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View additional product information for Dextran, Fluorescein, 10,000 MW, Anionic, Lysine Fixable (Fluoro-Emerald) - FAQs (D1820)
10 product FAQs found
如果您选择的示踪剂是亲脂性染料并且用甲醇固定，脂质会随着甲醇流失。如果您使用甲醇固定，可以选择能够共价结合神经元内的蛋白质的示踪剂。
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。
DiI是一种亲脂性染料，多数保留在细胞脂质内。当细胞经去垢剂通透或是用乙醇固定时，会去除掉脂质和染料。如果需要通透，可以使用CM-DiI标记，因为CM-DiI会共价结合膜里面的蛋白；部分信号会随着固定/通透丢失，但残留的信号仍足以被检测出。
固体和结晶形式的DiI以及其他相关染料（货号D282，D3911，D7757，和D12731）有时与一个特定的神经元接触，通过膜侧向扩散染上整个细胞。此外，我们的NeuroTrace组织标记糊剂也可挑到针上，置于特定的神经元上。 有关我们的神经元细胞标记方法的比较结果请参阅下表。 产品：标记方式：标记强度：特征 神经元特异性抗体：针对神经细胞表达蛋白的一抗：与蛋白质表达量成比例：唯一的神经元特异性标记方式。 亲脂性神经元示踪剂：疏水性染料掺入到细胞中的脂质：由于细胞中含有大量的脂类，这种标记方式能提供最强的标记：能够跟踪整个样品的神经元。 荧光标记的右旋糖苷：通常右旋糖苷采用显微注射：所有的细胞质可能被标记，标记密度可能会更高：能够标记单个神经元，从而避免了荧光背景干扰。 膜电位指示剂：染料通过水性缓冲液上样到活细胞里：取决于周围电场引起的结构变化，或者去极化引起的染料流入：膜电位的变化在各种生理进程中发挥核心作用，包括神经脉冲波传播，肌肉收缩以及细胞信号传导过程。
详细信息请查阅此网页（https://www.thermofisher.com/us/en/home/life-science/cell-analysis/cell-tracing-tracking-and-morphology/neuronal-tracing.html）。
If the tracer you chose is a lipophilic dye and fix with methanol, the lipids are lost with the methanol. If you have to use methanol fixation then choose a tracer that will covalently bind to proteins in the neurons.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
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.
DiI is a lipophilic dye that resides mostly in lipids in the cell, when cells are permeabilized with detergent or fixed using alcohol this strips away the lipid and the dye. If permeabilization is required CM-DiI can be used because this binds covalently to proteins in the membrane; some signal is lost upon fixation/permeabilization, but enough signal should be retained to make detection possible.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
The solid and crystalline forms of DiI and other related dyes (Cat. Nos. D282, D3911, D7757, and D12731) are sometimes placed in contact with a specific neuron where it will travel down the cell by lateral diffusion via the membrane. Alternatively, our NeuroTrace Tissue Labeling Paste can be scooped onto a needle and placed onto particular neurons.
Please see the information below for a comparison of our neuronal cell labeling methods:
Product:Method of labeling: Labeling intensity: Features
Neuron-specific antibodies: Primary antibodies directed to proteins expressed in neuronal cells: Proportional to the amount of protein expressed: Provides the only neuronal specific labeling method
Lipophilic neuronal ytracers: Hydrophobic dyes are incorporated into lipids in the cell: This labeling method provides the most intense labeling becuase of the abundant amount of lipids: Allows tracing of neurons throughout the sample
Membrane potential indicators: Dyes are loaded into live cells in aqueous buffers: Depends on either changes in structures due to the electrical field they are in, or dye influx due to depolarization: Changes in membrane potential play a central role in physiological processes, including nerve-impulse propagation, muscle contraction, and cell signaling
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
Please check out this web page (https://www.thermofisher.com/us/en/home/life-science/cell-analysis/cell-tracing-tracking-and-morphology/neuronal-tracing.html) for details.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.