In-flow tomographic imaging for single cells analysis

Abstract

Gold standard imaging modalities in biological field are based on fluorescence signals providing high specificity and high resolution. Recently, Fluorescence Microscopy has been combined with microfluidics to develop instrumentations called Imaging Flow Cytometers, high-throughput tools that supply bright-field, darkfield and multiple-channels fluorescence images of each single cell passing in the Field Of View (FOV). Nevertheless, Fluorescence Microscopy has some drawbacks as phototoxicity, photobleaching, expensive costs for sample preparations and also the a-priori knowledge of the tags to be used. For these reasons label-free imaging methods greatly increase in the recent years as the Quantitative Phase Imaging (QPI) technologies for microscopy. One of the optical techniques to achieve QPI is Digital Holography. DH in microscopy has several advantages such as the possibility to numerically scan the focal distance, a properties that open to the integration of DH in microfluidics. Indeed DH combined with microfluidic circuits allows to image particles or cells flowing into the FOV at different depths. Here the capabilities of label-free single-cell imaging by DH are presented and their implications on next future biomedical applications discussed. Static or in-flow configurations will be showed describing recent results and perspectives also in combination with Artificial Intelligence architectures for future applications in biomedical and clinical fields.