Flow Cytometry Core
Flow cytometry is widely used in areas of research that require analysis or isolation of cells from suspension. This technology makes use of fluorescent probes targeted to specific cell-associated molecules to characterize the diversity and function of complex cell populations. In addition to analysis, fluorescence-activated cell sorters can individually identify and isolate live cells with defined phenotype that can later be expanded and/or further studied.
In addition to providing flow cytometry–related services and consultation, the core seeks to make the newest advances in flow cytometry technology available to the research community. With institutional support, the core sponsors and organizes technology seminars and flow cytometry–related basic courses for Cedars-Sinai users. The core is under the direction of Gislaine Martins, PhD, who is a member of the F. Widjaja Foundation Inflammatory Bowel and Immunobiology Research Institute and an assistant professor at the Department of Medicine and the Department of Biomedical Sciences Research, Division of Immunology. Research in the Martins Laboratory focuses on T lymphocyte function in health and disease.
New User Information
New users must have a registered iLab account and receive training from core personnel before using core equipment. Only after creating an iLab account and receiving training will new users be allowed to sign up on the online calendar and use Flow Cytometry Core equipment.
Sorter machines are only operated by Flow Cytometry Core staff. All sorter users must sign up in advance on the online calendar, follow the guidelines for preparing samples for sorting and complete the Pre-Sort Form within iLab appointment.
The Flow Cytometry Core staff is available for consultation on fluorochrome choice/sorting strategy for users who need help planning their cell sorting experiments.
When planning a flow cytometry experiment:
- Confirm with which instrument configuration the fluorescent labels and their combinations will be viewed.
- Read the sample preparation protocol.
- Schedule an appointment in the online calendar.
- Bring filtered samples to the core lab to be analyzed or sorted (if sorting, don’t forget collection tubes or plates.).
All scheduling is done through the online calendar.
- Analyzers are available for use by trained users 24/7. The last user of the day is responsible for turning off the machine and computer, even if they cancel their appointment.
- Sorting and assisted analysis take place Monday through Friday during regular working hours, with rare (experimentally justified) exceptions for after-hours sorting needs, which are evaluated and approved on a case-by-case basis. Pre-Sort Forms should be received a minimum of 24 hours prior to the scheduled sort time.
- Approximately 1 x 10^6 cells per mL in PBS containing 1% BSA. No phenol red should be used. Minimum sample volume recommended is 500 uL.
- Approximately 20-25 x 10^6 cells per mL resuspended in PBS containing 1% BSA. No phenol red should be used. Minimum sample volume is 500 uL.
- For single cell sorting into 96-well plates, cell concentration should be 1-5 x10^6 cells per mL.
Samples should be filtered prior to sorting. For intense cell aggregation, use DNAse, EDTA(1 mM) or PBS w/o Ca++ or Mg++ containing 0.1% BSA or FCS.
Sample Tubes: 12x75mm.
Collection Tubes:12x75mm(if sorting 3 or 4 populations) or 15mL (if sorting up to 2 populations). Collection tubes should contain an appropriate volume of media supplemented with 40% FCS.
The Cedars-Sinai Flow Cytometry Core is equipped with state-of-art equipment for cell analysis and sorting.
Specialized staff is available Monday through Friday to assist users in analysis and to perform cell sorting. The core is open for independent acquisition by trained users 24 hours a day, seven days a week.
The Flow Cytometry Core is now open for business, with new safety guidelines for new users, analyzer usage and sorting service usage.
Have Questions or Need Help?
Contact us if you have questions or would like to learn more about the Flow Cytometry Core at Cedars-Sinai.
Cedars-Sinai Flow Cytometry Core
8700 Beverly Blvd.
Davis Building, Room 4029
Los Angeles, CA 90048