Introduction

The invention proposes a new computational method and the corresponding imaging system for the identification of subcellular structures, without the use of chemical stains, through an advanced analysis of 3D tomograms of cells acquired by means of a phase-contrast tomographic microscope in flow cytometry configuration. The proposed technology allows performance comparable to fluorescence-based imaging flow cytometry systems on the market, but with the great advantage of being completely green.

Technical features

The invention (TRL=4) focuses on a new computational method for the identification of sub-cellular structures without the use of chemical markers. The method is based on an advanced statistical analysis of tomographic images, obtained through the numerical processing of digital holograms of cells flowing and rotating in a microfluidic channel. Therefore, the acquisition and processing system is composed of three modules, i.e. (i) a microfluidic module, consisting of a microchannel in which a sample of cells (without chemical stains) is injected and pushed in continuous-flow through a microfluidic pump. Through the action of hydrodynamic forces, each cell is induced to rotate; (ii) a holographic microscope, suitably designed to ensure the highest possible resolution combined with a wide field of view, for the label-free imaging of cells; (iii) finally, the acquired holographic data are processed to reconstruct the volumetric distribution of the refractive index (phase-contrast tomogram) of each cell from which the analysis of intracellular specificity can be performed.

Possible Applications

• Statistical studies about cellular populations;
• Detecting circulating tumor cells in human body fluids for the early diagnosis of cancer (liquid biopsy);
• Rare diseases diagnosis;
• Testing the effectiveness of new therapies;
• Studying the cellular effects induced by external agents (e.g., nanoparticles, ionizing radiations).

Advantages

• Green technology;
• Non-invasive and potentially high-throughput imaging;
• 3D imaging (phase-contrast tomography);
• Measurements of morphological and biophysical features at single-cells level;
• Organelles characterization (intracellular specificity);
• Technology with lower provisional cost than competing systems on the market.