BackDrop™ Background Suppressor for Microplate or Imaging Assays, 50 mL - FAQs

View additional product information for BackDrop™ Background Suppressor for Microplate Assays - FAQs (B10512)

7 product FAQs found

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。

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 the Fluo-4 Calcium Imaging Kit (Cat. No. F10489) components be purchased separately?

Yes. The following components are sold separately:
Component B: PowerLoad Concentrate (Cat. No. P10020)
Component C: BackDrop Background Suppressor for Microplate Assays (Cat. No. B10512)
Component D: Probenecid, Water Soluble (Cat. No. P36400)

Can I use the BackDrop Background Suppressor reagent as the quencher to reach the inner leaflet of the plasma membrane?

We have not tested this. The BackDrop Background Suppressor reagent is cell impermeant, but if the quenching is caused by a FRET interaction and, if the distance between the BackDrop Background Suppressor outside of the cell is within the Ro distance for FRET with the fluorescent dye in the inner leaflet of the membrane, quenching may still occur.

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

Would the BackDrop Background Suppressor reagent quench Alexa Fluor 647 signal? What is the mechanism of quenching?

Yes, the BackDrop Background Suppressor reagent can be used to quench Alexa Fluor 647 signal. We have not determined the exact mechanism for the quenching but it depends upon the dye. For some dyes, it may be a FRET interaction, for others the BackDrop Background Suppressor absorbs the photons emitted by the fluorescent dye (inner filter effect).

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.