How to stain a smear for piroplasmosis in dogs? - briefly
To stain a smear for piroplasmosis in dogs, use Giemsa or Wright-Giemsa stains, which effectively highlight the parasites for microscopic examination. Begin by fixing the smear with methanol before applying the stain and following standard staining protocols to ensure optimal visualization of the organisms.
How to stain a smear for piroplasmosis in dogs? - in detail
To effectively stain a smear for the diagnosis of piroplasmosis in dogs, follow these detailed steps:
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Sample Collection: Begin by collecting a small amount of blood from the dog's ear or tail vein. Use a capillary tube or a fine needle to draw blood directly onto a glass slide. This will create a thin smear, which is ideal for staining and microscopic examination.
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Slide Preparation: Allow the blood smear to air dry completely. Do not use heat, as it can distort the cells and make identification of piroplasms more difficult. Once dried, fix the slide by immersing it in methanol for 3-5 minutes. This step helps to preserve the cellular structure and enhances staining quality.
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Staining Procedure: Choose an appropriate stain for detecting piroplasms. Commonly used stains include Wright's stain, Giemsa stain, or a Romanowsky-type stain. Prepare the stain according to the manufacturer’s instructions. Typically, this involves mixing powdered stain with methanol and distilled water.
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Staining Process:
- Submerge the fixed slide in the staining solution for 20-30 minutes. Ensure that the entire smear is covered by the stain.
- After staining, rinse the slide gently with tap water to remove excess stain. Be careful not to disturb the cells on the slide.
- Allow the slide to air dry completely. This step ensures that the stain sets properly and prevents smudging under the microscope.
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Microscopic Examination: Once the slide is dry, examine it under a microscope using oil immersion objectives (100x). Piroplasms appear as small, round to oval bodies within red blood cells. They often have a distinctive blue or purple color depending on the stain used.
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Interpretation: Identify and count the number of piroplasms in infected red blood cells. A positive diagnosis is typically based on the presence of at least one piroplasm per 100 high-power fields (HPFs). Document your findings, noting the number and distribution of piroplasms observed.
By following these detailed steps, you can effectively stain a smear for the diagnosis of piroplasmosis in dogs, ensuring accurate and reliable results.