ZIC BC 011428 (ZIC) | |||
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Title | CCR, LGI, Flow Cytometry Core Facility | ||
Institution | NCI, Bethesda, MD | ||
Principal Investigator | Livak, Ferenc | NCI Program Director | N/A |
Cancer Activity | N/A | Division | CCR |
Funded Amount | $837,598 | Project Dates | 00/00/0000 - 00/00/0000 |
Fiscal Year | 2017 | 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 (29.0%) |
Brain (15.0%) Breast (15.0%) Colon/Rectum (10.0%) Leukemia (2.0%) Liver Cancer (15.0%) Lung (10.0%) Melanoma (5.0%) Nervous System (15.0%) Non Hodgkins Lymphoma (3.0%) Ovarian Cancer (5.0%) Pancreas (4.0%) Prostate (15.0%) |
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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, 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 older, two-laser, analog BD FACSCalibur cytometer. The two identical 18-color LSRFortessa SORP instruments ensure that despite heavy utilization users can perform the most demanding experiments with minimum wait time and maximum flexibility. In addition, the Core installed a new Sony SA3800 spectral flow analyzer during FY17. This instrument relies on the novel spectral flow cytometry technology which collects fluorescence data on the entire visible spectrum of 500-800nm range in real time for every single cell. An innovative software unmixes the spectral data to identify every each fluorescent reagent that is used in the experiment. The instrument utilizes a High Throughput Sampler (HTS) loading mechanism to enable investigators to analyze large number of samples either from FACS tubes of from 96 well plates. The core also operates four cell sorters: two BD FACSAria II instruments equipped with three lasers with the capacity to simultaneously identify 9-11 fluorochromes. One Beckman Coulter MoFlo Astrios EQ instrument, housed in a custom-designed Class II biosafety cabinet, is equipped with five lasers and capable of simultaneously detecting 20 fluorochromes. This instrument was upgraded with an additional, enhanced forward scatter photomultiplier tube that permits analysis and sorting of microparticles with sizes substantially below the typical resolution limit of other cell sorters. The BD FACSAria Fusion cell sorter is also housed in a custom-designed Class II type A2 biosafety cabinet and is equipped with five lasers and has the capacity to simultaneously identify 18 fluorochromes. This instrument has similar capabilities to the MoFlo Astrios and these two sorters have configurations essentially identical to the two high-end LSRFortessa SORP analytical flow cytometers. This conscious instrumentation planning has resulted in a core service where users can seamlessly transfer advanced, complex multi-parameter analytical panels to cell sorting without the need for extensive further optimization or compromising their experimental design. Further, the Class II biosafety cabinets, combined with the separate housing in a dedicated labo |