NATIONAL CANCER INSTITUTE - CANCER.GOV

Contact Information

Primary Contact

Gianluca Pegoraro
Facility Head

Location

41 Medlars Drive
Building 41 Room C508
Bethesda, MD 20892

Additional Contacts

Adib Keikhosravi
Image Analysis Scientist
Isabel Quintanilla Leo
Assay Development Scientist
Laurent Ozbun
Automation Specialist
MD Abdul Kader Sagar
Image Analysis Scientist

Overview

The NCI High-Throughput Imaging Facility (HiTIF) works in a collaborative fashion with NCI/NIH Investigators by providing them with the necessary expertise, instrumentation, and software to develop and execute advanced High-Throughput Imaging (HTI) assays. These can be paired to screen libraries of RNAi or CRISPR/Cas9 reagents to discover and characterize novel cellular pathways by functional genomics. In addition to functional genomics screens, HiTIF also develops HTI assays to extract and quantify single-cell information about the molecular mechanisms underlying rare or heterogeneous biological events in cellular populations.

To initiate a project with the HiTIF, the investigator first needs to request a consultation with the Facility Head via the HiTIF iLab Webpage.

Established Technologies

HiTIF Core provides infrastructure and expertise to NCI intramural scientists for:

  • Fully automated, high-throughput fluorescence and bright-field microscopy (Yokogawa CV7000, Yokogawa CV8000)
  • High-Content Analysis (HCA) to measure up to hundreds of cellular features from microscopy images
  • Assistance with miniaturization of cell-based assays in 96-well and 384-well microplates
  • Robotic sample preparation, reagent addition, and plate washing
  • Characterization of cellular mechanistic pathways using RNAi- and CRISPR/Cas9 Knock-out screens
  • High-throughput live-cell fluorescence microscopy
  • Automated tracking and image processing of subcellular objects in live cells
  • Automated RNA in situ hybridization methods for single molecule detection of endogenous genomic DNA loci, or mRNA transcripts

Developing Technologies

  • Third-generation automated microscopy instrument optimization/development
  • 3D HTI assays
  • Deep Learning methods for cellular segmentation and classification of cellular objects

Major Instrumentation

Yokogawa CV8000 High-throughput spinning disk confocal microscope

Yokogawa CV7000 microscope with integrated robotic arm and plate hotels for automated plate feeding

Auxiliary Instrumentation:

PerkineElmer Janus Automated Liquid Handler

Beckman ECHO525 Acoustic Liquid Handler

Biotek EL406 Plate washer/Dispenser

BlueCatBio Bluewasher/Dispenser

PerkinElmer Columbus image storage and analysis server

User Guidelines

The HiTIF runs projects on a collaborative basis with NIH investigators. Precedence will be given to CCR/NCI projects. Requests for information, discussion of possible collaborations and for training, and access to the HiTIF equipment should be directed to the Facility Head.  Investigators are expected to cover the cost of reagents and consumables.

Keywords

CRISPRHCSHTSassay developmentImaging and Microscopyfunctional genomics screeninghigh content screeninghigh throughputhigh throughput screeninglibrary managementmicroscopynci-coresiRNAsiRNA screeningSingle Cell Analysis