Gianluca Pegoraro, Ph.D.
- Center for Cancer Research
- National Cancer Institute
- Building 41, Room B909
- Bethesda, MD 20892-5055
Dr. Pegoraro heads the High-Throughout Imaging Facility (HiTIF), which provides CCR investigators with the expertise and the technology needed to set-up, optimize and implement high-content imaging (HCI) assays. HCI assays are used to analyze large numbers of experimental conditions, such as in functional genomics or chemical compounds screens. In addition, the HiTIF applies HCI in combination with single-cell image and data analysis to quantify exceedingly rare but biologically important events in heterogeneous populations of cells.
Areas of Expertise
1) high-content imaging 2) functional genomics 3) epigenetics
The High-Throughput Imaging Facility (HiTIF) is equipped with instrumentation and software for the automation of sample preparation, image acquisition and image analysis. Assays are generally run in either a 96- or 384-well imaging plates, and samples can either be fixed prior to image acquisition for end-point assays, or imaged live for time-lapse experiments. High-throughput image acquisition is performed on two Yokogawa spinning disk imaging platforms. Depending on the image acquisition parameters, the HiTIF output can reach up to 100,000 images per day. Custom image analysis routines are built and run using Columbus software on dedicated HiTIF workstations and/or on the Columbus image analysis server. In addition, we also take advantage of the scientific Python stack for custom-designed image analysis pipelines. The HiTIF staff is available to advise users on the experimental set-up and to assist and train them in every phase of the High-Content Imaging workflow.
More details regarding HiTIF instrumentation, publications, imaging fees, and the assay development process can be found at the HiTIF wiki site.
To ask for more information and to initiate project collaborations, please contact Dr. Pegoraro at the email address listed above.
Dynamic imaging of nascent RNA reveals general principles of transcription dynamics and stochastic splice site selection
Transcriptional Bursting and Co-bursting Regulation by Steroid Hormone Release Pattern and Transcription Factor Mobility
Gianluca Pegoraro, Ph.D.
Dr. Pegoraro received his M.Sc. degree in Molecular Biology from the University of Trieste, Italy in 2000. In 2004 he then obtained a Ph.D. degree in Molecular Genetics from the International School of Advanced Studies (ISAS), Trieste, Italy. Dr. Pegoraro joined the Cell Biology of Genomes Group (CBGE)/LRBGE/CCR as a Post-Doctoral Fellow in 2006. Here, under the supervision of Dr. Tom Misteli, he first applied semi-quantitative fluorescence microscopy and biochemical approaches to characterize aging-related molecular pathways that maintain heterochromatin structure and protect cells from DNA damage. Then, in 2009 Dr. Pegoraro designed and implemented an imaging based siRNA screen for the identification of novel cellular pathways involved in Protein Quality Control (PQC). In 2012 he joined the United States Army Medical Research Institute of Infectious Diseases (USAMRIID)/PerkinElmer in Frederick, MD as a Research Scientist. While at USAMRIID, Dr. Pegoraro developed high-content imaging assays and imaging analysis routines for the study of host-pathogen interactions, and for the identification of novel antiviral compounds by high-throughput screening of small molecule libraries. In 2014 Dr. Pegoraro was appointed Head of the High-Throughput Imaging Facility (HiTIF)/LRBGE/CCR.