Flow Cytometry Facility

UCSC aerial photo of monterey bay and campus
BC Cytoflex with computer visible on lab mench

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 TrainingUCSC UsersNon-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 SortsUCSC UsersNon-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.

FACS internal image courtesy of FHCS

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.

group of petri dishes sitting on lab bench with calculator in background

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