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DiOC6(3) (3,3'-Dihexyloxacarbocyanine Iodide) - FAQs

DiOC6(3) (3,3'-Dihexyloxacarbocyanine Iodide) - FAQs

查看更多产品信息 DiOC6(3) (3,3'-Dihexyloxacarbocyanine Iodide) - FAQs (D273)

8 个常见问题解答

我用DiI一类的亲脂性花青染料进行细胞染色，但是当我试图继续用抗体标记时信号却丢失了。 这是什么原因所致？

由于这些染料插入脂质膜，任何对膜的破坏都将导致染料流失。这包括用Triton X-100之类的去垢剂或是甲醇之类的有机溶剂通透。通透是胞内抗体标记所必需的，但它会导致染料流失。相反，CFDA SE之类活性染料能够共价连接到细胞组分，因此可以在固定和通透之后更好的保留。

亲脂性示踪剂跨膜转运需要多久？FAST亲脂性染料能快多少？

转运过程相当缓慢：在活体组织中大约6 mm/天，在固定组织中更慢。亲脂性羰花青示踪剂从开始给样处扩散到神经元末端，可能需要数天甚至数周的时间。FAST DIO和DiI类似物（具有不饱和烷基尾）可以提高50％左右的转运速度。

我应该使用哪种亲脂性示踪剂（DiO、DiI、DiD等）？

请选择与您现有的激发光源和发射滤光片组/通道兼容的染料。固体、糊和晶体形式的染料可直接施加到组织中的神经元。如需标记培养的细胞或进行显微注射，可采用溶液或固态形式的亲脂性染料。

我想标记两种细胞群体然后进行细胞融合实验，哪种试剂最适合来成像？

亲脂性花青染料非常适合此类测定，因为它们会掺入到细胞膜中，随着细胞融合之后共用细胞膜，染料也随之共享。例如，一个细胞群体可以用DiI（橙红色）进行标记，而另一细胞群可以用DIO（绿色）进行标记，当细胞融合时合并的颜色为黄色（使用双带通滤光片成像）。

I stained my cells with a lipophilic cyanine dye, like DiI, but the signal was lost when I tried to follow up with antibody labeling. Why?

Since these dyes insert into lipid membranes, any disruption of the membranes leads to loss of the dye. This includes permeabilization with detergents like Triton X-100 or organic solvents like methanol. Permeabilization is necessary for intracellular antibody labeling, leading to loss of the dye. Instead, a reactive dye such as CFDA SE should be used to allow for covalent attachment to cellular components, thus providing for better retention upon fixation and permeabilization.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

How long does it take for lipophlic tracers to transport along the membrane? How much faster are the FAST lipophilic dyes?

The transport is fairly slow, around 6 mm/day in live tissue and slower in fixed tissue, so diffusion of lipophilic carbocyanine tracers from the point of their application to the terminus of a neuron can take several days to weeks The FAST DiO and DiI analogs (which have unsaturated alkyl tails) can improve transport rate by around 50%.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Which form of the lipophilic tracers (DiO, DiI, DiD, etc) should I use?

Select the dye that is compatible with your available excitation source(s) and emission filter set/channels. The solid, paste and crystal forms can be applied directly to neurons in tissues. For labeling cells in culture or microinjection, the lipophilic dyes in solution or solid form can be used.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

I want to label two cell populations and then perform a cell fusion assay. Which reagents are best for imaging this?

Lipophilic cyanine dyes are preferred for this sort of assay, since they insert into cellular membranes and then, upon fusion, are shared by the fused cells as the membranes are shared. For example, one cell population can be labeled with DiI (orange-red) and another cell population can be labeled with DiO (green), and when the cells fuse, the combined color appears yellow (when imaged with a dual-bandpass filter set).

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.