Institute of Molecular Biology and Pathology
Visualizing cell dynamics
The IBPM microscopy platform hosts last-generation widefield light microscopy equipment and advanced imaging software for automated image analyses.
It supports versatile applications for both fixed and live cell samples and provides opportunities for coupling dynamic studies (time-lapse recording) with high resolution qualitative and quantitative image analysis.
The Center is open to external users, click here for access rules.
- Time-lapse recording of cellular processes in live cells over days
- Detecting intracellular molecular interactions
- Reconstructing cellular and subcellular structures at 3D level
- Quantitative analysis of imaging data
The platform was first established in 2009 with support from Fondazione Monte Paschi, Fondazione Roma and Assicurazioni Generali S.p.A. It has since been supported by two dedicated grants from Regione Lazio and by the CNR Project “Impara - Imaging from Molecules to Preclinical Studies” funded by the Ministry of Education and Research for the development of National research infrastructures. The platform is hosted at the Department of Biology and Biotechnology “Charles Darwin” of Sapienza University.
EMBO Workshop-VISUALIZING BIOLOGICAL DATA (VIZBI)
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.
High-content imaging for morphological profiling of 3D breast acini
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.
- Lia Asteriti
- Vincenzo Costanzo
- Francesca Degrassi
- Giulia Fianco
- Giulia Guarguaglini
- Patrizia Lavia
- Valerio Licursi
- Federica Polverino
- Davide Valente