BackDrop™ Background Suppressor ReadyProbes™ Reagent - FAQs

View additional product information for BackDrop™ Background Suppressor ReadyProbes™ Reagent - FAQs (R37603)

5 product FAQs found

How do I quench any fluorescence outside of my live cells and leave my intracellular staining intact?

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.

The BackDrop background suppressor solutions are supposed to reduce background fluorescence. Does that include sample autofluorescence?

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.

Can I use the ReadyProbes reagents for flow cytometry?

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.

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.

I am not seeing a strong degree of change in my ion indicator dye in my positive versus negative controls. How can I correct this?

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.