Research Laboratories

The research laboratories of Bambino Gesù Children’s Hospital are dedicated to translational research; they are equipped with advanced technologies that support the scientific activities and the study of the biological basis of diseases, as well as their treatment.

GOALS

The main objectives of the research laboratories include:

• improve the knowledge of the etiology of pediatric diseases, to contribute to the identification of preventive, diagnostic, therapeutic and patient management strategies 
• encourage continuous exchange between the clinical and the scientific units the Hospital, to support the production of scientific results that can be rapidly transferred to the patient’s bed (translational research)
• contribute to the process of transformation, modernization and internationalization that are currently underway in the field of translational research in Lazio Region 
• act as a reference point for translational research in the Mediterranean Basin and in Europe 

 

THE ORGANIZATIONAL MODEL

The division of the spaces dedicated to the laboratories reflects the organizational model of the scientific activity, with is composed of 9 Research Areas, each one made of different Units, with their specific objectives and goals. With a surface of over 5,000 square meters, the laboratories are located in the 5 floors of the building of San Paolo Fuori le Mura, and in the dedicated areas of the other facilities of the Hospital. Furthermore, other platforms ensure cross-cutting services supporting the Hospital’s activities.

 

THE MAP

THIRD FLOOR: TRANSLATIONAL IMMUNOLOGY – CANCER IMMUNOLOGY – MOLECULAR GENETICS AND FUNCTIONAL GENOMICS – ZEBRAFISH PLATFORM

SECOND FLOOR: TRANSLATIONAL CYTOGENOMICS – METABOLIC DISEASES – CLINICAL GENETICS AND DYSMORPHOLOGY – SHARED PLATFORMS 

FIRST FLOOR: NEUROMUSCULAR DISEASES – GENODERMATOSIS – NEUROLOGICAL AND NEUROSURGICAL CONDITIONS – KIDNEY DISEASES – PREDICTIVE AND PREVENTIVE MEDICINE – IMMUNE-RHEUMATOLOGY – MICROBIOME – SHARED PLATFORMS

GROUND FLOOR: GENETICS AND EPIGENETICS OF PEDIATRIC CANCER – CELL AND GENE THERAPY IN HEMATOLOGICAL DISEASES – SHARED PLATFORMS – BIOBANK – PHARMACEUTICAL WORKSHOP – PATHOPHYSIOLOGY OF GROWTH PROCESS – CANCER GENE THERAPY – MOLECULAR GENETICS OF COMPLEX CHARACTERS – PREDICTIVE AND PREVENTIVE MEDICINE – IMMUNOLOGY DIAGNOSTICS – PATHOPHYSIOLOGY OF THE BONE TISSUE

BASEMENT: COMPLEX CELL THERAPIES – CANCER GEN THERAPY – CELL AND GENE THERAPY IN HEMATOLOGICAL DISEASES – GENETICS AND EPIGENETICS OF PEDIATRIC CANCER – BIOBANK – SHARED PLATFORMS

 

TECHNOLOGICAL PLATFORMS

The laboratories are equipped with state-of-the-art devices, that are used by researchers for their scientific activities. A substantial part of the equipment is made of next-generation devices, primarily dedicated to research in the field of molecular and cellular biology. The labs also include IT platforms for data processing, that facilitate the communication and data sharing between the different scientific networks.

Cytofluorimetry

• 4 cytofluorimeters for the simultaneous analysis of up to 20 parameters with high sensitivity and resolution, for the analysis of extremely small cell populations (microvesicles and exosomes).
• 2 cytofluorimeters/sorters for the simultaneous analysis of up to 20 parameters, capable of discriminating and separating particles of sub-micron size 
• Mass spectrometry and chromatography 
• Single quadrupole UHPLC/MS system for qualitative and quantitative high-throughput analysis of target molecules in complex mixtures, for the screening of specific compounds and for the confirmation of the molecular weights of the analytes
• GC/MS-SPME system, for the metabolomic analysis of complex matrices
• Two UHPLC/MS/MS QTRAP/LIT platforms, for quantitative and qualitative analysis with high linear sensitivity, dedicated to metabolomic and lipidomic investigations 
• A nano-LC/MS/MS TTOF platform and a micro-LC/MS/MS QTRAP/LIT platform, for qualitative investigations, fast profiling of specific analytes, and high-resolution quantitative workflows for metabolomics and proteomics applications
• A mass spectrometry platform with tribid architecture (quadrupole, ion trap and OrbitrapTM) and equipped with nano-LC, with multiple fragmentation technologies (CID, HCD, ETD and EThCD), for in-depth discovery analysis, for the characterization of the post-translational protein biosynthesis phases, and for the qualitative and quantitative analysis of molecular flows and processes

Genomics and cytogenetics

• New systems for fast real time qPCR analysis, equipped with array technology for the analysis of high-throughput gene expression, miRNA, and genotyping
• New Sanger sequencing technologies (up to 24 capillaries), with standardization and in-lane normalization of the resultants, associated with systems for real-time data quality assessment 
• New technologies for the preparation and analysis of high-throughput single cell gene expression, for the detection of genomic heterogeneity and clonal evolution, for the profiling of lymphocyte transcripts, for regulatory mechanisms and epigenetic heterogeneity (Assay for Transposase-Accessible Chromatin), as well as for libraries for exome and genome-wide sequencing
• Next Generation Sequencing systems for the study and analysis of the exome, the transcriptome, and the whole human genome, as well as for metagenomics applications 
• Cutting-edge high-resolution microarray scanner for SNP genotyping, structural variant analysis, genome-wide association studies (GWAS), and quantitative analysis of methylation sites within the human genome
• Digital microscopy platform for karyotype analysis using consolidated techniques, such as FISH, CGH, M-FISH, SKY FISH, and automatic metaphase search
• Illumina NovaSeq 6000 Platform for genome-wide analysis: in the diagnostic field, in particular for the molecular diagnosis of undiagnosed patients, and for the improvement of patient management and the prediction of drug response; in the research field, in studies on the functional and clinical relevance of mutations affecting non-coding regions of the human genome, as well as for the identification of new disease genes and of new molecular mechanisms underlying genetic diseases

 

Traditional and confocal microscopy

• Automation of the activities of tissue processing, immunohistochemical preparation and tissue immunofluorescence
• Laser microdissection system to isolate areas of tissue, single cells, and subcellular structures (e.g., chromosomes), used for the selection of samples for genomics, transcriptomics, proteomics and metabolomics studies, as well as for the investigation of in vivo cell cultures 
• Microscopy platform for cellular investigations, with live-cell imaging technology and software for the qualitative and quantitative analysis of images
• Digital platform for high-capacity automatic scanning and archiving of histological slides, both in brightfield and fluorescence mode
• Integrated cytometry and live cell imaging system, both in brightfield and fluorescence mode, for the growth tracking of cell population in adherence; cell count in suspension; morphological analysis and monitoring of the growth in embryos, colonies, spheroids, wound healing, post transfection/transduction cell analysis; iPSC reprogramming monitoring (fibroblast doubling, colony count, immunostaining by differentiation); morphological and morphometric analysis of 3D models also for small animals; differentiation of cell populations and co-cultures; chemotaxis; 2D and 3D migration and invasion; cytotoxicity analysis without the use of radioactive agents; cell viability, internalization and phagocytosis, apoptosis, and cell secretion analysis
• Microscopy platform for high-content screening applications (confocal spinning-disk; transmission and fluorescence technology), for 2D and 3D acquisitions for assay of fixed and live cells, of spheroids and organoids, of co-cultures for the analysis of protein-protein interaction and conformational changes, phenotypic fingerprinting, and live-imaging even of small animals
• Two confocal microscopy stations (TCS SP8X, FV3000) operating in galvo and resonant mode FCS, FCCS, FLIM and FLCS), one of which is also equipped with a white laser with integrated hardware-software system for the analysis of the lifetime of the fluorescence (FLIM), of the energy transfer by resonance (FRET) between molecules, of the fluorescence recovery after photobleaching (FRAP); all these methods enable the qualitative and quantitative investigation of the dynamics and interactions of single molecules within complex cell systems. An incubation system that maintains the conditions of temperature, humidity and CO2 allows for live-cell imaging experiments on cultured cells.