Optofluidic single-cell analysis platform

• Subject Area: Basic Research in Biology and Medicine ; Medicine
• Term: Funded in 2024
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 548189185

The device applied for is an analysis platform that enables high-dimensional examinations of individual cells and cell populations on a nano-chip. The device is novel, innovative and is intended to be used in pre-clinical and translational research projects for cellular immunotherapy of malignant and non-malignant diseases. The focus will be on research projects on chimeric antigen receptor (CAR)-modified T cells. The device works automatically and gently imports the cells to be examined using light pulses into nano chambers (pins) on a nano chip (10,000-20,000 pins/chip). The cells can be cultured in the nanochip under sterile conditions for several days and analyzed continuously. During import, the cells to be examined can be phenotyped and sorted using fluorescence-labeled surface markers, and then analyzed for functional parameters (e.g. CAR-T cells: proliferation and differentiation, cytokine pattern and sequence, cytotoxicity kinetics and mechanism, avidity/affinity and Cell-cell interaction with other immune cells or tumor cells) can be dynamically examined, observed (imaging) and modulated. Both immune and non-immune cells can be loaded into the chambers as individual cells or cell populations in different ratios depending on the desired experimental setup and analyzed in short- or long-term experiments (24 hours/1 day - up to 168 hours/7 days). The same cells can then be sorted and exported for further downstream analyzes at single-cell resolution (e.g. in vivo function, super-resolution microscopy, genomics, proteomics) based on the previously collected functional parameters. This analysis process consisting of phenotyping - sorting - high-dimensional functional analysis / imaging sorting and export for further analyzes cannot yet be reproduced with conventional devices and methods. With the proposed device, longitudinal, high-dimensional data (>100,000 data points on >30 parameters/cell in 7 days) can be collected for the first time on one and the same cell. Thanks to the innovative light pulse/pin technology, depending on the question, very small cell populations can be sufficient for data collection (e.g. from patient samples), but very large cell populations can also be processed (e.g. in screening experiments). At the Chair of Cellular Immunotherapy, the proposed device will primarily be used to address questions in the field of CAR-T cell immunotherapy in order to open up new possible applications in the area of malignant and non-malignant diseases that involve cell products to analyze and continuously improve their composition of defined subpopulations, their effectiveness, safety and production.

• DFG Programme: Major Research Instrumentation
• Major Instrumentation: Optofluidische Einzel-Zell-Analyse-Plattform
• Instrumentation Group: 3500 Zellzähl- und Klassiergeräte (außer Blutanalyse), Koloniezähler
• Applicant Institution: Julius-Maximilians-Universität Würzburg
• Leader: Professor Dr. Michael Hudecek