FACS Facility: Flow Cytometry/Sorting
Instruments in the FACS Facility include a Becton-Dickinson FACSAria high speed cell sorter, and a FACSCalibur flow cytometer. The mission of the facility is to support biosciences research at Berkeley Lab by providing capabilities which individual labs might not otherwise have access to. Others interested in using the facility please see below under “Getting Access.”
The facility is managed by David Acevedo along with backup support provided by flow cytometry experts from the UCSF Lab for Cell Analysis (LCA), Sarah Elmes and Bill Hyun. For training and most questions, contact David; he can be reached via phone at (510) 486-6139 and e-mail.
The facility is available to all Berkeley Lab reseachers with valid Berkeley Lab employee or affiliate appointments, and who have access to a valid Berkeley Lab project ID for the usage charges. Others interested in using the facility, please request access via the Science Exchange website or contact David Acevedo.
The facility is located at 717 Potter Street, Berkeley. Training is required before one can use the analyzers; more extensive training is required to use the sorter. Training sessions can be scheduled by emailing David Acevedo. Relevant Environment/Health/Safety (EHS) training also needs to be completed prior to access.
Once trained, users can sign up for and use the systems for which they completed training. Sign up through the Facility Online Manager (FOM) system, selecting “Lawrence Berkeley Laboratory Cytometry/Microscopy” facility. For instructions on the FOM system see this document; and refer to this document on how to add this scheduling tool to a Google or Outlook calendar.
In order to log in on the computer that controls the FACS instrument, an account is needed on Berkeley Lab’s Active Directory or Windows Domain system (Note: This is different from the LDAP account that is used for email, LETS, etc.). Many Berkeley Lab users already have an account and can simply use that to log in. If not, visit this website to request a Windows Domain account. See here for more information about Active Directory.
Read the section below for information about usage charges and supplies.
Usage Charges & Supplies
The charge for any instrument is $29.00 per hour. Also, if operator assistance is needed an additional fee of $120.00 per hour is charged (Fee schedule). For external users, additional charges will apply; please use the Science Exchange website for access to our facility and to request a quote. Users are charged via the Facility Online Manager (FOM) system. Please download and read the instructions for setting up an account.
The facility provides Heath fluid and cleaning solutions for each cytometer. All other supplies must be brought by the users, which includes slides, transfer pipettes, gloves. In the resources section below is a list of the typical supplies and the ordering information.
The FACS Facility is located in a BSL-2 lab; no food or drink is allowed. Each user must have completed and be current on the following EHS training: EHS0735/738 Bloodborne Pathogen training and annual refresher, EHS0745 Hepatitis B Medical Surveillance training, EHS0604 Hazardous Waste Generator training, and EHS0730 Biohazardous Waste training. Visit the Berkeley Lab training website for more information. The use of the FACS Facility and the method of transport of samples between the home lab and the the facility must be documented in the BUA of the Principal Investigator. If you have any questions, please contact David Acevedo for information. In addition, every FACS user should be familiar with the Standard Operating Procedure document. Ensuring that all EHS regulations are properly followed is the responsibility of the user and their Principal Investigator.
Data Storage and Analysis
The computers that control the FACS machines may temporarily be used to store acquisition data; these will be kept for one week after acquisition. Users are responsible for their own data and can copy it over the network to their lab’s server, copy it to a USB memory stick or portable USB hard drive, or burn it onto a CD or DVD. For flow data analysis the FlowJo program is recommended which can be made available for a reduced fee through a collaboration with UCSF. For an annual (July 1 through June 30) fee of $150 a license code is provided to run the program on the user’s own computer (PC or Mac). Simply download the FlowJo program, install it, and run it in evaluation mode. Once ready to purchase, contact Sarah Elmes for further information.
Four fluorescence channels and two scatter channels are available from the 488 nm laser (FSC, SSC, FL1, FL2, FL3, FL4).
Two fluorescence channels are available from the 405 nm laser (FL5, FL6).
Two fluorescence channels are available from the 635 nm laser (FL7, FL8).
Nozzle sizes from 50 to 400 micron are available depending on cell size.
Direct sorting into 5 ml and 15 ml tubes and direct cloning into 96-well plates are available.
FACSAria Quick Guide (Note: training is required)
The FACSCalibur is a bench-top cell analyzer, configured with two lasers:
1. 488 nm Blue-green Argon Ion laser;
2. 633 nm Red Helium Neon laser.
Three fluorescence channels and two scatter channels are available from the 488 nm laser (FSC, SSC, FL1, FL2, FL3).
One fluorescence channel is available from the 633 nm laser (FL4).
An optional High Throughput Sampler station is attached to allow direct sampling from 96-well and 384-well plates.
FACSCalibur user instructions (Note: training is required)
Facility Online Manager (FOM) system account set-up instructions
Facility Online Manager (FOM) system – how to add to Google or Outlook calendar?
FACSCalibur User Instructions (Note: training is required)
FACSAria Quick Guide (Note: training is required)
Google Drive Cytometry Folder, containing instrument manuals, user guides, etc.
FACS Standard Operating Procedure Document
If you have any questions about the FACS Facility, please contact David Acevedo.