Collaborative work between researchers at the IBPM platform and CrestOptics Company allowed the validation of a method for automated imaging and analysis of in vitro 3D breast epithelial acini. 

The derived Application Note (https://crestoptics.com/high-content-imaging-for-morphological-profiling-of-3d-breast-acini/) describes high-content platforms, based on the CrestOptics X-Light V3 spinning disk, applied to the morphological profiling of 3D breast spheroids derived from the MCF10A cell line.

The imaging system, coupled with Nis Elements Software capable of automating acquisitions and analyses, has made it possible to compare morphological characteristics between normal (MCF10A CTR) and transformed (MCF10A RAS) acini. High content analysis of morphological features such as volume, elongation, sphericity, diameter, and surface demonstrated that acini derived from transformed cells display an alteration of the acinar structure and architecture (see figure).

The combination of high content microscopy with tridimensional cellular models could thus contribute significantly to cancer research, especially in the validation of active molecules in drug screening.

Images obtained at the Imaging platform were presented at the Exhibition on Scientific Photorgraphy “Molecole”, organized by Associazione Giovanile Laboratorio11 in response to the call  “Vitamina G”, in collaboration with the  Dept of Biology and Biotechnology Charles Darwin (BBCD), the PhD course  in Genetics and Molecular  Biologiy of Sapienza University of Rome, the BBCD Department Library, and the  inter-departimental Centre  “Saperi&Co” of  Sapienza. 

13-14 October 2022, Container, via degli Etruschi, 15 Roma

https://bbcd.bio.uniroma1.it/bbcd/archivionotizie/molecole-13-e-14-ottobre-degli-etruschi-n-5-san-lorenzo-dalle-ore-1800-mostra

The project “Metodologie innovative di 3D high content imaging per la validazione di nuove molecole antitumorali” coordinated by Dr Francesca Degrassi has been funded by Regione Lazio within the Call “Progetti Gruppi di Ricerca 2020”.

Polverino F, Naso FD, Asteriti IA, Palmerini V, Singh D, Valente D, Bird AW, Rosa A, Mapelli M, Guarguaglini G. The Aurora-A/TPX2 Axis Directs Spindle Orientation in Adherent Human Cells by Regulating NuMA and Microtubule Stability.

Curr Biol. 2021. 31:658-667.e5.

doi: 10.1016/j.cub.2020.10.096. Epub 2020 Dec 3.

In this study we characterised the role of the Aurora-A kinase and its major regulator TPX2 in the control of mitotic spindle orientation in mammalian cells, a process which determines the position and fate of daughter cells.

Work was collaboratively carried out by groups at the Institute of Molecular Biology and Pathology (IBPM) and at the European Institute of Oncology (IEO). Research groups at Sapienza University and at the Max Planck Institute of Molecular Physiology (Dortmund, DE) participated to the project.

IBPM reference person: Giulia Guarguaglini

Artificial Intelligence-based analysis of commitment towards neuronal differentiation in living pluripotent cells over time

Neuronal differentiation is a highly complex, multistep process that regulates the determination and specification of neuronal fate, as well as major molecular and functional changes, such as the emergence of neurites and maturation of dendrites and axons. Acquiring a profound knowledge of factors regulating neuronal differentiation is crucial to identify particularly vulnerable windows in which defects or gene mutations can cause neurodevelopmental disorders. This requires capturing the dynamic dimension of the process.  

Here we aimed at imaging neuronal differentiation over time, both in fixed cell cultures and under in vivo conditions, to depict morphological cell changes and appearance of neuronal markers. We first characterized differentiating cell models exposed to classical inducers, i.e. Retinoic Acid (RA) and Neurobasal (NB) Medium. We next selected murine P19 pluripotent cells undergoing differentiation for time-lapse recording (up to 96 h) to follow up in real time their commitment to the neuronal fate. We developed two novel artificial intelligence (AI)-based algorithms to perform a high-throughput analysis of the phenotypes elicited during neuronal differentiation.

https://vizbi.org/2023/

A theoretical-practical course was offered at the Imaging platform for PhD students from 9 PhD courses associated to the PhD School in Molecular Biology and Medicine at Sapienza University of Rome.

An account of how coupling high-resolution imaging metods with mathematics and physical modelling provides insight into cell dynamics.

Guest speaker: Emanuele Roscioli, Fondazione Toscana Life Sciences, Siena.

Euro-BioImaging is the European landmark research infrastructure for biological and biomedical imaging. Through Euro-BioImaging, life scientists can access imaging instruments, expertise, training opportunities and data management services. The webinar presented the activity of the Italian nodes to CNR biomedical scientists.

Cellular imaging is undergoing an unprecedented development that has greatly amplified its informative power with the introduction of machine learning, artificial intelligence-based approaches and quantitative data analysis. The workshop has explored the application of these methods in as diverse cell biology fields as cell migration a, morphological reconstruction of subcellular structures, and use of model organism- and cell-based assays in genetic and drug screening.