Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research

Cell Analysis Core

Thanks to a grant from the Gordon and Betty Moore Foundation and another from the California Institute for Regenerative Medicine, The Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research at UCSF was able to acquire two FACS Aria II high-speed digital sorters and one FACS LSRII cell analyzer from Becton Dickinson in 2008-2009. The equipment is located in the Regeneration Medicine Building in Pod B, Room RMB-911E, and is managed by Michael Kissner.

Equipment

  • FACS LSRII from Becton Dickinson is designed for advanced analytical research applications using digital signal processing and flow cell optics, similar to the Aria systems. The LSRII is equipped with the latest technical capabilities - high-speed electronics, multiple high power* laser excitations across a wide spectral range (405nm, 488nm, 561nm and 640nm) and twenty photodiode/photomultiplier tube detectors for light scatter and fluorescence measurements.

  • FACS Aria II is a high-speed digital sorter that incorporates a fixed-alignment cuvette flow cell. Fiber optics direct and focus four high power* lasers (488nm, 640nm, 405nm, and 561nm or 355nm), depending on which system, onto the alignment prisms, and then focused on the cuvette flow cell. The flow cell allows for improved light excitation and collection optics. The collection optics consist of octagon- and trigon- shaped collection devices, and a total of up to 20 independent fluoroscence or light scatter signals can be acquired at one one time on a single-cell basis, followed by purification of cell subsets by sorting up to four particle populations into tubes simultaneously, or one population onto tissue culture or PCR plates. The instrument is equipped with four nozzle sizes for different cell types: 70 μm, 85 μm, 100 μm, and 130 μm.

*High power lasers are identical between the 3 systems: 60 mW 355 nm (UV), 100 mW 405nm (violet), 100 mW 488 (blue), 50 mW 561 nm (yellow/green), 40 mW 640 nm (red)

Getting Started

To establish an account, new users must complete the online OCF Calendar Account Creation Form. Once the form is received, we can set up an account for online instrument reservation in the OCF calendar. The Broad Center Cell Analysis and Sorting facility is primarily a user-operated facility. Therefore, all new users are required to either complete a training orientation with our staff, or, if the user has had previous experience, be assessed for proficiency in operation of the instruments. Additionally, Bill Hyun will continue to be available to help with advanced experiment design, Flowjo training, data analysis and optimal presentation of results. Users who need assistance running a sort with LCA will be required to book time through Core Admin at the LCA rate, as well as book time on the instruments through OCF. Please contact Bill (hyuncc@stemcell.ucsf.edu, 476-2632) for operator assisted cytometer experiments or sorts.

Scheduling Training and Use

First-time users should contact Michael Kissner at flowcoremanager@stemcell.ucsf.edu to arrange for training. Established users may view availability and schedule time on equipment through the OCF scheduler (http://irmimaging.ucsf.edu/login.php). Please include your phone number when scheduling training and use. Additionally, please include the type of sample you will be sorting or analyzing in your reservation.  Important: Please be aware that OCF is only accessible through the UCSF network. A VPN is required if you wish to access OCF from an off-campus location.

Instrument Rates

(per hour)

BD LSR II

Self Operation: $31.19

With-operator/Instrument training: $81.54

BD FACSAria II

Self operation: $45.17

With-operator/Instrument training: $95.52

 

Flow Cytometry Resouces

Please click here to access documents and guides relevant to flow cytometry instrumentation and applications.