Neurobasal™ Medium, minus phenol red - FAQs

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我知道有些培养基进行荧光成像的效果不佳，该如何选择培养基呢?

大多数培养基都含有酚红，会淬灭可见光波长范围的荧光染料。大多数培养基还含有自发荧光成分——如核黄素——会降低信噪比。我们提供的FluoroBrite DMEM和基于HEPES的活细胞成像溶液已针对荧光成像应用进行了优化。此外，我们还提供多种不含酚红的培养基。但是，如果这些产品都不是您的试验的合理选择，为此我们还提供BackDrop 背景抑制剂 ReadyProbes 试剂，将其加入可淬灭培养基的自发荧光。

使用荧光染料标记活细胞时，我是否需要考虑培养基中酚红的影响？

一些类型的细胞会积聚酚红，阻碍使用荧光探针。酚红会淬灭可见光波长范围内的染料。此外，尽管酚红本身并不发荧光，但其中的各种杂质可能会发出荧光。我们有许多不含酚红的培养基可供选择。我们的活细胞成像溶液（基于HEPES）和FluoroBrite DMEM已经过优化，不含酚红且无自发荧光。

为何不含酚红的Neurobasal培养基的颜色为黄色？

黄色是由维生素成份所带来的。

在培养神经细胞的过程中，我可在Neurobasal培养基中加入哪些添加剂？

做为一般性的参考方案，我们推荐您使用Gibco B-27添加剂来培养神经干细胞，海马及其他类型的CNS神经元。

-对于来自PNS和CNS中的神经母细胞瘤或有丝分裂后的神经元，可添加Gibco N-2添加剂。
-对于原代胶质细胞（星形胶质细胞）或具有星形胶质细胞表型的肿瘤细胞系（星形胶质细胞和胶质细胞瘤），或少突胶质细胞，可使用Gibco G-5添加剂。

Can Neurobasal Medium alone or Neurobasal Medium supplemented with B-27 supplement be frozen to extend product shelf life?

No, it is not recommended to freeze media due to the potential of precipitates forming upon thaw. Inorganic salts and amino acids in the formulation may come out of solution when exposed to temperature fluctuations. These precipitates will not go back into solution easily once formed.

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

What supplements should I use with Neurobasal or Neurobasal-A medium?

Neurobasal and Neurobasal-A media (for postnatal and adult neurons) allow for long-term maintenance of neuronal cells without the need for an astrocyte feeder layer. These media should be supplemented with either serum or a serum-free supplement, plus 0.5mM L-glutamine. B-27 supplement is a serum-free supplement that comes as a 50X concentrate in a 10ml volume. This is enough supplement for 500ml of media. Fetal, postnatal, and adult neural cultures can be grown in the appropriate Neurobasal medium supplemented with B-27 supplement .

We also have two other supplements. One is called G-5 and is for growth and expression of glial cells (normal and tumor) of astrocytic phenotype. This comes in a 1ml size, at a 100X concentration. The other supplement is called N-2 and is for growth and expression of post-mitotic neurons and tumor cells of neuronal phenotype. This comes in a 5ml size, at a 100X concentration.

For more information on Neurobasal media, search "Neurobasal" from our website home page.

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

Does the removal of phenol red from the Neurobasal Medium affect the culturing of neurons?

No, removal of phenol red from the Neurobasal Medium does not affect the culturing of neurons. Removal of phenol red allows culturing of neurons for studies involving certain receptors and downstream purification.

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

When should I use the Neurobasal Medium, minus phenol red?

We recommend using Neurobasal Medium, minus phenol red for studies involving certain receptors, downstream protein purification or other processes where the presence of phenol red is undesirable.

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

I understand that some media are worse than others for fluorescence imaging. How do I choose?

Most media contain phenol red, which can quench fluorescent dyes in the visible wavelengths. Most media also contain autofluorescent components, such as riboflavin, which can reduce signal-to-background. We offer FluoroBrite DMEM and HEPES-based Live Cell Imaging Solution, which have been optimized for fluorescent imaging. We also offer a number of media without phenol red. But if none of these are reasonable options for your experiment, then we also offer BackDrop Background Suppressor ReadyProbes Reagent, which can be added to quench media autofluorescence.

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

Should I be concerned about phenol red in my media when labeling my live cells with fluorescent dyes?

Some cell types accumulate phenol red, and this can pose a problem in the use of many fluorescent probes. Phenol red can quench visible-wavelength dyes and, although phenol red is non-fluorescent, various impurities may be fluorescent. We have many phenol red-free media to choose from. Our Live Cell Imaging Solution (HEPES-based) and our FluoroBrite DMEM have been optimized to be phenol red-free as well as to be non-autofluorescent.

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

Why is the color of Neurobasal Medium without Phenol red yellow?

The yellow color comes from vitamin components.

Which supplement can I add to Neurobasal Medium to culture my neural cells?

As a general guideline, we recommend using Gibco B-27 Plus Supplement to culture neural stem cells, hippocampal, and other CNS neurons.

- For neuroblastomas, or post-mitotic neurons from both PNS and CNS, Gibco N-2 Supplement can be used.
- For primary glial cells (astrocytes) or tumor cell lines of astrocytic phenotype (astrocytes and gliomas), or oligodendrocytes, Gibco G-5 Supplement can be used.