Invitrogen™ ViewRNA™ Cell Plus Assay Kit

Catalog No.	Quantity
88-190-0099	1 kit

Catalog No. 88-190-0099 Supplier Invitrogen™ Supplier No. 881900099

Description

The ViewRNA Cell Plus Assay is a novel assay that combines immunocytochemistry with ViewRNA technology, a proprietary fluorescent in situ hybridization and sequential branched-DNA amplification technique, to visualize both RNA with single-molecule sensitivity and protein in individual cells.

This assay enables simultaneous detection of up to three RNA targets in combination with immunophenotyping for cell surface or intracellular proteins using both indirect and direct immunocytochemistry to allow for detailed characterization of specific cell subpopulations. For simultaneous detection of a fourth RNA target, the ViewRNA ISH Cell 740 Module (Cat. No. QVC0200) can be combined with this kit. It allows analysis of an additional target in the 740 channel (AlexaFluor 750).

This ViewRNA Cell Plus Assay kit contains all the reagents needed to conduct the assay. Target Probe sets for genes of interest and antibodies are sold separately.

Reactivity/species
Probes can be designed for any species. These are the species most frequently used/tested: Armenian hamster, baboon, bovine, canine, cat, chicken, chimpanzee, cow, cynomolgus monkey, donkey, goat, golden Syrian hamster, guinea pig, hamster, horse, human, macaque, monkey, mouse, non-human primate, olive baboon, pig, pigtailed macaque, rabbit, rat, rhesus monkey, sheep

Reported application
Microscopy, immunocytochemistry

Specifications

• Detection Method: Primer-probe Detection
• Format: Kit
• For Use With (Application): Microscopy, Immunocytochemistry
• Product Type: RNA In Situ Hybridization Assay Kit
• Quantity: 1 kit

Frequently Asked Questions (FAQs)

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.

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.

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).

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).

To perform ViewRNA assays, which incubator oven should I use, Cat. No. QS0704 or QS0712?

Cat. No. QS0704 is a 120 V unit, for use in US/Canada region (https://www.thermofisher.com/order/catalog/product/QS0704).

Cat. No. QS0712 is a 220 V unit, for use in European region (https://www.thermofisher.com/order/catalog/product/QS0712).

What equipment do I need to perform ViewRNA assays?

Here is a list of equipment that would be needed to perform ViewRNA assays:
- Hybridization incubator oven capable of maintaining temperature at 40 ± 1 degrees C (Cat. No. QS0704 or QS0712)
- Humidity chamber (StainTray device, Cat. No. 44-0404)
- Fluorescent microscope with 20X and 40X objectives, appropriate filters, and monochrome camera
- Aspiration system for washing
- ViewRNA Temperature Validation Kit (Cat. No. QV0523)
- Optional cytospin for attaching suspension cells to a slide

To perform ViewRNA assays, do I need anything else besides ViewRNA probe sets and a ViewRNA assay kit?

Besides common laboratory reagents that are listed in the user manual, we also recommend purchasing a positive control probe set for assay validation (optional).

Additionally, the following items are needed to perform ViewRNA assays:

- Hybridization incubator oven capable of maintaining temperature at 40 ± 1 degrees C (Cat. No. QS0704 or QS0712)
- Humidity chamber (StainTray device, Cat. No. 44-0404)
- Fluorescent microscope with 20X and 40X objectives, appropriate filters, and monochrome camera
- Aspiration system for washing
- ViewRNA Temperature Validation Kit (Cat. No. QV0523)
- Optional cytospin for attaching suspension cells to a slide

What is the protocol for performing ViewRNA ISH Cell and ViewRNA Cell Plus assays in 96-well plate format?

Guidelines for performing the ViewRNA cell assay protocol in a 96-well plate format can be found on page 12 of the ViewRNA Cell Plus Assay User Guide (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/88-19000.pdf). The provided general guidelines for how to scale the reactions down can be adapted for the ViewRNA ISH Cell Assay (Cat. No. QVC0001).

What is the optimal stopping point for the ViewRNA Cell Plus Assay?

The optimal stopping point is after the sample fixation/ permeabilization step, prior to addition of the target probe or antibody. For exact details, see page 6 of the user manual (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/88-19000.pdf).

Can I use the ViewRNA ISH Cell Assay Kit and the ViewRNA Cell Plus Assay Kit reagents interchangeably?

It is possible to use some reagents from the ViewRNA ISH Cell Assay Kit and the ViewRNA Cell Plus Assay Kit interchangeably, such as the wash buffer components, probe diluents, and amplifier and label probes. However, for optimal performance of the kits, we recommend adhering to the recommended reagents from each kit.

What is the difference between the ViewRNA ISH Cell Assay Kit and the ViewRNA Cell Plus Assay Kit?

The ViewRNA ISH Cell Assay Kit (Cat. No. QVC0001) enables the simultaneous detection of one to four RNA targets (either mRNA or non-coding RNA) at single copy sensitivity and single cell resolution using fluorescence microscopes or high-content imagers. Note that for simultaneous detection of the fourth RNA target, the ViewRNA ISH Cell Assay Kit has to be used in conjunction with the ViewRNA 740 Module (Cat. No. QVC0200) that allows analysis of an additional RNA target in the 740 channel (Alexa Fluor 750).

The ViewRNA Cell Plus Assay Kit (Cat. No. 88-19000-99) does not use protease digestion in its workflow and thus enables the simultaneous detection of one to four RNA targets in combination with immunophenotyping for cell surface or intracellular proteins using both indirect and direct immunocytochemistry to allow for detailed characterization of specific cell subpopulations. Note that for simultaneous detection of a fourth RNA target, the ViewRNA Cell Plus Assay Kit has to be used in conjunction with the ViewRNA ISH Cell 740 Module (Cat. No. QVC0200) that allows analysis of an additional RNA target in the 740 channel (Alexa Fluor 750).

For Research Use Only. Not for use in diagnostic procedures.