计算溶液所需的质量、体积或浓度。
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在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
红细胞(RBC)是血液中数量最多的一种血细胞,也是大多数动物体内通过血液运送氧气的媒介,同时还具有免疫功能。
Dacie氏液由柠檬酸钠、甲醛等组成,是一种等渗的红细胞稀释液。Dacie氏液作用原理是用等渗稀释液将血液按一定倍数稀释,充入计数池后显微镜下计数一定体积内红细胞数,换算求出每升血液中红细胞的数量。
该试剂仅用于科研领域,不适用于临床诊断或其他用途。
自备材料:
1、新鲜全血或EDTA抗凝全血
2、微量吸管、血细胞计数板、显微镜
操作步骤(仅供参考):
1、取中号试管,加Dacie氏液 2.0 mL。
2、用洁净干燥微量吸管取末梢血或抗凝血 10 μL,擦去管外余血后加至Dacie氏液底部,再轻吸上层清液清洗吸管 2~3 次,立即混匀。
3、用干净微量吸管将红细胞悬液充入计数池,注意避免产生气泡或外溢,室温静置 2~3 min。
4、置于显微镜高倍镜下依次计数中央大方格内四角和正中共5个中方格内的红细胞,压线细胞按“数上不数下,数左不数右”的原则进行计数。
计算:
红细胞数/L = 5个中方格内红细胞数 × 5 × 10 × 200 × 10⁶ = 5个中方格内红细胞数 × 10¹⁰
| ×5 | 5个中方格换算成1个大方格 |
| ×10 | 1个大方格容积为0.1 μL,换算成1.0 μL |
| ×200 | 血液的实际稀释倍数应为201倍,按200倍计算 |
| ×10⁶ | 由1 μL换算成1 L |
参考区间:
| 人群 | 参考值 (×10¹²/L) |
| 男 | (4.09~5.74) |
| 女 | (3.68~5.13) |
| 新生儿 | (5.2~6.4) |
| 婴儿 | (4.0~4.3) |
| 儿童 | (4.0~4.5) |
注意事项:
1、采血时不能过于挤压,针刺深度应适当。
2、不应以血红蛋白浓度来折算红细胞数。
3、试管、计数板均需清洁,以免杂质、微粒等被误认为红细胞。
4、在参考范围内,两次红细胞计数相差不应超过 5%。
5、试剂开封后请尽快使用,以防影响后续实验效果。
6、为了您的安全和健康,请穿实验服并戴一次性手套操作。
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