University:
University of Cincinnati Blue Ash
Course:
BIOL 5180 | Immunology
Academic year:

2024
Views:

83

Pages:

2
Author:

Jasper J.

Giemsa Staining in Thin and Thick Blood Smears 1. Giemsa Staining of Thin Blood Smears 1. Make sure the preparation is completely dry 2. Thin blood smears are then fixed with absolute methanol The position of the slide is slightly tilted when dripping methanol or the slide is dipped in methanol for a short time (1-2 seconds) with the thin blood at the bottom, because the methanol should not touch the thick blood (it interferes with the dehemoglobinization of the thick blood smear) 3. Let the thin blood dry from methanol for ± 1 minute. 4. Prepare Giemsa liquid which has been made with a 3% dilution. 5. Place the slide on the staining rack and apply Giemsa liquid using a pipette until all the blood smear is covered in paint. 6. Leave it for 45-60 minutes 7. Rinse slowly and carefully with clean water 8. Let the slides dry in a standing position 2. Giemsa Staining of Thick Blood Smears Procedure: 1. Prepare a Giemsa dye solution by diluting the dye: - 5% solution for 30 minutes staining, add 1.5 ml of Giemsa dye to 50 ml of 7.1-7.2 saline buffer and mix well10% solution for 10 minute staining, add 5 ml of Giemsa dye to 45 ml of 7.1-7.2 saline buffer and mix well 2. Place the slide on a staining rack orcopy jar, crossing the stem of the glass pipe 3. Pour the dye solution provided into itCoplain jaron the slide (if using a rack), so that it covers the entire surface of the preparation 4. Leave for 30 minutes for a 5% dye solution or 10 minutes for a 10% dye solution 5. Wash in buffered water. (Do not remove Giemsa dye before washing in buffered water because it will leave dye deposits on top of the smear.) 6. Remove and clean the bottom of the slide, place it on a drying rack, until dry 7. Take the Giemsa stain preparation and place it on the microscope object table, drop 1 drop of immersion oil on top of the preparation 8. Check with a 40x objective lens and 10x eyepiece, then replace with magnification. Look for a parasite that has a clear nucleus and cytoplasm (red and blue). For older stages, pigments that are light brown or even black can help. If plasmodium is known, the next step is to estimate the possibility of the presence of other types of plasmodium 9. Carry out the inspection in a zig-zag manner, namely from one side to the other, then back to the original side and so on