Home › Microbiology › RNAscope for In situ Detection of Transcriptionally Active Human Papillomavirus in Head and Neck Squamous Cell Carcinoma
Microbiology JoVE (Open Access) Citable · DOI

RNAscope for In situ Detection of Transcriptionally Active Human Papillomavirus in Head and Neck Squamous Cell Carcinoma

DOI: 10.3791/51426-v
What you'll learn
  • ✓ Perform RNAscope in situ hybridization on FFPE tissue sections
  • ✓ Detect and visualize HPV E6/E7 mRNA in head and neck tumor samples
  • ✓ Interpret RNAscope assay results for HR-HPV transcriptional activity
Protocol

Biopharma Insights The presence of high-risk HPV in head and neck tumor tissue is associated with favorable outcomes. The recently developed RNA in situ hybridization technique called RNAscope allows direct visualization of HPV E6/E7 mRNA in FFPE tissue sections.

Difficulty
advanced
Total time
~4-6 hours per sample (assay protocol; tissue preparation time not included)
Biosafety
BSL-2

Steps

1
Prepare samples, equipment, and reagents

Gather FFPE tissue sections, RNAscope probe sets, detection reagents, and hybridization equipment. Prepare slides and work surfaces according to protocol specifications.

▶ 02:14
2
Execute RNAscope in situ hybridization assay

Perform stepwise RNAscope protocol on FFPE tissue sections, including probe hybridization, wash steps, and signal detection to visualize HPV E6/E7 mRNA.

▶ 04:07
3
Analyze RNAscope detection of HR-HPV

Examine stained tissue sections microscopically to identify and characterize HPV E6/E7 mRNA signal patterns in tumor tissue, confirming transcriptional activity.

▶ 08:15
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