City of Hope’s Analytical Cytometry Core (ACC) Facility provides researchers with high-quality flow cytometry instrumentation, as well as expertise in analyzing and/or sorting sample populations of interest via interpretation of their physical, fluorescent and/or light-scattering properties. The facilities and their services are available to both City of Hope and non-City of Hope researchers.
The ACC Facility has three components: the Flow Cytometry Facility (FCF), the Laser Scanning Facility and the Molecular Imaging Facility (MIF).
As an added convenience, the ACC supports several off-line workstations throughout the campus where investigators can analyze their ACC-derived data. FCF software includes Flowjo, Modfit and Summit. MIF software includes ImageQuant, PDQuest (Bio-Rad) or Odyssey.
The Flow Cytometry Facility has three instruments. Both the CyAn and FACScalibur are available for self runs. The MoFlo is operated by core staff only:
- The MoFlo MLS Cytometer (Dako) uses four available lasers to provide excitation from 351-633nm and is set up to collect up to nine fluorescent parameters. This instrument is a high-speed sorter capable of sorting four populations at a time. Plate sorting is also available for generating single cell clones.
- The CyAn ADP 9color (Dako) has three excitation sources (405nm, 488nm and 635nm) and is capable of collecting up to nine fluorescent parameters. It also has an automated 96/384 well plate delivery system for high-throughput screening applications (HyperCyt—IntelliCyt Corporation).
- The FACScalibur (BD) has two excitation sources (488nm and 635nm) and is capable of collecting four fluorescent parameters.
Laser Scanning Cytometry Facility
Our Laser Scanning Cytometry Facility uses a microscope and laser-based cytofluorometer for rapid measurement of fluorescent parameters of cells and tissues. The facility has the following equipment:
- iCys laser scanning cytometer from CompuCyte, which has three excitation sources (405nm, 488nm and 633nm) and eight possible filter sets. This instrument provides both quantitative and qualitative data from a wide variety of samples, including embedded tissue sections, single cell preparations and cell cultures. The technology allows for automated analysis of samples stained with fluorescent dyes coupled to antibodies or having specificity for unique cellular components (DNA, membranes, etc.). Along with fluorescent images, transmitted light images may also be obtained. The instrument collects positional and quantitative data for each measured event and provides coordinated values across multiple parameters.
Molecular Imaging Facility
This facility contains instruments utilizing lasers to scan gels, membranes (e.g., nylon, nitrocellulose), microtiter plates or thin-layer chromatography plates. The molecules of interest (DNA, RNA or proteins) are labeled with radioactive or fluorescent tags. There are two types of instruments available:
- The Typhoon 9410s (General Electric) has four excitation sources (457 nm, 488 nm, 532 nm and 632.8 nm). The Typhoon can also be used to acquire radioactive labeled data with a special phosphor screen, which is used to capture quantitative information that is then scanned on the Typhoon. This facility contains two Typhoon scanners.
- Odyssey Infrared Imager (Li-Cor) has two excitation sources (680 nm and 780 nm).
Last Updated: 06/30/08