THE UNIVERSITY OF CALIFORNIA AT RIVERSIDE *Standard Operating Procedure* *K. Le Roch Laboratory*
TITLE: GIEMSA OF BLOOD SMEARS AND ESTIMATION OF PARASITEMIA		Page 1 of 1
SOP #: KLR3	REVISION LEVEL:	DATE:

1 MATERIALS

1.1 Giemsa stain (LabChem Inc.)

1.2 100% methanol

1.3 Bibulous paper (VWR)

1.4 Microscope with ´100 oil immersion lens and 10´10 grid eyepiece

1.5 Microscope immersion oil

2 PROCEDURE

2.1 Giemsa staining of blood smeared slides

Fix slides in 100% methanol for ~30 s and rinse off in tap water;
Make up a fresh solution of 10% Giemsa stain in distilled water;
Stain ~30 min;
Rinse off slide in tap water and dry thoroughly using bibulous paper to dab.

2.2 Estimation of parasitemia

· View slide under oil immersion with a 100X objective;\

· Estimate parasitemia by counting the number of infected cells. A 10´10 grid square in the eyepiece of the microscope facilitates the procedure as an even blood smear yields ~100 red blood cells per 10´10 grid. Thus, for example, 8 infected blood cells in a 10´10 grid is 8% parasitemia. Several fields (~10) should be counted and the average taken to obtain a representative estimate of the total parasitemia.