Search
Search
View additional product information for Vybrant™ DiI Cell-Labeling Solution - FAQs (V22885)
11 product FAQs found
要考虑的因素有示踪对象的大小、给样方式（注射，直接上样到组织等），示踪对象是否需要固定。以下链接详细介绍了我们提供的各类神经元示踪剂的详细信息以及选择方法：
•神经元示踪（https://www.thermofisher.com/us/en/home/life-science/cell-analysis/cell-tracing-tracking-and-morphology/neuronal-tracing.html）
•选择示踪剂（https://www.thermofisher.com/us/en/home/references/molecular-probes-the-handbook/fluorescent-tracers-of-cell-morphology-and-fluid-flow/choosing-a-tracer.html）
•成像分析（http://assets.thermofisher.com/TFS-Assets/BID/Reference-Materials/bioprobes-50-journal.pdf）
这是不推荐的。这些染料与DNA和RNA的结合会影响核酸的正常功能，扰乱转录和增殖。诸如CellTracker染料或Qtracker试剂在不严重扰乱细胞正常活动的条件下对其进行追踪。如果您仍需要使用核酸染料进行标记且细胞是哺乳动物和非血液来源的话，CellLight 细胞核试剂可通过瞬时转染进入细胞，在核表达蛋白上表达GFP或RFP长达数天而不影响其功能。
请浏览这里（https://www.thermofisher.com/us/en/home/life-science/cell-analysis/cell-tracing-tracking-and-morphology/cell-tracking.html）以帮助您选择适合您的应用的产品。首先确定追踪细胞的时间，然后考虑染料结合机制。钙黄绿素染料标记均匀且对短期细胞迁移示踪效果极佳，但也会被某些类型细胞迅速外排。亲脂性的花青素染料，如DiI，DIO，和类似的染料能标记细胞膜而不破坏其功能，并且能持续更长时间，但如果发生膜融合则可能会染上其他细胞。此外，它们还会在透化过程中丢失。CellTracker染料更有利于长期标记，其带有温和的氯甲基反应基团使之能够与细胞组分共价结合。CFDA SE也能共价地结合于细胞组分。在所有列出的试剂中，细胞内保留与否取决于细胞分裂的速率和细胞的固有特性（主动外排，膜和蛋白质的周转率等）。其中共价结合试剂比非共价结合的试剂展现出更长的保留时间。
Qtracker试剂是最持久并且荧光强度最高的细胞示踪染料，它通过内吞作用被细胞摄入。在许多样品中它们产生的信号可以持续检测长达数周，而且信号足够强，即使在固定和通透甚至加热和石蜡处理过程中，仍然能够维持较好的荧光信号。
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。
This is expected. DiD (which is far-red fluorescent) is never as uniform as DiI (which is orange fluorescent). If uniformity is desired, try increasing the label time and concentration, but it still isn't likely to be as uniform as DiI. CellMask Deep Red Plasma Membrane stain is much more uniform and is about the same wavelength as DiD. However, if you intend to do cell tracking over days, CellMask stain has not been tried for that application.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
A typical method is to label one cell line with orange fluorescent DiI C18 and the other cell line with green fluorescent DiO C18. These orange and green lipophilic cyanine dyes will stain the membranes of cells. Cells that fuse will then have both dyes, yielding a yellow color (when images are overlaid or cells are imaged in a dual-bandpass filter). These live cells can then be labeled with Hoechst 33342 (a cell-permeant blue DNA stain comparable in wavelength to DAPI), but only as an endpoint just before imaging (since DNA stains can interrupt DNA function).
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
Lipophilic cyanine dyes, such as DiI (Cat. No. D282), DiO (Cat. No. D275), DiD (Cat. No. D7757) or DiR (Cat. No. D12731), are commonly used. The longer the alkyl chain on the dye, the better the retention in lipophilic environments.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
Factors to consider are size of tracer, method of delivery (injection, direct application to tissue, etc.), and if the tracer needs to be fixable. Here are some links to details about the various classes of neuronal tracers we offer and how to choose between them:
Neuronal Tracing (https://www.thermofisher.com/us/en/home/life-science/cell-analysis/cell-tracing-tracking-and-morphology/neuronal-tracing.html)
Choosing a Tracer (https://www.thermofisher.com/us/en/home/references/molecular-probes-the-handbook/fluorescent-tracers-of-cell-morphology-and-fluid-flow/choosing-a-tracer.html)
Imaging Analysis (http://assets.thermofisher.com/TFS-Assets/BID/Reference-Materials/bioprobes-50-journal.pdf)
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
This is not recommended. When these stains bind to DNA and RNA, they may affect the normal function of the nucleic acids, disrupting transcription, as well as replication. Other reagents, such as CellTracker dyes or Qtracker reagents are more optimized for tracking without disrupting normal activity. If a nuclear label is still desired, though, and the cells are mammalian and non-hematopoietic, CellLight nuclear reagents can transiently transfect cells to express GFP or RFP on a nuclear-expressing protein for up to several days without affecting function.
Find additional tips, troubleshooting help, and resources within our Cell Tracing and Tracking Support Center.
Please see this Web link (http://www.thermofisher.com/us/en/home/life-science/cell-analysis/cell-tracing-tracking-and-morphology/cell-tracking.html) to help you choose the right option for your application. Start by planning how long you want to track your cells, then consider the mechanism of binding. Calcein dyes are very uniform in label and are good for short-term cell migration, but may be rapidly effluxed from some cell types. Lipophilic cyanine dyes, such as DiI, DiO, and similar dyes label cell membranes, don’t disrupt function, and can last longer, but have the potential to cross to other cells if membranes fuse. They are also lost upon permeabilization. CellTracker dyes are better for longer-term labeling, as they possess a mildly reactive chloromethyl moiety that allows covalent binding to cellular components. CFDA SE also covalently binds to cellular components. With all the reagents, their retention within cells is dependent upon the rate of cell division and the inherent properties of the cell (active efflux, membrane and protein turnover rates, etc.) and reagents that allow for covalent attachment exhibit longer retention than those that do not.
The longest-lasting and brightest options are the Qtracker reagents, which are taken up through endocytosis. These are so bright individual quantum dots can be detected, and are also robust enough to survive not only fixation and permeabilization, but even the heat and solvents used in paraffin processing.
Find additional tips, troubleshooting help, and resources within our Cell Tracing and Tracking Support Center.
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.