Periodic Reporting for period 1 - SameMultiPhys (Novel Biophysical Tools to Measure Multiple Parameters In The Same Cell)
Periodo di rendicontazione: 2023-11-01 al 2025-10-31
Mechanobiology is a dynamic and rapidly evolving interdisciplinary field focused on understanding how physical forces influence cells and tissues at multiple scales. This field holds immense potential for advancing healthcare and diagnostics, but its progress relies on the development of precise tools and methodologies capable of applying and measuring mechanical forces across biological systems. Despite significant advancements, there remains a critical gap in technology: the ability to probe multiple biophysical parameters—such as internal ordering, mechanical properties, electrical characteristics, and structural features—within the same cells. Addressing this challenge is essential for gaining deeper insights into cellular behavior variability, potentially unlocking new diagnostic and therapeutic possibilities.
The SameMultiPhys project proposes to develop innovative microfluidic technologies for the field of cell mechanobiology. These technologies will enable the measurement of multiple biophysical parameters on the same cells, offering a powerful tool for identifying biomarkers that reflect cellular states.Funded for four years under the Marie Skłodowska-Curie Actions Staff Exchange program, the project fosters international collaboration by connecting researchers from eight institutions through a series of research exchanges. Over its duration, the project will facilitate 39 one-month research exchanges across prestigious institutions, including Tel Aviv University, the University of Florence, CNRS, New York University, the University of Washington, the University of Santiago de Chile, and the University of Toronto.
The goals of the project are: (i) To design and develop new microfluidics-based technologies to perform biophysical assays capable of measuring multiple parameters at the single-cell level. This advancement aims to improve the identification of robust biomarkers for clinical applications and (ii) to validate and apply the developed tools in studies of T lymphocytes in various states of activation and senescence. These studies aim to unravel the complex biological processes underlying the immune response during aging. The specific Work Packages (WP) of the project are: WP1. Management; WP2. Designing and fabricating prototypes; WP3. Stardardizing measuremens with suitable calibration samples; WP4. Testing and validating prototypes; WP5. Quantifying acquired images and performing statistical analyses; WP6. Refine: Evaluating the prototypes for potential improvements; WP7. Development of biophysical models of cell response; WP8. Dissemination and communication.
The SameMultiPhys project proposes to develop innovative microfluidic technologies for the field of cell mechanobiology. These technologies will enable the measurement of multiple biophysical parameters on the same cells, offering a powerful tool for identifying biomarkers that reflect cellular states.Funded for four years under the Marie Skłodowska-Curie Actions Staff Exchange program, the project fosters international collaboration by connecting researchers from eight institutions through a series of research exchanges. Over its duration, the project will facilitate 39 one-month research exchanges across prestigious institutions, including Tel Aviv University, the University of Florence, CNRS, New York University, the University of Washington, the University of Santiago de Chile, and the University of Toronto.
The goals of the project are: (i) To design and develop new microfluidics-based technologies to perform biophysical assays capable of measuring multiple parameters at the single-cell level. This advancement aims to improve the identification of robust biomarkers for clinical applications and (ii) to validate and apply the developed tools in studies of T lymphocytes in various states of activation and senescence. These studies aim to unravel the complex biological processes underlying the immune response during aging. The specific Work Packages (WP) of the project are: WP1. Management; WP2. Designing and fabricating prototypes; WP3. Stardardizing measuremens with suitable calibration samples; WP4. Testing and validating prototypes; WP5. Quantifying acquired images and performing statistical analyses; WP6. Refine: Evaluating the prototypes for potential improvements; WP7. Development of biophysical models of cell response; WP8. Dissemination and communication.
The project's technical and scientific activities during the first reporting period (Months 1-24) align with the objectives outlined in Work Packages 2, 3, 4 and 5:
Work Package 2 (WP2, Months 2–34): We conducted an analysis of existing technologies for measuring multiple parameters in the same cell and developed several types of microfluidic devices capable of assessing both deformability and electrical properties. These devices are designed to accommodate a wide range of cellular sizes. Additionally, we implemented an experimental protocol and a custom-built analysis pipeline for the automated analysis of electrical and deformability measurements. The experimental assay simultaneously tracks: (a) the cell's aspirated length and area as a function of applied differential pressure, and (b) the cell's electrical impedance as a function of applied electric potential differences.
Work Package 3 (WP3, Months 13–24): We have begun defining the composition of calibration samples, ensuring they will be compatible with the microfluidic devices for use during upcoming secondments. The required materials are readily available, enabling preparation for the next stages.
Work Package 4 (WP4, Months 13–47): We developed novel assays for using microfluidic devices in biological contexts and initiated discussions with our partners to design experiments. These experiments aim to demonstrate the versatility of the devices across various cell lines.
Work Package 5 (WP5, Months 3–48): We have focused on automating time-lapse image analysis in microfluidic devices with constrictions. This includes estimating the number of blocked channels and tracking the aspirated length and area of cells in response to applied differential pressure.
Work Package 2 (WP2, Months 2–34): We conducted an analysis of existing technologies for measuring multiple parameters in the same cell and developed several types of microfluidic devices capable of assessing both deformability and electrical properties. These devices are designed to accommodate a wide range of cellular sizes. Additionally, we implemented an experimental protocol and a custom-built analysis pipeline for the automated analysis of electrical and deformability measurements. The experimental assay simultaneously tracks: (a) the cell's aspirated length and area as a function of applied differential pressure, and (b) the cell's electrical impedance as a function of applied electric potential differences.
Work Package 3 (WP3, Months 13–24): We have begun defining the composition of calibration samples, ensuring they will be compatible with the microfluidic devices for use during upcoming secondments. The required materials are readily available, enabling preparation for the next stages.
Work Package 4 (WP4, Months 13–47): We developed novel assays for using microfluidic devices in biological contexts and initiated discussions with our partners to design experiments. These experiments aim to demonstrate the versatility of the devices across various cell lines.
Work Package 5 (WP5, Months 3–48): We have focused on automating time-lapse image analysis in microfluidic devices with constrictions. This includes estimating the number of blocked channels and tracking the aspirated length and area of cells in response to applied differential pressure.
The SameMultiPhys project has demonstrated significant academic and research impacts, during the first period of the project, through the development of advanced microfludic devices capable of measuring both (i) the mechanical properties of individual cells, taking into account relative-size effects, based on the quasi-linear viscoelasticity theory, and (ii) the electrical properties of individual cells, based on impedance measurements.The methodology developed in this work opens the possibility for reliable individual-cell mechanical and electrical measurements measurements, providing a very useful tool to leverage the potential of these devices. Future works will apply the methodology to answer a variety of biological questions.