DiO Plus（细胞膜绿色荧光探针升级款）

未知

Dio plus (cell membrane green fluorescent probe upgrade)

D598343-5mg

5mg

2-8°C储存,避光

冰袋运输

DiO Plus效能等同于Neuro-DiO。DiO Plus是一种代替膜绿色荧光染料DiO的新型染料，DiO由于其溶解度低、易形成聚合物和横向扩散速度缓慢等缺点，很难用于神经元和细胞悬液的研究。DiO Plus和DiO具有几乎相同的吸收光谱和发射光谱，但前者具有更好的膜溶解性，并且不形成非荧光性的聚集体，通常这也会减慢染料在膜上的扩散速率。DiO Plus染色后可进行多聚甲醛（不可使用甲醇等其他试剂）的固定，但不建议在染色后进行透化的过程。此外，在固定透化（室温下用0.1% TritonX-100 透化）后，也可以很好地进行质膜染色。以每次使用100 μL染色工作液，染色工作液浓度3 μM计算，10 mg配置为工作液大概可以用15346次。 产品参数： Ex/Em (MeOH) = 484/501 nm 应用范围： 细胞膜荧光染料；神经元顺行和逆行示踪；细胞长期示踪 使用方法： 1. 染色液制备 （1）配置 DMSO 或 EtOH 储存液：储存液用 DMSO 或 EtOH 配置浓度 1~5 mM。 注：未使用的储存液分装储存在-20℃，避免反复冻融。 （2）工作液制备：用合适的缓冲液（如：无血清培养基，HBSS 或 PBS）稀释储存液，配制浓度为 1~5 μM 的工作液。 注：工作液最终浓度建议根据不同细胞系和实验体系来优化建议从推荐浓度的 10 倍范围内开始最优浓度的摸索。 2. 悬浮细胞染色 （1）加入适当体积的染色工作液重悬细胞，使其密度为 1×106/mL。 （2）37℃孵育细胞 20 min，不同的细胞最佳培养时间不同。可以 20 min 作为起始孵育时间，之后优化体系以得到均一的标记结果。 （3）孵育结束，1000~1500 rpm 离心 5 min。倾倒上清液，再次缓慢加入 37℃预热的生长培养液重悬细胞。 （4）重复步骤（3）两次以上。 3. 贴壁细胞的染色 （1）将贴壁细胞培养于无菌盖玻片上。 （2）从培养基中移走盖玻片，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 的染料工作液，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 20 min 作为起始孵育时间，之后优化体系以得到均一的标记结果。 （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。 4. 结果检测 样品可在培养基中进行检测，可以通过荧光显微镜成像或流式细胞仪分析。 注意事项： 1. 使用前请将产品瞬时离心至管底，再进行后续实验。 2. DiO Plus 染色固定的细胞或组织样品时，通常使用配制在 PBS 中的 4%多聚甲醛进行固定，使用其它不适当的固定液会导致荧光背景较高。 3. 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。 4. 为了您的安全和健康，请穿实验服并戴一次性手套操作。 The efficiency of DiO Plus is equivalent to Neuro-DiO. DiO Plus is a new type of dye that replaces the membrane green fluorescent dye DiO.Due to its low solubility, easy formation of polymers and slow lateral diffusion, DiO is difficult to be used in the study of neurons and cell suspensions. DiO Plus and DiO have almost the same absorption and emission spectra, but the former has better membrane solubility and does not form non-fluorescent aggregates, which usually slows down the diffusion rate of dyes on the membrane. The fixation of paraformaldehyde ( other reagents such as methanol cannot be used ) can be carried out after dyeing with DiO Plus, but the process of dialysis after dyeing is not recommended. In addition, plasma membrane staining can also be performed well after fixed permeabilization ( permeabilization with 0.1 % TritonX-100 at room temperature ). Based on the use of 100 μL dyeing working fluid at a time, the concentration of the dyeing working fluid is 3 μM, and 10 mg is configured as the working fluid, which can be used about 15,346 times. Product parameters: Ex/Em (MeOH) = 484/501 nm Scope of application： Cell membrane fluorescent dye ; anterograde and retrograde tracing of neurons ; long-term cell tracing Usage: 1. Preparation of staining solution (1) Configure DMSO or EtOH storage solution: Use DMSO or EtOH to prepare the storage solution with a concentration of 1-5 mM. Note: Unused storage liquids should be packaged and stored at -20 ℃ to avoid repeated freeze-thaw cycles. (2) Preparation of working solution: Dilute the storage solution with a suitable buffer (such as serum-free medium, HBSS or PBS) and prepare a concentration of 1-5 μ M's working fluid. Note: It is recommended to optimize the final concentration of the working solution based on different cell lines and experimental systems. It is recommended to start exploring the optimal concentration within a range of 10 times the recommended concentration. 2. Suspension cell staining (1) Add an appropriate volume of staining solution and resuspend the cells to a density of 1 × 106/mL. (2) Incubate cells at 37 ℃ for 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 obtain uniform labeling results. (3) After incubation, centrifuge at 1000-1500 rpm for 5 minutes. Pour the supernatant and slowly add the growth culture medium preheated at 37 ℃ again to resuspend the cells. (4) Repeat step (3) more than twice. 3. Staining of adherent cells (1) Cultivate adherent cells on sterile cover slips. (2) Remove the cover glass from the culture medium, absorb excess culture medium, but keep the surface moist. (3) Add 100 to one corner of the cover glass μ Gently shake the dye working solution of L 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 obtain uniform labeling results. (5) Dry 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 dry the culture medium. But keep the surface moist. 4. Result detection The sample can be detected in the culture medium and analyzed by fluorescence microscopy imaging or flow cytometry. Notes: 1. Before use, please centrifuge the product instantaneously to the bottom of the tube before conducting subsequent experiments. 2.When staining fixed cell or tissue samples with DiO Plus, 4% paraformaldehyde prepared in PBS is usually used for fixation. The use of other inappropriate fixation solutions can result in a high fluorescence background. 3. Fluorescent dyes all have quenching problems, please try to avoid light as much as possible to slow down fluorescence quenching. 4.For your safety and health, please wear laboratory clothes and disposable gloves when operating.

C67H105ClN2O5S

1086.00 g/mol

可溶于乙醇、己烷、DMSO或植物油