CRB FACS Core
Center for Reproductive Biology
Fluorescent Activated Cell Sorting Core
FACS machines available to the WSU community on a fee-for-service basis

Fluorescent Activated Cell Sorting Core

The Fluorescent Activated Cell Sorting (FACS) core, directed by Dr. Jon Oatley, provides a fee-based service available to the WSU community. Investigators can isolate subsets of cells from a heterogonous population based on up to 8 different parameters.

The core contains two new state-of-the-art FACS machines manufactured by Sony Biotechnology that contain 4 lasers each.

Machines available for sorting

  • Sony SH800 – 4 lasers (405, 488, 561, 638), 2-way sort capabilities
  • Sony SY3200 – 4 lasers (405, 488, 561, 642), 4-way sort capabilities

Flow cytometry is a powerful tool for critically analyzing individual cells and other biological particles. Using this technology, expression of proteins on the surface of cells as well as simultaneous assessment of cell size and complexity can be investigated. A component of flow cytometry is fluorescent activated cell sorting (FACS), which allows for specific subsets of cells to be isolated in purity from a heterogeneous population. Moreover, isolation of single cells can be achieved using FACS allowing for critical examination of heterogeneity within diverse populations.

Contact

The cell sorting instruments are housed in the basement of the Veterinary and Biomedical Research Building (VBRB). Hours of operation are 8 a.m. to 5 p.m. Monday through Friday.

Initiation of service is made by contacting the FACS Core Service Manager Melissa Oatley. A pre-service meeting will be scheduled to discuss details of an individual investigator’s needs and technical considerations for setting up FACS experiments using the core’s specific machines. Service charges are established to cover costs of operating the machines and service contracts.

Sorting service is conducted by the service manager based on the investigator’s pre-defined parameters.

Service charges are established to cover costs of operating the machines and service contracts.

  • Appointments: If possible, appointments need to be made with a 1-week advance notice to ensure the availability of an instrument and sorting technician. Under special circumstances, an appointment can be made with at least a 48-hour advance notice. The clock for determining service fees starts at the agreed upon appointment time.

Baring machine or operator error, the PI will be billed for all sorts regardless of whether the desired outcome is achieved. The core cannot guarantee a desired result if samples have not been properly processed for sorting or controls to set the sorting parameters are lacking.

SY3200 
Standard rate (Center members, WSU, & federal)$119.00/hr
Outside WSU$161.00/hr
Minimum Charge: 1 hour
WSU out of area & outside WSU: 
8.7% administrative charge will be added
Cancelation policy: The minimum charge will be invoiced if an appointment is canceled with less than a 3-hour advance notice, 50% the minimum rate will be charged with less than a 24-hour notice.
SH800
Standard rate (Center members, WSU, & federal)$116.00/hr
Outside WSU$156.00/hr
Minimum Charge: 30 minutes
WSU out of area & outside WSU: 
8.7% administrative charge will be added
Cancelation Policy: The minimum charge will be invoiced if an appointment is canceled with less than a 1-hour advance notice, 50% the minimum rate will be charged with less than a 24-hour notice.

The CRB will review all rates and accounts as per WSU policies and procedures.

  • Controls are vital to a successful sort. Negative controls (e.g., cells incubated with normal isotype antibody or cells from wild-type litter mates when working with fluorescent reporters) are necessary for setting gates to collect the desired population.
  • Samples must be a single cell suspension in a sorting media that has less than 1% FBS and little to no phenol red. Reagents are available from commercial sources for disassociation of tissues with a propensity for cell clumping. Note cell viability should be assessed after generation of single cell suspension and prior to sorting.
  • Sample tubes:
    • SH800 – Samples can be processed in 0.5 mL, 1.5 mL, 5 mL, or 15 mL conical tubes
    • SY3200 – Samples can only be processed in 5 mL round bottom tubes
  • The ideal sample concentration will vary depending on the cell type. A good starting point is 4×106 cells per mL.
  • Collection tubes:
    • Collection tubes are supplied by the investigator and should contain at least 250 uL of collection buffer. If post sort assessment is desired, the buffer should contain no more than 1% FBS.
    • Tube sizes:
      • SH800 – 5 mL round bottom and 15 mL conical tubes
      • SY3200 – 5 mL round bottom and 15 or 50 mL conical tubes, and any size flat bottom well plate