Center for Molecular Microscopy (CMM) is an interdisciplinary research collaboratory, located primarily at the ATRF, specifically aimed at applying recent cutting-edge advances in the field of 3D electron microscopy to important problems within the CCR portfolio. The research program of CMM includes the development and application of novel methods for cellular imaging using correlative light and focused ion beam scanning electron microscopy, and molecular imaging using cryo-electron microscopy and cryo-electron tomography. The Genomics Technologies Lab (GTL) provides services to CCR investigators in Affymetrix microarrays, Whole Exome capture and sequencing, Nanostring gene expression analysis and NGS sequencing. Additional services have been provided in the genomics lab at lower levels of support. The Protein Expression Lab (PEL) provides dedicated support to CCR investigators for cloning, protein expression, and virus production. Cloning services includes the construction of expression plasmids as well as clones that influence and report protein expression (shRNA, microRNA, reporter plasmids). For microbial and eukaryotic expression, transient and stable expression of proteins in a variety of systems including, E.coli, yeast, insect, mammalian cells, or cell-free expression is provided. The lab provides parallel small scale screening in bacteria, yeast, insect, and mammalian cells to optimize protein quality using appropriate platforms to test multiple expression modalities side by side to determine for each target protein the most suitable method for purification. PEL also provides scaled-up production for tens of liters of culture and hundreds of milligrams of protein. Custom recombinant lentivirus and adenovirus production to study transgene or shRNA delivery for gene knockdowns in vitro and in vivo is also offered, and includes virus concentration and titration services. The Protein Chemistry Lab (PCL) provides dedicated support to CCR investigators for protein quantitation, identification and characterization, including protein interaction partners isolated from mammalian cells. Protein Identification is performed from SDS PAGE gels, co-immunoprecipitation and related methods. Qualitative and quantitative analyses using high resolution, accurate mass data for high confidence identifications and good quantitative reproducibility is provided using a variety of mass spectrometry techniques. The lab has capability to identify posttranslational protein modifications such as glycosylation, and performs N-terminal amino acid sequencing using protein sequencer. Biacore/SPR analysis to measure binding affinities is also available, as is measurement of metabolic flux in cells using the Seahorse XF24 Analyzer. The Optical Microscopy and Analysis Laboratory (OMAL) provides, develops and applies technologies for quantitatively understanding the molecular basis of cancer and HIV infection in the spatial-temporal context of the cell and tissue.The available technologies range from standard fluorescence microscopy to super-resolution and intra-vital two-photon imaging, and computational capabilities for visualization and analysis. A new Single Cell Analysis Facility was established at the ATRF and the Bethesda main campus. These technologies will give CCR-investigators the options for single cells studies, ranging from a few cells to hundreds of thousands of cells, and from single cell genomic assays such as RNA-seq to single cell protein quantification.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Scientific Cores Intramural Research (ZIC)
Project #
1ZICBC011596-06
Application #
10015080
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Budget End
Support Year
6
Fiscal Year
2019
Total Cost
Indirect Cost
Name
National Cancer Institute Division of Basic Sciences
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