Innovative Uses of RNAscope

Read about reseachers who take innovative approaches with RNAscope® protocols to advance their research. If you would like to try one of these customer-demonstrated applications, please note that they are non-standard applications and therefore, a careful read through of the publications/materials is highly recommended. Our technical support team is here and will make best efforts to support you.

Customer demonstrated application:
Researchers Successfully Apply RNAscope ISH to 25-year-old FFPE Samples

Innovations in the Spotlight:

Erasmus MC Researchers Successfully Applied RNAscope® ISH to 25 year old FFPE Samples

 

You may be surprised to know that your archived FFPE samples may not be too old for RNAscope® in situ hybridization (ISH)! Our R&D team routinely detects RNA transcripts in three year old FFPE tissue specimens and some of our customers have applied RNAscope to much older tissues.  Read below, data from one of our customers where RNAscope ISH was successfully used in tissue specimens older than 25 years.

Success of RNAscope® with older FFPE samples depends on many factors including sample fixation, quality and storage. ACD cannot guarantee results for samples outside of our protocol recommendations. Please read FAQs for RNAscope® assay on archived FFPE samples section below for more information.

ACD Customers at Erasmus MC Push the Limits within RNAscope® FFFPE Protocol

Researchers Marije Hoogland and Esther Verhoef (Department of Pathology, Erasmus MC Rotterdam, the Netherlands) have conducted a retrospective study and successfully performed RNAscope® in situ hybridization on samples containing human metastases of prostate cancer in a lymph node. The samples were collected in 1987, 1988 and 1989 and analyzed in 2014 with RNAscope® probe targeting UBC gene. At 25 to 27 years old, these are the oldest samples we know of that have been used with RNAscope® in situ hybridization. The team followed ACD protocols and user manuals.

FAQs for RNAscope® Assay on archived FFPE samples

How can I ensure that my FFPE tissue specimens will work with RNAscope® FFPE Assay?

We have conducted extensive assay development to make sure that the RNAscope® FFPE assay will work with a wide range of tissue types.

  • If your tissue specimens are processed according to the Sample Preparation Guide in the User Manual and they are prepared within the last 3 years, you should expect successful staining for positive control probe using our assay procedure.
  • If your specimens are prepared differently from our recommended procedure, RNAscope® can still be used but may require assay optimization for some procedural steps. Please refer to our getting started page for protocol and sample guidelines and optimization.
  • Ensure that the assay is run according to RNAscope® protocol.

What are the recommended RNAscope® sample storage and pretreatment conditions for FFPE samples?

ACD sample storage requirements can be found in our RNAscope® Sample Preparation and Pretreatment Guide for FFPE Tissue, PART 1. The recommended pretreatment conditions for FFPE samples are based on tissue that has been fixed for 16-32 hrs at room temperature in 10% NBF.

What should I do if I don’t know whether the FFPE tissue specimen is appropriately fixed and processed?

Determine your samples' RNA quality following optimization recommendations on our getting started page (www.acdbio.com/go).

Figure 1: Punctate red dots show expression of UBC gene in Human prostate cancer samples from lymph node. RNAscope Chromogenic Red assay applied to FFPE  sample from 1989 , 400x magnification.

Figure 2: Expression of UBC gene (shown as punctate red dots) in human prostate cancer samples from lymph node. RNAscope Chromogenic Red Assay applied to FFPE samples from 1989, 400x magnification.

Figure 3: Expression of UBC gene (shown as punctate red dots) in human prostate cancer samples from lymph node. RNAscope Chromogenic Red Assay applied to FFPE samples from 1987, 400x magnification.

Figure 4: Expression of UBC gene (shown as punctate red dots) in human prostate cancer samples from lymph node. RNAscope Chromogenic Red assay applied to FFPE samples from 1989, 400x magnification.

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