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BioReagent,生物染色剂,用于显微镜 级 ,适用于对基线干扰要求严格的色谱和分析工作流程。
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在色谱分析、有机合成和交叉偶联反应领域已被 53 篇同行评审文献引用。
台盼蓝 (Trypan blue) 又称锥蓝或锥虫蓝等,分子式为 C₃₄H₂₄N₆Na₄O₁₄S₄,分子量为 960.81,CAS Number 72-57-1,是一种阴离子型染料,Trypan blue 不能透过活细胞正常完整的细胞膜,故活细胞不着色,即产生所谓的拒染现象,而死亡细胞的细胞膜通透性增加,可使染料通过细胞膜进入细胞内,使死细胞着色呈蓝色,是最常用的检测细胞活率的方法。通常认为细胞膜完整性丧失,即可为细胞已经死亡,这与中性红作用相反,因此借助台盼蓝染色可以非常简便、快速地区分活细胞和死细胞,台盼蓝是组织和细胞培养中最常用的死细胞鉴定染色方法之一。
Trypan Blue Solution (0.4%) 常用于检测细胞膜的完整性以及细胞是否存活,活细胞不会被染成蓝色,而死细胞会被染成淡蓝色;台盼蓝可被巨噬细胞吞噬,故可用于巨噬细胞的活体染色剂;台盼蓝染色后通过显微镜下直接计数或显微镜下拍照后计数,就可以对细胞存活率进行比较精确的定量,0.4%为最常用的浓度。该试剂仅用于科研领域,不适用于临床诊断或其他用途。
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 1000~2000 g 离心 1 min,弃上清,用 1 mL PBS 重新悬浮细胞沉淀。
2. 台盼蓝染色:吸取 100 μL 细胞悬液到 1.5 mL 或 0.5 mL 离心管内,加入100 μL Trypan Blue Solution (0.4%) 轻轻混匀,静置染色 3 min (染色时间可适当延长,但不宜超过 10 min)。
3. 细胞计数:吸取经过染色后的细胞,用血细胞计数板计数。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。
4. 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:
1. Trypan Blue Solution (0.4%) 对人体有轻微刺激性,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。
免责声明:本公司将不为任何不正常使用此产品时所发生的意外负责。
Trypan blue, also known as Trypan blue stain, has the molecular formula C₃₄H₂₄N₆Na₄O₁₄S₄, a molecular weight of 960.81 and CAS No. 72-57-1. It is an anionic dye. Trypan blue cannot penetrate the intact cell membrane of viable cells, so living cells remain unstained, which is known as the dye exclusion effect. In contrast, the cell membrane permeability of dead cells increases, allowing the dye to enter the cells and stain dead cells blue. It is the most commonly used method for cell viability detection. Generally speaking, the loss of cell membrane integrity indicates cell death, which is opposite to the staining principle of neutral red. Therefore, trypan blue staining enables simple and rapid differentiation between live and dead cells, and it is one of the most widely used staining methods for dead cell identification in tissue and cell culture.
Trypan Blue Solution (0.4%) is commonly used to detect cell membrane integrity and cell viability. Live cells are not stained blue, while dead cells turn pale blue. Since trypan blue can be phagocytosed by macrophages, it can serve as a vital stain for macrophages. After trypan blue staining, cell viability can be accurately quantified via direct microscopic counting or counting based on microscopic images, with 0.4% being the most frequently used working concentration. This reagent is only for scientific research purposes and is not applicable to clinical diagnosis or other uses.
Usage Method:
1. Cell collection
For adherent cells, digest cells with trypsin-EDTA solution before collection; for suspension cells, collect directly. Centrifuge the harvested cells at 1000~2000 g for 1 minute, discard the supernatant, and resuspend the cell pellet in 1 mL PBS.
2. Trypan blue staining
Transfer 100 μL cell suspension into a 1.5 mL or 0.5 mL centrifuge tube, add 100 μL Trypan Blue Solution (0.4%) and mix gently. Let stand for 3 minutes for staining. The staining time can be properly extended, but shall not exceed 10 minutes.
3. Cell counting
Take the stained cell mixture and count cells with a hemocytometer. For accurate quantification, count no less than 500 cells per sample, and record the number of blue-stained cells and total cells.
4. Cell viability calculation formula
Cell Viability = (Total Cell Number − Blue Cell Number) / Total Cell Number × 100%
Precautions:
1. Trypan Blue Solution (0.4%) is slightly irritating to the human body. Please take proper protective measures during operation.
2. Note that apoptotic bodies may occasionally show trypan blue dye exclusion.
3. For your personal safety and health, please wear lab coats and disposable gloves when handling.
Disclaimer: Our company shall not be liable for any accidents caused by improper use of this product.
H350: 可能导致癌症
P280: 戴防护手套/穿防护服/戴防护眼罩/戴防护面具。
P405: 密闭存放
P501: 将内容物/容器处理到。。。
P203: 使用前,获取、阅读并遵守所有安全说明。
P318: 如果暴露或担心,请就医。
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
| 分析证书 | T1523203 | |
| 分析证书 | T1523203 |
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