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台盼蓝染色试剂盒(Trypan Blue Dyeing Kit)用于细胞培养实验中活细胞，死细胞计数以及细胞存活率测定实验。台盼蓝是细胞活性染料，常用于检测细胞膜的完整性与细胞的存活率，是组织和细胞培养中最常用的死细胞鉴定染色方法之一。正常的活细胞细胞膜结构完整，能够排斥台盼蓝，细胞不会被染成蓝色；而丧失活性或细胞膜不完整的细胞可被台盼蓝染成蓝色。通常认为细胞膜完整性丧失，即可认为细胞已经死亡。因此，台盼蓝染色后，通过显微镜下观察颜色的差异即可肉眼区分死细胞和活细胞，通过计数就可对细胞存活率进行比较精确的计算。
使用方法
1. 收集细胞：
如果收集贴壁细胞，应用胰蛋白酶-EDTA溶液消化细胞，收集细胞。如果收集悬浮细胞，则可以直接收集细胞。收集好的细胞在1000rpm离心5min，弃上清，用1mL细胞重悬液重新悬浮细胞沉淀。
2. 台盼蓝染色：
推荐向900μL细胞悬液中加入100μL台盼蓝染色液轻轻混匀，终浓度为0.04% w/v。根据样品和仪器，终浓度范围从0.04%到0.2%（w/v）。室温染色3min(染色时间可适当延长，但不宜超过10min)。
3. 细胞计数：
吸取少量经过染色后的细胞，用血细胞计数板计数，镜下可观察到死细胞着蓝色并膨大无光泽，活细胞不着色，维持正常形态有光泽。
通常如果要比较精确地进行定量，每个细胞样品至少需要数500个细胞，数出蓝色细胞和细胞总数。细胞存活率计算公式如下：
细胞存活率＝(细胞总数－蓝色细胞数)/细胞总数×100%。
注意事项
1. 台盼蓝对人体有轻微毒性，请注意小心防护。
2. 注意凋亡小体偶尔也有台盼蓝拒染现象。
3. 为了您的安全和健康，请穿实验服并戴一次性手套操作。
结果展示
图 1
图 1. 台盼蓝染色液对 K562细胞存活率测试效果图。细胞1000rpm离心5分钟收集K562细胞，弃上清，用适量细胞重悬液重悬后，分别750μL细胞悬液加入250μL台盼蓝染色液（4×）或 900μL细胞悬液加入100μL台盼蓝染色液（10×）染色3min。台盼蓝染为蓝色的即为死亡细胞。
The Trypan Blue Dyeing Kit is designed for live/dead cell counting and viability assessment in cell culture. Trypan Blue is a vital dye that selectively stains cells with compromised membrane integrity (dead cells) blue, while live cells with intact membranes remain unstained. Based on this principle, dead and live cells can be distinguished microscopically, allowing accurate calculation of cell viability.
Protocol
1. Cell Collection
If collecting adherent cells, digest the cells with trypsin-EDTA solution, then harvest them. For suspension cells, cells can be collected directly. Centrifuge the collected cells at 1000 rpm for 5 minutes, discard the supernatant, and resuspend the cell pellet in 1 mL of cell suspension buffer.
2. Trypan Blue Staining
It is recommended to add 100 μL of Trypan Blue stock solution (0.4%) to 900 μL of the cell suspension and mix gently. The final concentration will be 0.04% (w/v). Depending on the sample and instrument, the final concentration may range from 0.04% to 0.2% (w/v). Stain at room temperature for 3 minutes (staining time can be appropriately extended, but should not exceed 10 minutes).
3. Cell Counting
Transfer a small amount of the stained cell suspension to a hemocytometer for counting. Under the microscope, dead cells appear blue, swollen, and dull, while live cells remain unstained and exhibit normal morphology with a glossy appearance. For accurate quantification, at least 500 cells per sample should be counted. Count both the blue-stained (dead) cells and the total number of cells. Cell viability is calculated using the following formula:
Cell Viability (%) = (Total number of cells – Number of blue cells) / Total number of cells × 100%
Notes
1. Trypan Blue is slightly toxic to humans. Please take proper precautions to avoid exposure.
2. Note that apoptotic bodies may occasionally exhibit Trypan Blue exclusion as well.
3. For your safety and health, please wear a lab coat and disposable gloves during handling.
Results Presentation
Figure 1
Figure 1. Viability test of K562 cells using Trypan Blue staining. Cells were collected by centrifugation (1000 rpm, 5 min), resuspended in cell suspension buffer. Stain for 3 minutes using either 750 μL cell suspension + 250 μL Trypan Blue (4×) or 900 μL cell suspension + 100 μL Trypan Blue (10×). Blue-stained cells were counted as dead cells.
T1511146 | Component | 100T | 500T | Storage |
T1511146A | 台盼蓝染色液 | 10 mL | 50 mL | 2-8℃ |
T1511146B | 细胞重悬液 | 100 mL | 250 mL*2 | 2-8℃ |
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 ZJ26F1039821 |
分析证书 | T1511146 |
| ZJ26F0636853 |
分析证书 | T1511146 |
| ZJ26F0636852 |
分析证书 | T1511146 |
| ZJ26F0636424 |
分析证书 | T1511146 |
| ZJ26F0636425 |
分析证书 | T1511146 |
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