RNAscope®/ BaseScope™/ miRNAscope™ Assays
RNA (예, probe)를 이용하여 조직 내 특정 RNA의 유전자 위치 및 변화 분석
- Any Gene, Any Species, Any Tissue
- 높은 민감도와 특이성: mRNA, lncRNA, Exon junction, point mutations 등 분석
- Multiplex: 한 시료에서 서로 다른 12개까지 유전자 발현 확인
- 분석 서비스 지원
Follow a step-by-step visual guide to assist with running the RNAscope Manual Assay.
How to use EZ-Batch Tray |
De-paraffinization |
Endogenous Peroxidase Blocking |
De-paraffinization is performed to ensure complete removal of the paraffin from FFPE samples to allow for the probes to penetrate the target RNA after adequate pretreatment. | RNAscope H2O2 step is performed during the RNAscope assay to block endogenous peroxidase enzyme activity to prevent hazy background after detection.Note: Pretreatment 1 refers to RNAscope H2O2 reagent and is the first pretreatment perfomed on your samples for Chromogenic assays. |
Target Retrieval (Boiling) |
Protease Plus (Protease Digestion) |
Target Probe Hybridization |
Target Retrieval step is a heat induced epitope retrieval method that is necessary to reverse the cross-linking caused by the formalin fixation step. Note: Pretreatment 2 refers to Target Retrieval reagent.For an alternative steamer protocol refer to the appendix of User Manual Part 2 Brown and Red. | Protease Plus is a broad spectrum protease that is intended to permeabilize the samples adequately to allow the probes to reach the target mRNA.Note : Pretreament 3 refers to Protease Plus reagent. | ACD provides properietary double “ZZ” oligo probes designed to hybridize to your specific RNA target |
Amplification (Amp1-Amp6) with Wash Step |
DAB Colormetric Reaction |
Gill’s Hematoxylin Counter Stain |
RNAscope detection reagents amplify hybridization signals via sequential hybridization of amplifiers. | Chromogenic detection is based on the enzyme substrate reaction which leads to the formation of an insoluble precipitate visualized in the form of punctate dots for the RNAscope assay. | Hematoxylin staining is a counterstain used to provide a contrast to better visualize the signal and to observe the morphology of the sample and identify the localization of the signal. |
Tissue Dehydration |
Mounting |
|
The tissue dehydration step after the counterstaining step results in removal of excess moisture which provides better tissue morphology and preservation of the signal. | The final step in RNAscope after staining requires the use of mounting media to adhere a coverslip to tissue section or cell smear. This helps protect the sample and the staining from physical damage and helps improve the clarity and contrast of an image during microscopy. |