BackDrop™ Background Suppressor ReadyProbes™ Reagent - FAQs

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。

首先，确保在适当的波长中激发和检测染料。接下来，尝试优化对照的染料浓度和染色时间。我们的手册有一些指导。如果您使用的是活细胞系统，对于我们的许多产品，我们还建议洗去未反应的染料以减少背景荧光，或通过背景抑制试剂如 BackDrop Suppressor ReadyProbes Reagent抑制背景。请检查您的仪器设置；例如一些读板仪可通过调整增益设置的手段，以获得最佳信噪比。最后，一些染料在离子变化程度检测方面比其他染料好。如果想了解如何选择染料，可发送邮件至techsupport@lifetech.com联系技术支持。

There are a number of reagents used to quench fluorescence on the outside of live cells or in the media (background fluorescence). Our Backdrop Background Suppressor reagents are a great option to quench blue, green, and red fluorescent dyes, various media components and fluorescent proteins. Another commonly used reagent is the cell impermeant colorimetric dye, Trypan blue.

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

No. The BackDrop dyes will quench any fluorescence of a given wavelength in the extracellular media or buffer, but not inside live cells. It will not quench autofluorescence inside live cells unless the dye is engulfed.

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

This is not recommended. The ReadyProbes reagents were developed for imaging applications whereas the Ready Flow reagents were optimized for flow cytometry.

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

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.

First, make sure you are exciting and detecting the dye in the appropriate wavelengths. Next, try optimizing the dye concentration with your controls, as well as the dye staining time. Our manuals have some guidelines. If you are using a live-cell system, we also recommend washing out any unreacted dye for many of our products to reduce background fluorescence, or adding a background suppressor such as BackDrop Suppressor ReadyProbes Reagent. Check your instrument settings; some plate readers, for instance, may have a means of adjusting the gain setting to get the best signal-to-background. Finally, some dyes are better than others for degree of change upon ion detection. Contact Tech Support by sending an email to techsupport@thermofisher.com if you would like to discuss dye options.

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