SN/T 1678-2005Piroplasmosis pathogen identification method (English PDF)

2.1 Reagents

---EDTA solution (1mg/mL) (see Appendix A);

---5% and 10% of Giemsa dye solution (see Appendix A);

---Methanol (analytical grade).

2.2 Instruments

---Optical microscope (8 times or more eyepieces and 60 times more objective lenses);

--- Eyepiece micrometer;

---Mirror micrometer;

--5mL syringe, 20 gauge needle;

---Slides.

4.1 Smear making and fixing

4.1.1 Production of thin blood film

Drop 1 drop of blood on the left end of the clean glass slide, take a small amount of blood with a pusher, and apply a thin blood film from the near right end of the slide. After air drying, Fix with methanol for 1min, then dye with 10% Giemsa dye solution for 20min~30min, just apply the blood film as soon as possible to ensure good the quality of the dyeing.

4.1.2 Production of thick blood film

Drop 1 drop of blood (about 50 µL) on a clean glass slide, heat it at 80 ° C for 5 min, then use 5% Giemsa dye solution.

Dyeing for 20min to 30min.

4.1.3 Production of organ smear

Lightly touch the fresh cut surface of the organ with a clean glass slide, or gently clamp a small piece of tissue with 2 clean slides, and push the glass vertically the piece makes the two slides leave a layer of tissue. After the smear is air-dried, it is fixed with methanol for 5 min, and then dyed with 10% Giemsa dye solution for 20 min~ 30min.

4.1.4 Preparation of lymph smears

The lymph fluid extracted by the syringe is pushed onto the slide glass, and then smeared with a needle to prepare a smear. After the smear is air-dried, it is fixed with methanol.