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红细胞（Red blood cell，RBC）也称红血球，是血液中数量最多的一种血细胞，脊椎动物体内通过RBC运送氧气，RBC同时还具有免疫功能，在贫血检查中可通过红细胞渗透脆性试验来检测红细胞膜的缺陷。
检测原理：红细胞悬浮于等渗盐水中能够保持双面凹盘状态，如果渗透压增高，红细胞水分会渗出细胞外而呈现皱缩，如果渗透压降低，则水分会渗入细胞内，使红细胞膨胀以致破裂、溶血，利用这一原理将红细胞加至一系列的不同浓度的低渗盐水中，检查溶血程度，以便判断红细胞抵抗低渗溶液的能力，该实验被称为红细胞渗透脆性试验，主要用于检测人、动物血液的红细胞渗透脆性。该试剂盒仅用于科研领域，不适用于临床诊断或其他用途。
自备仪器和试剂
小试管、分光光度计
操作步骤
1、配制Parpart NaCl工作液
按照Parpart NaCl Solution：ddH₂O=1：9的比例配制适量的Parpart NaCl工作液。
2、样本制备
无菌采血，制成肝素抗凝血或脱纤维血，充分混匀待用。
3、取小试管或恰当容器，编号并按照Parpart NaCl稀释表依次加入Parpart NaCl工作液和ddH₂O，即为Parpart NaCl稀释液，其总体积为4ml，每次检查应设正常对照。
4、分别吸取40μl肝素抗凝血或脱纤维血加入上述Parpart NaCl稀释液中，立即混匀，室温（20～30℃）静置30min。
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6、结果计算与参考区间
溶血百分率%=各管吸光值/100%溶血管吸光值×100
参考区间：50%溶血率 4.0～4.45g/L
注意事项
1、每次检查应设正常对照，正常对照与待测样品氯化钠浓度相差0.4g/L，即有诊断意义。
2、Parpart NaCl Solution采用高纯度氯化钠配制，不要被酸碱污染，同时注意密闭保存。
3、血液样品应直接滴入液体，不宜沿管壁流入。
4、不能用枸橼酸盐或双草酸盐作抗凝剂，以免增加离子强度，影响溶液的渗透压。
5、试剂开封后请尽快使用，以防影响后续实验效果。
Red Blood Cells (RBCs), also known as erythrocytes, are the most abundant type of blood cell in the blood. In vertebrates, RBCs transport oxygen throughout the body and also possess immune functions. In anemia testing, defects in the red blood cell membrane can be detected through the red blood cell osmotic fragility test.
Test Principle:
When suspended in an isotonic saline solution, red blood cells maintain their biconcave disc shape. If the osmotic pressure increases, water will flow out of the cells, causing them to shrink. Conversely, if the osmotic pressure decreases, water will enter the cells, causing them to swell and eventually rupture, leading to hemolysis. Based on this principle, red blood cells are added to a series of low‑osmotic saline solutions of different concentrations, and the degree of hemolysis is examined to determine the resistance of the red blood cells to hypotonic solutions. This test is referred to as the red blood cell osmotic fragility test and is primarily used to assess the osmotic fragility of human and animal blood cells. This kit is intended for research use only and is not suitable for clinical diagnosis or other purposes.
Reagents, consumables and Equipments not provided
Small test tubes, spectrophotometer
Procedure
1. Preparation of Parpart NaCl Working Solution
Prepare an appropriate amount of Parpart NaCl working solution by mixing Parpart NaCl Solution and ddH₂O at a ratio of 1:9.
2. Sample Preparation
Collect blood aseptically and prepare heparin‑anticoagulated blood or defibrinated blood. Mix thoroughly before use.
3. Dilution Series
Take small test tubes or appropriate containers, label them, and sequentially add Parpart NaCl working solution and ddH₂O according to the Parpart NaCl dilution table. The total volume per tube should be 4 mL. A normal control should be included in each assay.
4. Adding Blood Samples
Add 40 μL of heparin‑anticoagulated blood or defibrinated blood to each of the above Parpart NaCl dilution tubes. Mix immediately and let stand at room temperature (20–30 °C) for 30 minutes.
5. Measurement
Centrifuge each tube for 5 minutes and take the supernatant. Measure the absorbance at 540 nm or using a green filter on a spectrophotometer. Use the supernatant from the tube containing 8.5 g/L NaCl (tube No. 3) as the blank to zero the instrument. Use the tube containing 1 g/L NaCl (tube No. 17, or a tube in which ddH₂O replaces the Parpart NaCl dilution solution and 40 μL of blood is added) as the 100% hemolysis tube. Calculate the percentage of hemolysis for each tube. Report the result as the NaCl concentration that produces 50% hemolysis.
Table 1: Parpart NaCl Dilution Table and Hemolysis Rate
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6. Calculation of Results and Reference Range
Hemolysis Percentage (%) = (Absorbance of each tube / Absorbance of 100% hemolysis tube) × 100
Reference Range: 50% hemolysis concentration – 4.0 to 4.45 g/L
Notes
1. Each assay should include a normal control. A difference of 0.4 g/L in NaCl concentration between the normal control and the test sample is considered diagnostically significant.
2. Parpart NaCl Solution is prepared from high‑purity sodium chloride. Avoid contamination by acids or alkalis and store tightly sealed.
3. Blood samples should be added directly into the liquid; do not allow them to flow down the tube wall.
4. Do not use citrate or double oxalate as anticoagulants, as they may increase ionic strength and affect the osmotic pressure of the solution.
5. Use the reagents as soon as possible after opening to avoid affecting subsequent experimental results.
| R1507805 | Component | 100T | Storage |
| R1507805A | Parpart NaCl Solution | 30 mL | RT. |
| R1507805B | ddH₂O | 500 mL | RT. |
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 ZJ26F0131497 |
分析证书 | R1507805 |
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