Funding: CNR-UVR AMICO2_PoC, through Next Generation EU PoC 2022 – PNRR del MIMIT-UIBM M1 C2 I6 https://www.cnr.it/it/bando-uvr-amico-poc-2022
Duration: February 2024 – March 2025
Description: CITOM is a Proof-of-Concept project, aiming at the development of a imaging flow cytometry system for analyzing cells in continuous flow using holotomographic microscopy in a completely stain-free mode. In fact, state-of-the-art imaging flow cytometers employ fluorescence microscopy to identify and study intracellular organelles. However, fluorescence microscopy is intrinsically an indirect imaging method mediated by chemical agents, thus resulting in well-known drawbacks, such as photobleaching and phototoxicity. Avoiding staining will permit to access non-destructive, rapid and chemistry-free analysis in biomedicine. The CITOM apparatus and related computational method for the stain-free intracellular specificity, named Computational Segmentation based on Statistical Inference (CSSI) ((Italian patent n. 102021000019490, dep. 22/07/2021, Intern. PCT/IB2022/056625, dep. 19.07.2022) aim to bridge the gap between fluorescence and label-free imaging modalities in terms of intracellular specificity thanks to the possibility to identify subcellular organelles from 3D tomographic images without using chemical stains.
See for details
https://www.knowledge-share.eu/it/brevetti/specificita-intracellulare-senza-marcatori
Pirone, D. et al. Stain-free identification of cell nuclei using tomographic phase microscopy in flow cytometry. Nat. Photon. 16, 851–859 (2022). https://doi.org/10.1038/s41566-022-01096-7
Objectives and Expected Results: The main objective of CITOM is to design and implement a holotomographic imaging flow cytometer by integrating:
- Holographic microscope equipped with a high frame rate camera
- Optimized microfluidics module to enhance the throughput of the system
- Generalized CSSI method for identifying multiple subcellular organelles
In particular, the optimized combination of the holographic microscope and the microfluidic module, with the aim to increase the number of inspected cells per minute along with the enhancement of the quality and precision of the tomographic data, will be the main challenge of the project.
It is expected to raise the level of maturity of the technology from TRL 4 up to TRL 5.
Project Team: The ISASI Team is made up of Pasquale Memmolo (Scientific Leader of the project, Senior Researcher), Pietro Ferraro and Lisa Miccio (Research Director and Senior Researcher, respectively, with expertise in digital holography and tomography imaging), and Vittorio Bianco (Senior Researcher with expertise in image processing and computer vision).
Total Funding: 100.000 €