Red blood cells (RBCs) are the most abundant type of blood cells, serving as the primary medium for oxygen transport through the blood in most animals, and also possess immune functions.
Dacie's Solution is composed of sodium citrate, formaldehyde, etc., and is an isotonic red blood cell diluent. The principle of Dacie's Solution is to dilute blood with an isotonic diluent at a certain ratio, charge the counting chamber, count the number of red blood cells within a defined volume under a microscope, and calculate the number of red blood cells per liter of blood.
This reagent is intended for research use only and is not suitable for clinical diagnosis or other applications.
Materials Required (Self-Prepared):
1.Fresh whole blood or EDTA-anticoagulated whole blood
2.Micropipette, hemocytometer (blood cell counting chamber), microscope
Operating Instructions (For Reference Only):
1.Take a medium-sized test tube and add 2.0 mL of Dacie's Solution.
2.Using a clean, dry micropipette, take 10 μL of peripheral blood or anticoagulated blood. Wipe off excess blood from the outside of the tube, then add it to the bottom of Dacie's Solution. Gently aspirate the upper layer of the solution to rinse the pipette 2–3 times, and mix immediately.
3.Using a clean micropipette, charge the red blood cell suspension into the counting chamber. Take care to avoid air bubbles or overflow. Let stand at room temperature for 2–3 min.
4.Under a microscope with high-power lens, count the red blood cells in the four corner squares and the central square (total of 5 medium squares) within the central large square. For cells touching the borderline, apply the principle of "count upper and left lines, do not count lower and right lines."
Calculation:
RBC count per liter = number of RBCs in 5 medium squares × 5 × 10 × 200 × 10⁶ = number of RBCs in 5 medium squares × 10¹⁰
