ZIC BC 011428 (ZIC) | |||
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Title | CCR, LGI, Flow Cytometry Core | ||
Institution | NCI, Bethesda, MD | ||
Principal Investigator | Livak, Ferenc | NCI Program Director | N/A |
Cancer Activity | N/A | Division | CCR |
Funded Amount | $708,957 | Project Dates | null - null |
Fiscal Year | 2018 | Project Type | Intramural |
Research Topics w/ Percent Relevance | Cancer Types w/ Percent Relevance | ||
Ataxia Telangiectasia (5.0%) Bioengineering (10.0%) Cancer (100.0%) Digestive Diseases (31.0%) |
Brain (10.0%) Breast (18.0%) Colon/Rectum (12.0%) Leukemia (2.0%) Liver Cancer (15.0%) Lung (10.0%) Melanoma (5.0%) Nervous System (10.0%) Non Hodgkins Lymphoma (5.0%) Ovarian Cancer (6.0%) Pancreas (4.0%) Prostate (12.0%) |
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Research Type | |||
Resources & Infrastructure Related to Biology Technology Development and/or Marker Discovery |
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Abstract | |||
The mission of the CCR LGI Flow Cytometry Core in Building 37 (FC37) is to offer up-to-date instrumentation and technical expertise to CCR investigators to assist cancer research. The core provides full-scale, state-of-the-art flow cytometry services including analytical sample acquisition, data analysis, fluorescent cell sorting and experimental planning and consultation. The LGI FC37 also provides training on analytical instrumentation and cell sorters to permit investigators to independently acquire samples and perform data analysis. The core is committed to the dissemination of novel flow cytometry-based technologies through continuous education of investigators, sponsoring application seminars and participating in the newly developed NCI-wide flow cytometry training course with active contributions from director Ferenc Livak and two staff members, Karen W. Wolcott and Caiyi (Cherry) Li. In addition, the LGI FC37 strives to become a leader of excellence in flow cytometry applications by constantly seeking opportunities to enhance and broaden its technological platform in support of cutting edge cancer research. The LGI FC37 provides instrumentation for flow cytometry sample acquisition, data analysis, high-throughput sample acquisition, imaging flow cytometry, spectral flow cytometry and high-speed and single-cell sorting. The core is equipped with five analytical instruments: two high end BD LSRFortessa Special Order Research Product (SORP1 and 2) cytometers with identical configuration, equipped with five lasers and the capacity to simultaneously identify 18 fluorochromes. One of these instruments, LSRFortessa SORP1 is also equipped with a high throughput sampler (HTS) attachment that accommodates 96-well plates to further increase the capacity of the instrument. The core also has one three-laser, digital BD FACSCanto II cytometer and one Sony SA3800 spectral analyzer. In FY18 the Core removed its oldest, two-laser, analog BD FACSCalibur cytometer from service. In addition, in FY18 the Core added an entirely new technology, imaging flow cytometry to its portfolio (see below). The two identical 18-color LSRFortessa SORP instruments ensure maximum utilization and flexibility for our users. Nonetheless, rapidly increasing usage in FY18 prompted the Director of the Core to apply for funds to purchase a new, high-end cell analyzer. After a successful RRS application, the core is scheduled to receive and install a new 27-color BD FACSymphony analyzer in FY19, which will offer the highest analytical capabilities currently possible in fluorescent cell analysis. The increased use of analytical flow cytometry is accompanied by increasing demand of professional support for advanced panel design and data analysis, which lead to the active involvement of the Core to a soon-to-be published work with Michael Bustin's lab [LM]. In FY18 the Core installed a new ImageStream MarkII imaging flow cytometer (MilliporSigma). This instrument represents an entirely novel technology previously not available to CCR investigators at NCI. It records high quality images of every cell in flow and combines the high throughput of flow cytometry with the high information content of microscopic details. It expands the analytical capabilities of traditional flow cytometry by adding hundreds of morphological features to the fluorescent intensity measurements and allows unprecedented depth of quantitative, statistical analysis based on these fluorescent and morphological parameters. Several CCR labs have already begun to use this novel technology to study nuclear translocation of transcription factors (Ashwell and Samelson labs [LICB] and Annuziata lab [WMB]), quantification of nuclear DNA damage foci (Nussenzweig lab [LGI]), enumeration of circulating tumor cells (Wang lab [LMGC] and Hunter lab [LCBG]) and for the characterization of inflammatory responses (Trinchieri lab [CIP]) and microparticles and exosomes (Roberts lab [LP]). The core completed 698 sort |