Fluorescence and nanoscopy for early cancer diagnosis Early stage detection of breast and prostate cancer is crucial in order to further increase the survival rate of these patients. The EU funded a research initiative aimed at realising early breast and prostate cancer diagnosis using novel imaging and sampling methodologies that are safe and minimally invasive. Health © Thinkstock Core needle biopsy used for sample collection carries the risk of cancer seeding due to trauma from needle penetration at the tumour site. Although, fine needle aspiration (FNA) cell sampling is minimally invasive, less risky, faster and cost efficient, sample quantity is often too sparse for conclusive results. The FLUODIAMON project aimed to develop early cancer detection protocols that are safe and effective. This involved optimising the FNA sampling technique and image acquisition, resolution and specificity, identifying cancer biomarkers and standardising bioinformatics processing after validation. Researchers developed FNA sampling needles with better ultrasound visibility for accurate positioning. Penetration and manoeuvrability were improved by adding computer-controlled oscillations and bevelling the needle tip. The results were validated based on data from more than 200 breast cancer patients. A fully functional anti-seeding instrument with a specialised anti-seeding needle was also developed and validated on patients. Biomarkers found useful for the developed protocols and showing specific imaging features for breast and prostate cancer include vimentin and tubulin (cytoskeletal proteins), IGF1R, HER1, HER2 (membrane proteins), and Cyclin A and Cyclin E (cell cycle regulating proteins). Their spatial distribution patterns were studied in individual FNA sampled cells for the detection of malignant tumours. Stimulated emission depletion (STED) microscopy and multi-parameter fluorescence detection imaging (MFDi) methods were developed for the study of sub-cellular FNA samples. For this purpose, highly photostable and bright fluorophores were used as labels and coupled to relevant protein biomarkers in imaging procedures. Through bioinformatic quantitative analysis and verification, six independent diagnostic classifiers of malignant cancers were identified for their early detection. Knowledge gained was disseminated through 24 peer-reviewed articles, workshops and seminars, as well as on the website. Project members also applied for 10 specific patents. The results from this project have opened the doors for safer, patient-friendly, minimally invasive early stage diagnostic and therapeutic treatment of most cancers. These methodologies could also be adapted for follow up of pre-malignant lesions or evaluating disease progress and response to therapy.