CytoMBrite™ cell membrane orange red fluorescent probe(Dil)

CytoMBriteTM细胞膜橙红色荧光探针（DiI）是一种亲脂性碳菁类染料，能够有效、稳定地标记质膜及细胞内膜结构。它具有细胞毒性低且不会在细胞间转移的特性，常被作为细胞融合、粘附、迁移的示踪分子，与PKH类染料相比，CytoMBrite染料使用方便、着色均匀。DiI（橙色荧光）、DiO（绿色荧光）、DiD（红色荧光）和其它细胞膜荧光染料如DiR（近红外荧光）、NIR680（远红外染料）配合使用，为多色成像和流式细胞分析提供了有效的工具。 CytoMBrite系列染料也可以用于甲醛固定后细胞的染色，同时兼容在染色后的甲醛固定步骤。此系列染料不适用于细菌或酵母。 以每次使用100 μL染色工作液计算，200 μL原液可以用400次。 产品参数： Ex/Em(MeOH) = 549/565 nm 分子式：C59H97ClN2O4 分子量：933.9 注意事项： 1. 使用前请将产品顺时离心至管底，再进行后续试验。 2. 荧光染料均存在淬灭问题，请尽量注意避光，以减缓荧光淬灭。 3. 为了您的安全和健康，请穿实验服并戴一次性手套操作。 应用范围： 细胞膜染料 产品特点 ： 毒性低 ：具有细胞毒性低且不会在细胞间转移； 稳定性好 ：荧光亮度强、着色均匀且适合细胞示踪，可以在细胞内很好 的保留； 兼容性强 ：甲醛固定前后均可进行染色。 光谱图： 自备材料： 1. 耗材 (1)离心管 (2)盖玻片 2. 试剂 (1)无血清培养基或 HBSS 或 PBS (2)培养基（预温） 3. 仪器 荧光显微镜或流式细胞仪 操作步骤： 工作液使用体积建议根据不同细胞系和实验体系来优化。建议从推荐量的 1010 倍范围内开始最优条件的摸索。 1. 悬浮细胞染色 (1)加入适当体积的培养基重悬细胞，使其密度为 1×106/mL，再按 1：200 的比例加入染色原液。 (2)37℃孵育细胞 2~20 min，不同的细胞最佳培养时间不同。可以 20min 作为起始孵育时间，之后优化体系。 (3)1000~1500 rpm 离心 5 min。倾倒上清液，再次缓慢加入 37℃预热的培养基重悬细胞。重复两次。 2. 贴壁细胞染色 (1)配制染色工作液：每 1 mL 培养基中加入 5 μL 的染色原液，涡旋混匀。 (2)将贴壁细胞培养于无菌盖玻片上，培养结束后移出盖玻片，吸走过量培养液，但表面要保持湿润。 (3)在盖玻片的一角加入 100 μL 的染色工作液，轻轻晃动使染料均匀覆盖所有细胞。 (4)37℃孵育细胞 2~20 min，不同的细胞最佳培养时间不同。可以 20min 作为起始孵育时间，之后优化体系以得到均一的标记效果。 (5)吸干染料工作液，用培养液洗盖玻片 2~3 次，每次用预温的培养基覆盖所有细胞，孵育 5~10 min，然后吸干培养基。但要使表面保持湿润。 3. 结果检测 样品可在培养基中进行检测，可通过荧光显微镜成像或流式细胞仪分析。 注： 黄色光激发，荧光显微镜滤光片可以选择 Cy3 滤光片；流式细胞仪选择 YL1 通道。 Product introduction： Cytombritetm cell membrane orange red fluorescent probe (DII) is a lipophilic carbocyanine dye, which can effectively and stably label the plasma membrane and intracellular membrane structures. It has the characteristics of low cytotoxicity and will not transfer between cells. It is often used as a tracer molecule for cell fusion, adhesion and migration. Compared with PKH dyes, cytombrite dyes are convenient to use and evenly colored. DiI (orange fluorescence), DIO (green fluorescence), did (red fluorescence) and other cell membrane fluorescent dyes, such as dir (near-infrared fluorescence) and nir680 (far-infrared dye), provide effective tools for multicolor imaging and flow cytometry analysis. Cytombrite series dyes can also be used for the staining of cells after formaldehyde fixation, and are compatible with the formaldehyde fixation step after staining. This series of dyes is not suitable for bacteria or yeast. At 100 per use μ L dyeing working solution calculation, 200 μ L stock solution can be used for 400 times. Product parameters： Ex/em (MeOH) = 549/565 nm molecular formula: c59h97cln2o4 molecular weight: 933.9 Matters needing attention： 1. before use, centrifuge the product to the bottom of the tube in time, and then conduct subsequent tests. 2. fluorescent dyes have quenching problems. Please try to avoid light to slow down fluorescence quenching. 3. for your safety and health, please wear experimental clothes and disposable gloves. Scope of application： Cell membrane dye Product Features： Low toxicity: It has low cytotoxicity and will not transfer between cells; Good stability: strong fluorescence brightness, uniform coloring, and suitable for cell tracking, which can work well inside cells The reservation; Strong compatibility: Dyeing can be performed before and after formaldehyde fixation. Spectral diagram: Bring your own materials： 1. Consumables (1) Centrifuge tube (2) cover glass 2. Reagents (1) Serum free medium or HBSS or PBS (2) medium (pre warmed) 3. Instruments Fluorescence microscope or flow cytometer Operating steps： It is recommended to optimize the volume of working fluid according to different cell lines and experimental systems. Suggest starting to explore the optimal conditions within the range of 1010 times the recommended quantity. 1. Suspension cell staining (1) Add an appropriate volume of culture medium to resuspend the cells to a density of 1 × 106/mL, and then add the staining solution in a ratio of 1:200. (2) Incubate cells at 37 ℃ for 2-20 minutes, and the optimal culture time varies for different cells. A starting incubation time of 20 minutes can be used to optimize the system afterwards. (3) Centrifuge at 1000~1500 rpm for 5 minutes. Pour the supernatant and slowly add the culture medium preheated at 37 ℃ again to resuspend the cells. Repeat twice. 2. Staining of adherent cells (1) Preparation of staining solution: Add 5 μ L of staining solution to every 1 mL of culture medium, vortex and mix well. (2) Cultivate the adherent cells on sterile cover glass, remove the cover glass after cultivation, absorb excess culture medium, but keep the surface moist. (3) Add 100 μ L of staining solution to one corner of the cover glass and gently shake to evenly cover all cells with the dye. (4) Incubate cells at 37 ℃ for 2-20 minutes, and the optimal culture time varies for different cells. 20 minutes can be used as the starting incubation time, and then the system can be optimized to achieve uniform labeling effect. (5) Absorb the dye working solution, wash the cover glass with culture medium 2-3 times, cover all cells with preheated culture medium each time, incubate for 5-10 minutes, and then absorb the culture medium. But keep the surface moist. 3. Result detection The sample can be detected in the culture medium and analyzed by fluorescence microscopy imaging or flow cytometry. Note: Yellow light excitation, Cy3 filter can be selected as the fluorescence microscope filter; Select YL1 channel for flow cytometry.

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