Stellaris® FISH Probes, Custom Assay with Quasar® 570 Dye

Custom Stellaris® RNA FISH Probe Set labeled with Quasar® 570 dye to detect your RNA of interest.

Product Description

Stellaris RNA FISH is a RNA visualization method that allows simultaneous detection, localization, and quantification of individual RNA molecules at the cellular level in fixed samples using fluorescence microscopy.

A set of Stellaris RNA FISH Probes is comprised of a pool of up to 48 unique probes, each labelled with a fluorophore, that collectively bind along an RNA target transcript to produce a diffraction limited spot.

Before considering designing a Stellaris RNA FISH Customized Probe Set, please also check out our Stellaris RNA FISH DesignReady Probe Sets for a large selection of professionally designed targets.

We also recommend selecting a Stellaris RNA FISH Positive Control (ShipReady) Probe Set RNA  to complement your experiment. 

Stellaris Buffers are the perfect accompaniment to your experiment, allowing for even more robust RNA detection and analysis through reduced background signal. 

Quasar 570 Dye

The Quasar 570 modification incorporates a fluorescent Quasar 570 dye at the terminus of an oligonucleotide. Quasar 570 fluorophore is an indocarbocyanine dye which fluoresces in the yellow-orange region of the visible spectrum. It is used as a direct replacement for the Cy3 dye.

Special Notes and Considerations: This is a very stable and bright dye that is ideal for probe sets with smaller numbers of probes (less than 30) or for use in tissue as auto-fluorescence tend to decrease with longer wavelengths.

Anti-fade Recommendations: The Quasar 570 dye performs best with an anti-fade mounting media and is compatible with Vectashield by Vector Labs (H-1000), Prolong Diamond by ThermoFisher (P36961), Prolong Gold by ThermoFisher (P36930), and Slow Fade Diamond by ThermoFisher (S36963).

How long will my sample last after mounting?: When stored at +2 to +8 °C in the dark, the Quasar 570 signal remains stable for up to a week in the anti-fades listed above. Hardmount anti-fades such as the Prolong products and Vectashield soft set may extend the life of your sample for months past mounting.

Filter Recommendation: Our in-house team uses Chroma's SP102v1-Cy3™ v1 single band filter set. Chroma's 49304-ET-Gold FISH single band filter also fits Quasar 570 well.

Absorption and Emission Spectra for Quasar 570 Dye

Properties:

• Name: Quasar 570
• Formula: C₃₄H₄₅N₃O₃
• Absorption maximum (lambda max): 548 nm
• Extinction coefficient at lambda max: 115,000 M^-1cm^-1
• Extinction coefficient at 260 nm: 9,000 M^-1cm^-1
• Fluorescence maximum: 566 nm

Design considerations

Please read the tech article titled, “Designing Stellaris RNA FISH Probes” before designing your custom assay.

You can also use Chroma’s Spectra Viewer to check your microscope’s dye filter set compatibility. Use the Stellaris Dyes and Modifications page to select the right dye for your microscope and multiplexing recommendations.

Note I: Due to the variable autofluorescence in the green channel inherent to some cells and tissues, it is not recommended to use FAM for some Stellaris RNA FISH assays. We recommend testing your sample for green channel autofluorescence before ordering FAM labeled sets or ordering an alternative dye that is compatible with your experiment.

Note II: The success rate of Custom Probe Sets depend on many variables, such as imaging platform, operator use of optics, target sequence, transcript number, subcellular localization of transcript, number of probes in the set, space between probes, masking level, sample identity whether cultured cells or fresh or frozen tissue, preparation and handling of sample, and customer's design of probe sets with or without the Stellaris Probe Designer. The customer is responsible for designing a probe set that is compatible with all of the variables mentioned above unique to their platform technologies and model system, and therefore we cannot guarantee the success of Customer Probe Sets, or troubleshoot the designs.

Please contact us by email, phone or using this form for further advise and details.