDING TO GIEMSA
Reagent:
Code: 808100 — Bottle of 250 ml
Code: 808101 — Bottle of 1000 ml
Instructions for use:
1. Take a clean grease-free slide.
2. Using the ground edge of a slide, thinly spread a drop of blood deposited on and near one end of another slide.
3. Fix the smear with methanol and dry at room temperature.
4. Flush the slide with solution made of 10 drops of Giemsa solution in 10 ml water (pH 7.2).
5. Leave for 25 minutes.
6. Rinse the slide with water and dry upright.

FAST STAINING OF BLOOD SMEARS
Reagent:
Code: 805013 — Four bottles of 250 ml (Two bottles of A stain and two of B stain).
Instructions for use:
1. Prepare a thin, homogeneous blood smear in a slide.
2. Fix it with methanol for 2-3 minutes.
3. Immerse it in a container with Deltalab A stain (or totally cover the preparation).
4. Wait 5 seconds.
5. Drain the preparation in the edge of the container.
6. Rinse the smear in a container with clean wàter.
7. Let it drain.
8. Immerse the slide in Deltalab B stain (or totally cover the preparation).
9. Wait 10 seconds.
10. Drain the excess of stain.
11. Rinse in a second container with wàter.
12. Drain the excess of líquid and maintain it in an inclinated position until it is completely dried.
13. The extension must have a mauve colour, between blue and red
This method has the advantage that the laboratory can adapt the colour of the smear to
their preferences varying the time of the dye contact.
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Note: Dyes can be used several times, but is necessary to filter them every 2-4 days, depending on the number of extensions which are daily stained.
Results: In an extension correctly stain, the cells appear in the following way:
• Neutrophils: The cytoplasm will be dyed in pink colour and inside, the small granules will be dyed in mauve colour.
• Eosinophilis: The cytoplasm will be dyed in pink colour and inside, there can be observed big granules dyed in red colour.
• Monocytes: The cytoplasm will be dyed in grey colour.
• Lymphocites (big): The cytoplasm will be dyed in light blue colour.
• Lymphocites (small): The cytoplasm will be dyed in dark blue colour.
• Basophilis: Cell full of granules in dark purple colour.
• Erythocytes: Cells in light red colour.
• Platelets: Particles in purple-pink colour.

References:
1. John d. Bancroft, Marilyn Gamble. Theory and Practice of Histological Techniques, Churchill Livingstone Elsevier, Sixth Edition, 2008.
2. J. A. Kiernan et al. Histological & Histochemical Methods, Pergamon Press, Second Edition, 1990.
3. Paul Lopez Cardozo et al. Atlas of clinical cytology, EM edition medizing, 1968.
4. Horobin, R.W. and Kiernan, J.A. Conn’s Biological Stains: A Handbook of Dyes, Stains and Fluorochromes for Use in Biology and Medicine. Bios, 10th Edition, 2002.