The mechanical interplay between cell and the surrounding microenvironment is largely recognized to critically regulate cell function in tumor progression, malignancy transformation and metastasis but it remains poorly understood due to the lack of suitable measurement tools. While much progress has been achieved in the context of single-cell measurements of forces and deformation, measuring intracellular and extracellular moduli remains challenging, especially in 3D microenvironments. Yet, cell/ECM mechanical properties are known to be important because they link the presentation of an environmental mechanical stimulus to the activation of a mechano-related signaling pathway. In the past few years, we have been developing an all-optical approach to this challenge, named Brillouin microscopy. Brillouin cellular microscopy promises to map the intracellular and extracellular elastic modulus at high resolution, non-perturbatively, without contact in 3D microenvironments. As shown in strong preliminary data, we have achieved several key milestones that demonstrate our ability to measure relevant mechanical properties at a cellular and sub-cellular level. In this research we will develop and validate our microscopy platform for cancer-related studies.
Aim 1 will focus on the advanced development of the instrument to reach rapid mechanical imaging at sensitivities comparable to contact-based mechanical tests.
Aim 2 will focus on a direct validation of our technology against gold-standard techniques using properties and settings known to be relevant for metastatic progression. Finally, Aim 3 will test our technology within the context of tumor cell extravasation, a challenging experimental setting where no other technology can perform mechanical characterizations. Achieving sufficient speed and sensitivity for extravasation will guarantee the broad applicability of our technology for tumor mechanobiology and vividly demonstrate the type of novel information that our technology can provide. Importantly, Brillouin technology will be integrated into confocal microscopes to add a mechanical modality to widely-used instruments. The rigorous process of technology development, validation, benchmarking and field-testing will yield an instrumental platform with unprecedented capabilities to study cell-matrix biomechanics and ready to be widely adopted by the cancer biology research community.
This project will develop and validate Brillouin microscopy for high-resolution mapping of intracellular and extracellular matrix elastic modulus, properties known to be highly relevant in tumor biology. Brillouin microscopy is expected to provide new capabilities to characterize tumor progression and malignancy transformation as well as to identify which cell/ECM properties are crucial during the metastatic cascade. Integrated within widely-used confocal microscopes currently capable of only structural/functional imaging, our technology platform will enable widespread development of biomechanical studies in 3D microenvironments.
| Edrei, Eitan; Scarcelli, Giuliano (2018) Brillouin micro-spectroscopy through aberrations via sensorless adaptive optics. Appl Phys Lett 112:163701 |
| Zhang, Jitao; Nou, Xuefei A; Kim, Hanyoup et al. (2017) Brillouin flow cytometry for label-free mechanical phenotyping of the nucleus. Lab Chip 17:663-670 |
| Weber, Isabell P; Yun, Seok Hyun; Scarcelli, Giuliano et al. (2017) The role of cell body density in ruminant retina mechanics assessed by atomic force and Brillouin microscopy. Phys Biol 14:065006 |
| Edrei, Eitan; Gather, Malte C; Scarcelli, Giuliano (2017) Integration of spectral coronagraphy within VIPA-based spectrometers for high extinction Brillouin imaging. Opt Express 25:6895-6903 |
| Shao, Peng; Besner, Sebastien; Zhang, Jitao et al. (2016) Etalon filters for Brillouin microscopy of highly scattering tissues. Opt Express 24:22232-8 |
