The IBSC Flow Cytometry Facility offers multicolor cell analysis by a Beckman Coulter Cytoflex and single-cell sorting by a BD Biosciences FACS Aria cell sorter. The Facility is located on the 4th floor of the Biomedical Sciences Building and is available to any interested researcher.
We offer fully assisted cell sorting and analysis by appointment. The facility manager will work with new users on experimental design and protocol development. Users may become self-operators if they expect to use the Cytometry Facility regularly.
| Assisted Sorts and Instrument Training | UCSC Users | Non-profit Users not UCSC affiliated | For-profit Users not UCSC affiliated |
|---|---|---|---|
| FACS Aria | $95.30 | $308 | $570.63 |
| LSRII | $81.98 | $229.90 | $419.38 |
| CytoFlex | $72.71 | – | $419.38 |
| Unassisted Sorts | UCSC Users | Non-profit Users not UCSC affiliated | For-profit Users not UCSC affiliated |
|---|---|---|---|
| FACS Aria | $46.59 | $157.30 | $302.50 |
| LSRII | $33.27 | $79.20 | $151.25 |
| CytoFlex | $24.00 | – | $151.25 |
| Cytoflex LX (6-L UV) | |||||||
|---|---|---|---|---|---|---|---|
| Laserline | Filter | Fluorochromes | |||||
| UV 355nm – 20mW |
405/30 BP | Indol | BUV395 | ||||
| 450/45 BP | DAPI | Hoechst Blue | |||||
| 675/30 BP | BUV661 | Hoechst Red | |||||
| VIOLET 405nm – 80mW |
450/45 BP | Pacific Blue | V450 | Efluor450 | BV421 | DAPI | Supernova v428 |
| 525/40 BP | Krome Orange | V500 | BV510 | ||||
| 610/20 BP | Violet 610 | BV605 | Supernova v605 | ||||
| 660/20 BP | Violet 660 | BV650 | |||||
| 763/43 BP | Violet 780 | BV785 | Supernova v786 | ||||
| BLUE 488nm – 50mW |
525/40 BP | FITC | Alexa488 | CFSE | GFP | ||
| 610/20 BP | ECD | PE-Texas Red | PE-CF594 | PI | |||
| 690/50 BP | PC5.5 | PC5 | PerCP | PerCP-CY5.5 | |||
| YELLOW-GREEN 561nm – 30mW |
585/42 BP | PE | PI | DSRed | |||
| 610/20 BP | ECD | PE-Texas Red | PE-CF594 | PI | |||
| 675/30 BP | PC5.5 | PC5 | |||||
| 710/50 BP | PE-AF680 | ||||||
| 763/43 BP | PC7 | PE-CY7 | |||||
| RED 638nm – 50mW |
660/20 BP | APC | Alexa Fluor 647 | ||||
| 712/25 BP | APC-Alexa700 | Alexa Fluor 700 | |||||
| 763/43 BP | APC-Alexa750 | APC-H7 | APC-CY7 | APC-Efluor780 | |||
| INFRARED 808nm – 60mW |
840/20 BP | Alexa Fluor-790 | |||||
| 885/40 BP | PF840 (viability) | Ifluor850 (AAT Bioquest: 1408) | |||||
| BD FACS Aria Iiu | |||||||
| Laserline | Filter | Fluorochromes | |||||
| UV 355nm – 20mW |
379/28 | BUV 395 | |||||
| 740/35 | BUV 737 | VIOLET 405nm – 50mW |
450/50 | BV-421 | |||
| 525/50 | BV-510 | ||||||
| 605/40 | BV-605 | ||||||
| 670/30 | BV-650 | ||||||
| 710/50 | BV-711 | ||||||
| 780/60 | BV-786 | ||||||
| BLUE 488nm – 100mW |
488/10 | SSC | |||||
| 530/30 | FITC | ||||||
| 695/40 | PerCP-Cy5.5 | ||||||
| YELLOW-GREEN 561nm – 100mW |
586/15 | PE | |||||
| 610/20 | PE-CF594 | mCherry | |||||
| 660/20 | PE-Cy5 | ||||||
| 695/40 | PE-Cy5.5 | ||||||
| 780/60 | PE-Cy7 | ||||||
| RED 640nm – 40mW |
670/30 | APC | |||||
| 730/45 | Alexa 700 | ||||||
| 780/60 | APC-Cy7 | ||||||
Access
The Sinsheimer TC facility is open to any UCSC researcher. The Biomedical Sciences TC facility gives priority to IBSC investigators culturing stem cells. Flow Cytometry facilities are open to any UCSC or outside research projects.
Biohazard Procedures (BSL2)
All users must have an approved BUA on file and must fill out the UCSC IBSC and SSCF safety questionnaire (PDF) before doing any project in the facility.
Training
All new cytometry and tissue culture users are expected to have taken the “Biological Agents in Shared Facilities Awareness Training” class, which is available through our campus Learning Center (login and “Find a Course” to search).
Please contact the IBSC facility manager to discuss your project and arrange facility/instrument training.
Scheduling
Cell Culture Facilities: Access to hoods in the centers is on a first-come use policy. If you will require a hood for an extended period of time on a given day, please make arrangements with the IBSC facility manager.
Cytometry Instruments: Trained users, please use the online scheduling system to reserve equipment.
Acknowledgements in publications
In all publications of research conducted in the Sinsheimer cell culture lab, please acknowledge the CIRM Shared Stem Cell Facility grant to UCSC (CL1-00506) and the facility number (RRID:SCR_021353).
In all publications of research conducted on the Biomed 4th floor (including the cell culture and virus room), in the vivarium, or using the LSR II cell analyzer/FACS Aria cell sorter, please acknowledge the CIRM Major Facility Award to UCSC (FA1-00617) and the appropriate facility number for the cell culture facility (RRID:SCR_021353) and/or the cytometry facility (RRID:SCR_021149).
Reporting Problems, Suggestions, Questions
To report problems or make suggestions concerning the Sinsheimer and Biomedmedical stem cell facilities, please email the IBSC Facility Manager. Please specify the issue in your email.
New Users
New users need to turn in the IBSC safety questionnaire, and schedule a time with the facilities manager to discuss their project and experimental setup.
Cell sorting (FACSAria): New Users
Those users who do not anticipate routine work with the cytometer will perform all their sorts with the assistance of the facility manager, who will run the instrument with the provided samples. Users who expect to conduct frequent cytometry work (twice weekly or more for extended time periods) may request training to become a self-operator.
Cell analysis (Cytoflex): New Users
New users will be trained by Bari Nazario to use the Cytoflex.
Self-Operator
Cell sorting (FACSAria): Self-Operator
Users who wish to become self-operators undergo extensive training with Bari Nazario. After certification, self-operators are then provided with an account and login privileges, a key card for physical access to the room, and permission to operate the instrument without supervision.
Cell analysis (Cytoflex): Self-Operator
After training, users will be provided with a key card for physical access to the room, and permission to operate the instrument without supervision.
Operator Support
The Flow Cytometry facility is open 24/7 for APPROVED self-operator Aria cell sorting users, and TRAINED Cytoflex cell analyzing users. Keys and/or Card Key is REQUIRED at ALL TIMES to access the building and laboratory. Please contact the facility manager for a card key.
We offer operator support, by appointment, for people who need to schedule their FACS and Cytoflex times outside the normal staff hours. We cannot guarantee that every request can be accommodated, particularly on short notice. To request an operator appointment, please contact the IBSC facility manager. Include information about the instrument and times that would work for you, and include a good telephone contact number.
Funding/Grant Acknowledgement
This core facility is supported by the California Institute for Regenerative Medicine (CIRM) and the California Institute for Quantitative Biosciences (QB3). All publications, posters, presentations, etc. that utilize the Flow Cytometry Facility must acknowledge RRID SCR_021149.
Sinsheimer Cell Culture Lab and FACSAria
Publications of research conducted in the Sinsheimer cell culture lab, and research that utilized the FACSAria, please acknowledge the CIRM Shared Stem Cell Facility grant to UCSC (CL1-00506).
Cytoflex, Vivarium, and Biomed 4th Floor
Publications of research conducted on the Biomed 4th floor (including the cell culture and virus room); in the vivarium; or using the Cytoflex, please acknowledge the CIRM Major Facility Award to UCSC (FA1-00617).
Contact
Bari Holm Nazario, IBSC/SSCF Facility Manager
Email: bnazario@ucsc.edu
Phone: (831) 459-3980
Office: 450 Biomedical Sciences