The facility provides access to high-speed cell sorting, analytical flow and imaging flow cytometry on the SBP campus in La Jolla. Trained SBP investigators have 24-hr/day access to a variety of analytical flow cytometers available for independent use. Trained external users have access during business hours.
Core staff provide technical expertise, hardware and software training, operate the facility\'s cell sorters and are available to assist with analysis experiments for those who prefer to have their samples run by an expert cytometrist.
Scientists planning a flow cytometry experiment are encouraged to consult facility staff for assistance with protocols, fluorochrome selection or other aspects of experiment design.
Other Facility Information
- Single-cell (clone) sorting into 96-well plates
- Simultaneous sorting of up to 4 populations
- Analytical flow cytometry: do-it-yourself 24hr/day, or assisted by appointment
- Imagestream Imaging Flow Cytometer
- Hardware and software training
- Consultation
- Experiment design
- Data analysis and interpretation
Facility Policies
Services are offerred outside of Sanford Burnham Prebys Medical Discovery Institute
Consulting is offerred outside of Sanford Burnham Prebys Medical Discovery Institute
Aria has four lasers, 405, 488, 561 and 633 nm, and can detect light in sixteen photomultiplier tubes. Its primary function is to examine complex populations of cells and yield pure populations of cells. The Aria can sort lymphocytes at a rate of approximately 15,000 events per second. For larger cells, the rate is much slower in order to optimize the passage of cells through the instrument. Cells from single populations can be sorted into 96-well plates. Alternatively, the Aria can sort into two 15-ml tubes or four 5-ml tubes. The Aria can purify samples that are simply positive and negative for a single fluorophore or as complex as nine-color samples with intricate gating schemes. The BD FACSAria IIu cell sorter is a high speed benchtop digital flow cytometer equipped with a sensitive, fixed-alignment cuvette flow cell. It has three spatially separated lasers - 405 nm, 488 nm and 633 nm. Its primary function is to analyze complex populations of cells and yield pure populations of cells meeting defined criteria. The sorter can purify samples that are simply positive and negative for a single fluorophore or as complex as multi-color samples with complex gating strategies. Sorting can be performed into two or four tubes with speeds up to 25,000 events/second. The system sorts by incorporating cells from the sample tube into a stream of sterile PBS. The stream is interrogated by the lasers at the flow cell and the system electronics keeps track of each cell as they pass through the laser and determines specific cells that meet the sort criteria. A transducer vibrates the stream and induces droplet formation, with cells in the stream being incorporated into the droplets. If a cell meets the sorting criteria and is in the last drop before the break off, the instrument will charge that drop. The charged droplet is then deflected into the proper collection tube by the charge plates. Different cell types can be sorted with the use of 70 ?m, 85 ?m and 100 ?m nozzles. If you need a yellow-green (561 nm) laser, check the BD FACSAria III instead. [Product Link]
BD LSRFortessa cell analyzer offers the ultimate in choice for flow cytometry, providing power, performance, and consistency. Designed to be and expandable, the BD LSRFortessa has the flexibility to support the expanding needs of multicolor flow cytometry assays. The BD LSRFortessa system can be configured with up to 7 lasers*,blue, red, violet, UV and yellow-green. The instrument can accommodate the detection of up to 18 colors simultaneously with a defined set of optical filters that meet or exceed the majority of today's assay requirements. BD FACSDiva software controls the efficient setup, acquisition, and analysis of flow cytometry data from the BD LSRFortessa workstation. The software is common across BD FACS instrument families, including the BD FACSCanto cell analyzer and BD FACSAria cell sorter systems. [Product Link]
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Citation:
Sanford Burnham Prebys Medical Discovery Institute Flow Cytometry Facility (RRID:SCR_014854)
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