Flow Cytometry Grants and Publications

Flow Cytometry Grant Descriptions

 

Short Grant Description

The Flow Cytometry shared resource provides access to state-of-the-art cytometry instrumentation, including 13 cell analyzers (8 conventional and 5 spectral systems) and 1 Helios CyTOF mass cytometer, located at two sites: the E level of the Thomas building and the first floor of the Steam Plant building. The analyzers offer detection capabilities from basic 3-laser high-throughput systems to high-parameter systems for deep immune profiling with >50 markers on the single cell level.

Researchers have access to 8 cell sorters, including 1 Miltenyi Tyto, 3 Sony MA900, 3 BD (Becton Dickinson) FACSymphony S6, and 1 BD FACSDiscover S8 spectral imaging cell sorter. The experienced technical staff provide consultation, training, assisted analysis, and cell sorting services to support the Fred Hutch/University of Washington/Seattle Children’s Cancer Consortium members.

 

Long Grant Description

Overview: The Flow Cytometry shared resource at Fred Hutchinson Cancer Center operates laboratories in two locations on campus: the E level of the Thomas building and the first floor of the Steam Plant building with a total of 21 instruments available to Fred Hutch/University of Washington/Seattle Children’s Cancer Consortium members. The core is supported by a staff of four Cytometry Specialists, two Cytometry Leads and one Director with a combined 50+ years of experience.

The core houses 13 cell analyzers, with a range of capabilities, including conventional to spectral systems with high-throughput capabilities for deep immune profiling of >50 markers on the single cell level. Researchers have access to 8 cell sorters; each is housed in a separate room within an approved biosafety cabinet with additional aerosol management systems allowing live cell sorting of BSL2 and BSL2+ specimens.

The instruments and their capabilities are detailed below.

Cell analyzers: There are 8 conventional and 5 full spectrum systems available, each with tube and plate loader options. The conventional analyzers include; five 5-laser BD (Becton Dickinson) FACSymphony A5’s, two are spectrally enabled (SE) systems which expands panel capability up to 32 colors, one 5-laser BD LSRFortessa X50, three 4-laser BD FACSCelesta’s, and a BD FACSymphony A3. The three spectral analyzers; a 7-laser Sony ID7000, and two 5-laser Cytek Auroras are capable of supporting panels with 40 or more colors. Additionally, there is one CyTOF Helios mass cytometer capable of supporting panels of more than 40 markers with no spreading error due to fluorescent compensation increasing sample resolution for deep immune profiling.

Cell sorters: The core houses a variety of cell sorters at the two locations to meet the diverse needs of researchers. The cell sorters are capable of a range of pressures and nozzle sizes from 70um - 130um to accommodate most cell types. Each sorter is monitored daily for optical performance and system sterility.

The flow core cell sorters are as follows:

One 3-laser Miltenyi Tyto fully enclosed cartridge-based sorter, with 8 color detections, small volume sample collection and gentle mechanical sorting which lowers the impact and activation of sensitive cell types.

Three 4-laser Sony MA900s with easy-to-learn software for panels up to 12 color cell sorting into both tubes and multi-well formats.

Three 5-laser BD FACSymphony S6s, 2 are SE all are equipped with UV lasers for up to 32 color detection and capable of 6-way and multi-well single cell sorting.

One 5-laser BD FACSDiscover S8 spectral imaging cell sorter capable of supporting >50 color panels with six real-time imaging parameters. The FACSDiscover S8 is the first-of-its-kind imaging cell sorter allowing researchers to observe morphological and spatial characteristics on the single cell level. The technology allows sorting based on phenotypical fluorescence and additionally on visual characteristics of cells with confirmation of doublets for increased precision of cell sorting.

All equipment is connected to UPS devices to ensure stability of electrical components, and all supporting equipment is regularly maintained by flow core staff and Fred Hutch Facilities & Engineering.

Services: The experienced technical staff provide users with consultation on panel design, data analysis and interpretation. Researchers have access to training and educational resources both online and in person. One-on-one sessions can be arranged for assisted sorting, hands-on training for independent usage, and after-hours access.