Simultaneously nanomechanical and fluorescence characterization of fresh tissue biopsies (MechanoFluorescence)

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Cancer progression is closely related to changes in mechano-cellular phenotype and in the structure and mechanical properties of the tumor microenvironment-TME in a complex and not well-understood manner. Although novel therapeutic strategies that target cancer cells or the TME are emerging, their efficacy varies due to intra- and inter-patient variability as well as tumor heterogeneity. Given that only some patients respond to a particular treatment, a new era of personalized, patient-specific treatments has been initiated, the basis of which is the identification of biomarkers that characterize a particular tumor. Our work so far has demonstrated that Atomic Force Microscopy-AFM techniques and collagen-based optical characterization can lead to the development of novel MechnanoOptical signatures that can be used as treatment monitoring and prognostic predictive biomarkers. However, a number of drawbacks limit the possible clinical use of the MechanoOptical signatures in real clinical practice. AFM nanomechanical characterization of tissue requires the experiments to be conducted within the same day of tissue harvest, while the majority of the techniques for assessing the collagen-based signatures require quite complex tissue preparation. Also, so far for combining collagen-based optical characterization with AFM mechanical signatures, expensive bio-AFM systems were used. In order to overcome these bottlenecks in this project, we proposed the development of the appropriate protocols for the simultaneously nanomechanical and fluorescence characterization of fresh tissue biopsies. A combination of cutting-edge experimental approaches in desmoplastic breast tumors will be used. Successful completion of the project will enable assessing the MechanoOptical signatures of fresh tissue specimens within the same day of tissue harvest leading to the possible clinical use of these novel biomarkers.

Lumoscribe was mainly responsible for the second Research Objective “RO.2. Develop protocol for staining and measuring fluorescence intensity from fresh tissue sections mounted on AFM system” and the fluorescence measurements.

Also we worked in collaboration with the HO members for achieving the third Research Objective “RO.3 Optimize protocol for simultaneously measurements of nanomechanical and fluorescence properties of fresh tissue specimens”.

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