我想标记细胞膜，但可选的染料有多种，该如何选择？
如果进行活细胞成像，CellVue和CellMask 质膜染色剂染色最均匀，被细胞内吞的速度最慢。如果想固定和通透细胞（如进行抗体标记），这种产品并不是最佳选择。麦胚凝集素（WGA）偶联物也能标记活细胞，或者甲醛固定的细胞。它们可以在随后的去垢剂（如TritonX-100）通透处理后保存下来。但如果细胞已经进行通透处理，WGA也会标记内部结构。因此，如果细胞已通透，则只能使用靶向质膜蛋白的抗体。亲脂性花青染料（如DiI）会标记活细胞的所有膜结构，而不仅仅是细胞质膜。此网页（https://www.thermofisher.com/us/en/home/life-science/cell-analysis/cell-structure/plasma-membrane.html）的内容可帮助您进行选择。
What kind of filter sets can I use with HCS LipidTOX neutral lipid stains?
LipidTOX Green neutral lipid stain can be imaged with filter sets appropriate for Alexa Fluor 488 dye or fluorescein. LipidTOX Red neutral lipid stain is best imaged with filter sets appropriate for Alexa Fluor 594 dye or Texas Red dye. LipidTOX Deep Red neutral lipid stain can imaged with filter sets appropriate for Alexa Fluor 647 dye or Cy5 dye.
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Is the HCS LipidTOX Deep Red Neutral Lipid Stain (Cat. No. H34477) ready to use, or do I need to dilute it in 1X PBS or DMSO?
The HCS LipidTOX Deep Red Neutral Lipid Stain is provided as a solution in DMSO and needs to be diluted before use. The recommended dilution is 1:200 in 1X PBS or other buffer (e.g. HBSS, TBS, etc).
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What buffer should I use with HCS LipidTOX™ Deep Red Neutral Lipid Stain, for cellular imaging (Cat. No. H3447)?
The ideal buffer with HCS LipidTOX Deep Red Neutral Lipid Stain is PBS at pH 7.4.
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I want to label the plasma membrane of my cells, but there are several dyes to choose from. Which one should I use?
For live-cell imaging, the CellVue and CellMask Plasma Membrane Stains are the most uniform and the slowest to be endocytosed. However, they are not the best choice if you wish to fix and permeabilize your cells, such as for antibody labeling. Wheat germ agglutinin (WGA) conjugates are also able to label live cells, or can label already formaldehyde-fixed cells. They can survive subsequent permeabilization with detergents, such as Triton X-100. If cells are already permeabilized, WGA will label internal structures as well. Thus, only an antibody against a plasma membrane protein can be used if cells are already permeabilized. Lipophilic cyanine dyes, such as DiI, will label all cell membranes in live cells, not just plasma membranes, if left on live cells for extended periods. Following page will help you choose (http://www.thermofisher.com/us/en/home/life-science/cell-analysis/cell-structure/plasma-membrane.html).
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
