Immunotherapy efficacy relies on the crosstalk within the tumor microenvironment between cancer and dendritic cells (DCs) resulting in the induction of a potent and effective antitumor response. DCs have the specific role of recognizing cancer cells, taking up tumor antigens (Ags) and then migrating to lymph nodes for Ag (cross)-presentation to naïve T cells. Interferon-α-conditioned DCs (IFN-DCs) exhibit marked phagocytic activity and the special ability of inducing Ag-specific T-cell response. Here, we have developed a novel microfluidic platform recreating tightly interconnected cancer and immune systems with specific 3D environmental properties, for tracking human DC behaviour toward tumor cells. By combining our microfluidic platform with advanced microscopy and a revised cell tracking analysis algorithm, it was possible to evaluate the guided efficient motion of IFN-DCs toward drug-treated cancer cells and the succeeding phagocytosis events. Overall, this platform allowed the dissection of IFN-DC-cancer cell interactions within 3D tumor spaces, with the discovery of major underlying factors such as CXCR4 involvement and underscored its potential as an innovative tool to assess the efficacy of immunotherapeutic approaches.

Mammography is the most effective method for the early detection of breast diseases. However, the typical diagnostic signs such as microcalcifications and masses are difficult to detect because mammograms are low-contrast and noisy images. In this paper, a novel algorithm for image denoising and enhancement based on dyadic wavelet processing is proposed. The denoising phase is based on a local iterative noise variance estimation. Moreover, in the case of microcalcifications, we propose an adaptive tuning of enhancement degree at different wavelet scales, whereas in the case of mass detection, we developed a new segmentation method combining dyadic wavelet information with mathematical morphology. The innovative approach consists of using the same algorithmic core for processing images to detect both microcalcifications and masses. The proposed algorithm has been tested on a large number of clinical images, comparing the results with those obtained by several other algorithms proposed in the literature through both analytical indexes and the opinions of radiologists. Through preliminary tests, the method seems to meaningfully improve the diagnosis in the early breast cancer detection with respect to other approaches.

In this paper we discuss the applicability of numerical descriptors and statistical physics concepts to characterize complex biological systems observed at microscopic level through organ on chip approach. To this end, we employ data collected on a microfluidic platform in which leukocytes can move through suitably built channels toward their target. Leukocyte behavior is recorded by standard time lapse imaging. In particular, we analyze three groups of human peripheral blood mononuclear cells (PBMC): heterozygous mutants (in which only one copy of the FPR1 gene is normal), homozygous mutants (in which both alleles encoding FPR1 are loss-of-function variants) and cells from ‘wild type’ donors (with normal expression of FPR1). We characterize the migration of these cells providing a quantitative confirmation of the essential role of FPR1 in cancer chemotherapy response. Indeed wild type PBMC perform biased random walks toward chemotherapy-treated cancer cells establishing persistent interactions with them. Conversely, heterozygous mutants present a weaker bias in their motion and homozygous mutants perform rather uncorrelated random walks, both failing to engage with their targets. We next focus on wild type cells and study the interactions of leukocytes with cancerous cells developing a novel heuristic procedure, inspired by Lyapunov stability in dynamical systems.

Microfluidic devices are increasingly widespread in the literature, being applied to numerous exciting applications, from chemical research to Point-of-Care devices, passing through drug development and clinical scenarios. Setting up these microenvironments, however, introduces the necessity of locally controlling the variables involved in the phenomena under investigation. For this reason, the literature has deeply explored the possibility of introducing sensing elements to investigate the physical quantities and the biochemical concentration inside microfluidic devices. Biosensors, particularly, are well known for their high accuracy, selectivity, and responsiveness. However, their signals could be challenging to interpret and must be carefully analysed to carry out the correct information. In addition, proper data analysis has been demonstrated even to increase biosensors' mentioned qualities. To this regard, machine learning algorithms are undoubtedly among the most suitable approaches to undertake this job, automatically learning from data and highlighting biosensor signals' characteristics at best. Interestingly, it was also demonstrated to benefit microfluidic devices themselves, in a new paradigm that the literature is starting to name "intelligent microfluidics", ideally closing this benefic interaction among these disciplines. This review aims to demonstrate the advantages of the triad paradigm microfluidics-biosensors-machine learning, which is still little used but has a great perspective. After briefly describing the single entities, the different sections will demonstrate the benefits of the dual interactions, highlighting the applications where the reviewed triad paradigm was employed.

We describe a novel method to achieve a universal, massive, and fully automated analysis of cell motility behaviours, starting from time-lapse microscopy images. The approach was inspired by the recent successes in application of machine learning for style recognition in paintings and artistic style transfer. The originality of the method relies i) on the generation of atlas from the collection of single-cell trajectories in order to visually encode the multiple descriptors of cell motility, and ii) on the application of pre-trained Deep Learning Convolutional Neural Network architecture in order to extract relevant features to be used for classification tasks from this visual atlas. Validation tests were conducted on two different cell motility scenarios: 1) a 3D biomimetic gels of immune cells, co-cultured with breast cancer cells in organ-on-chip devices, upon treatment with an immunotherapy drug; 2) Petri dishes of clustered prostate cancer cells, upon treatment with a chemotherapy drug. For each scenario, single-cell trajectories are very accurately classified according to the presence or not of the drugs. This original approach demonstrates the existence of universal features in cell motility (a so called “motility style”) which are identified by the DL approach in the rationale of discovering the unknown message in cell trajectories.