ViewRNA™ Tissue Fluorescence 4 Plex Assay Kit (green, red, deep red, and near infared) - FAQs

View additional product information for ViewRNA Tissue Assay Fluorescence Kits - FAQs (QVT0646B, QVT4800, QVT4700, QVT0600C, QVT0688B, QVT4646B, QVT4640B, QVT4694B, QVT4688B, QVT4600C, QVT0800, QVT0700, QVT0694B, QVT0640B)

11 product FAQs found

How do ViewRNA assays compare to RNAScope assays?

ViewRNA and RNAScope technologies rely on the same signal amplification strategy - branched DNA amplification. Historically, both ViewRNA and RNAScope technologies originated from the same company, Panomics. The assays are expected to yield similar sensitivity and resolution, however each technology relies on its own set of proprietary reagents and probe set designs. Hence, the assays are not considered interchangeable or compatible. ViewRNA probe sets are not tested for RNAScope assaya and vice versa.

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How is the signal amplified in ViewRNA assays?

The ViewRNA technology relies on branched DNA signal amplification strategy. Target probes complementary to the target transcript sequence are further hybridized with pre-amplifier, amplifier and label probes that consist of branched DNA, and form 'tree branches' that allow numerous label probes to attach. This approach allows higher signal amplification compared to traditional ISH techniques.

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Where can I find general information about ViewRNA ISH Assays?

For general information about ViewRNA ISH Assays, please go to this page (https://www.thermofisher.com/us/en/home/life-science/cell-analysis/cellular-imaging/in-situ-hybridization-ish/rna-fish/viewrna-assays.html).

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For ViewRNA assays, can I use the same set of wash solutions for all samples, including the positive and negative controls?

We do not recommend doing this. The negative control should be processed and washed separately from the rest of the samples. This is because the negative control does not contain any target probe sets and only the amplification reagents are added to it. If experimental samples are washed in the same beaker of wash solutions as the negative control, any unbound target probes that wash away can carry over to the negative control sample and cause unexpected positive signal (that will also appear to be very specific).

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Is there a video on the ViewRNA tissue assay workflow that I can watch?

Yes, we do have a video that will help provide hands-on guidance on how to proceed with the protocol. Please find the link to the online protocol and video (https://www.thermofisher.com/us/en/home/references/protocols/cell-and-tissue-analysis/protocols/viewrna-tissue-fluorescence-protocol.html).

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Do you have a list of antibodies that are compatible with ViewRNA assays?

Unfortunately, we do not have such a list as each antibody may perform differently on the varied sample types. We recommend testing each antibody in the context of the ViewRNA assay workflow to ascertain quality results. Assay conditions can be optimized by modifying the protease treatment time as well as optimizing heat pretreatment for antigen recovery.

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Do you have a list of tissue types tested to work with ViewRNA Tissue Assay kits?

Tissue types that have been tested to work with ViewRNA Tissue Assay kits can be found listed on page 30 of the ViewRNA Tissue Fluorescence Assay User Guide (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0026643_ViewRNA_TissueFluorAssay_UG.pdf).

Tissue types that have been tested to work with ViewRNA Tissue Assay kits can be found listed on pages 29-30 of the ViewRNA Tissue Fluorescence Assay User Guide (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0026643_ViewRNA_TissueFluorAssay_UG.pdf) and on pages 48-49 of the ViewRNA Tissue Colorimetric Assay User Guide (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0018633_viewRNA_ISH_UG.pdf).

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Can I multiplex ViewRNA ISH tissue fluorescence assays with IHC or IF (antibody immunostaining)?

It is possible to combine RNA ISH on tissues with IF or IHC using immunofluorescence. It is essential to test the antibodies on the desired tissue type and under the conditions of the ViewRNA tissue assay workflow. Due to variable tissue processing conditions, not all antigens may remain intact. Note that antibodies added to the ViewRNA workflow should be RNAse-free or RNAse inhibitor-treated. Guidelines for combining RNA ISH and IHC can be found on page 31 of the ViewRNA Tissue Fluorescence Assay User Guide (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0026643_ViewRNA_TissueFluorAssay_UG.pdf).

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Can I use the reagents from ViewRNA Tissue Assay Fluorescence kits and ViewRNA Colorimetric Tissue Assay kits interchangeably?

We do not recommend interchanging reagents between ViewRNA Tissue Assay Fluorescence kits and ViewRNA Colorimetric Tissue Assay kits. While some reagents, such as the Wash buffer components can be used interchangeably, other components such as the amplifier reagents are not formulated the same way.

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Are all ViewRNA Tissue Assay kits capable of performing up to a 4-plex assay?

No, the ViewRNA Colorimetric Tissue Assay kits only allow up to a 2-plex assay. To perform a 2-plex colorimetric tissue assay, the ViewRNA Tissue Core Kit (Cat. No. QVT0400) can be used in combination with ViewRNA Modules purchased separately (Cat. No. QVT0410, QVT0420, QVT0430) or one of the ViewRNA Colorimetric Tissue 2-Plex kits can be used (Cat. No. QVT4433C, QVT0433C, QVT0412C, QVT4412C, QVT0422C, QVT4422C).

ViewRNA Tissue Assay Fluorescence kits allow up to a 4-plex assay with purchase of kit Cat. No. QVT0700, QVT4700, QVT0800, or QVT4800 or by using the ViewRNA Tissue Fluorescence 1 Plex Assay Core Kit (Cat. No. QVT0600C or QVT4600C) in combination with the pertinent Type 1 (Cat. No. QVT0646B, QVT4646B, QVT0694B, QVT4694B), Type 4 (Cat. No. QVT0688B, QVT4688B), or Type 10 (Cat. No. QVT0640B, QVT4640B) ViewRNA Tissue modules purchased separately.

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What ViewRNA ISH assay kits do you offer for tissue samples?

ViewRNA ISH tissue assays are offered in two versions: colorimetric and fluorescence RNA ISH tissue assays.

- ViewRNA Tissue Assay Fluorescence Kits come with Alexa Fluor dye-conjugated label probes and enable the simultaneous detection of one to four RNA targets (mRNA, non-coding RNA, or microRNA) at single-copy sensitivity (Cat. No. QVT0700, QVT4700, QVT0800, QVT4800, QVT0600C, QVT4600C, QVT0688B, QVT4688B, QVT0646B, QVT4646B, QVT0694B, QVT4694B, QVT0640B, QVT4640B).

- ViewRNA Colorimetric Tissue Core Kit and Individual Modules rely on enzyme-mediated signal amplification, using AP- or HRP-conjugated label probes in conjunction with Fast Red, Fast Blue, or DAB substrates. These colorimetric assays enable detection of one or two mRNA targets within tissue sections at single-copy sensitivity (Cat. No. QVT0410, QVT0420, QVT0430, QVT0400, QVT0400A, QVT0400B).

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