RISH · Imaging of multiple mRNA targets using SERS nanoparticle labels and in situ hybridization in human cancer tissue sections
Horizon 2020 — Marie Skłodowska-Curie Actions
- Duration
- 2015-08-06 → 2017-08-05
- EU contribution
- €159,461
- Participants
- 1
- Scheme
- MSCA-IF
Lines connect the coordinator with its partners.
Results in brief
Imaging of multiple mRNA targets using SERS nanoparticle labels and in situ hybridization in human cancer tissue sections
Immunohistochemistry and in situ hybridization techniques that employ fluorescence are routinely used as diagnosis tools, for the detection of several antigens and DNA/RNA in tissue sections. Surface-enhanced Raman scattering (SERS), can also be employed in biomedical diagnostics. In immuno-SERS (iSERS) microscopy, the corresponding antibodies are labelled by molecularly functionalized noble metal colloids (SERS labels or nanotags). i.e. metal nanoparticles (NPs) with organic Raman reporter molecules chemisorbed on their surface. The reporter molecules provide the characteristic “molecular fingerprint” for the identification of the label, while the metal NP provides the necessary signal enhancement upon. During this project, SERS was employed for the detection of DNA biomarkers on breast cancer tissue sections. SERS applied for multiplex DNA and RNA detection for tissue diagnostics can have an impact on personalizing health and care. In situ hybridization utilizing Raman spectroscopy as an optical sensor has the potential of a quantitative multiplex in situ cancer diagnostic technique that can also provide spatial and cellular information. This level of insight can then lead to advances in early disease diagnosis, disease progression and response to therapy. Objectives: - Preparation and characterization of DNA targeting SERS probes. - Multiplex detection of DNA in FFPE breast tumour tissue sections. Optimization of tissue pre-treatment. - Detection of mRNAs in FFPE breast tumour tissue sections using Fluorescence and SERS microscopy
Data: CORDIS, © European Union
Project objective
Early cancer diagnosis greatly increases the chances for successful treatment and personalised therapy. RNA molecules have emerged as new candidate biomarkers for detecting a wide variety of cancers, due to their critical role in cell growth and development. In situ hybridization techniques for mRNA detection provide detailed spatial and temporal information on RNA expression with single-molecule sensitivity, however the traditional techniques that utilise fluorescence probes and quantum dots lack of high multiplexing capabilities. RISH seeks to develop a methodology that allows sensitive multiplex detection, quantification and distribution of cellular mRNA biomarkers utilising surface-enhanced Raman spectroscopy (SERS) nanoparticle (NP) labels. SERS NP-labels consist of metal nanoparticles modified with a self-assembled monolayer of Raman reporters and are sensitive and robust labels that offer great multiplexing capacity. Molecular beacon hybridization probes specific to mRNA targets will be conjugated to SERS NP-labels to allow optical detection utilising Raman microscopy in human cancer tissue sections. The mRNA expression levels will be correlated with those from fluorescence in situ hybridization and the respective protein expression levels after immunohistochemistry. The research proposal is highly multidisciplinary and includes aspects of spectroscopy, optics, biochemistry, nanotechnology and pathology. The research fellow plans to work closely with experts from both the hospital and the industry to apply this methodology in the diagnosis of initially breast cancer and of different types of cancer at a later stage.
Original text from CORDIS.
Participants
- UNIVERSITAET DUISBURG-ESSEN · EssenCoordinatorGermany
Links
Data: CORDIS, © European Union