Introduction

The Biosciences FACS facility consists of a Becton-Dickinson FACS Vantage SE flow sorter and Becton-Dickinson FACS Calibur and FACS Scan flow cytometers. The facility was established by the Life Sciences Division and the mission of the facility is to support biosciences research at LBNL by providing capabilities which individual labs might not otherwise have access to.

The specifics of the instruments are:

1) BD FACS VantageSE high-speed sorter with 405, 488, and 633nm lasers, 6 fluorescence and 2 scatter channels, sorting capability into 96-well plates, DiVA digital electronics, different size nozzle tips, closed flow system for biohazardous material, optional sample cooler, etc.

2) BD FACS Calibur flow analyzer with 488 and 633nm lasers, 4 fluorescence and 2 scatter channels, optional automated high-throughput sampling capability from 96-well plates.

3) BD FACScan flow analyzer with 488nm laser, 3 fluorescence and 2 scatter channels.

The facility is managed by flow cytometry experts from the UCSF Lab for Cell Analysis, Sarah Elmes and Bill Hyun. For training and most questions, contact Sarah by email. She is available in person at the facility most Tuesday afternoons. LBNL oversight of the facility is provided by Damir Sudar and Heidi Feiler.

Getting access

The facility is available to all LBNL Biosciences reseachers with valid LBNL employee or guest appointments and who have access to a valid LBNL project ID for the usage charges. The systems are located in the Berkeley West Biocenter (a.k.a. Potter St) in room 106. Users can arrange with Sarah Elmes to run their samples at the higher "with operator" charge rate on Tuesday afternoons. This is primarily meant for experiments on the complex high-speed sorter system. Contact Sarah to make arrangements. After completing training users may use the analyzers and after very extensive training the sorter. Training sessions can be scheduled on Tuesday afternoons by emailing Sarah. As part of the training users will be asked whether they completed the relevant EH&S training and whether the use of the FACS facility has been documented in the Biological Use Authorization (BUA) for the home lab of the user. Access to the FACS facility is prohibited until completion of instrument training, EH&S training, and BUA documentation. See EH&S information (below) for more detail on these issues.

Once trained, users can sign up for and use the systems for which they completed training. Sign up through the LBNL calendar system by reserving the resource "977-106 FACS Calibur" or "977-106 FACS Sorter". See: https://www.lbl.gov/wiki/bin/view/Main/CalendarFaq for all information about the LBNL Calendar system including how to get an account.

In order to log in on the computer that controls the FACS instrument, an account is needed on LBNL's Active Directory or Windows Domain system. (Note: This is different from the LDAP account that is used for email, LETS, etc.) Many LBNL users already have an account and can simply use that to log in. If not, visit: http://www.lbl.gov/IT/CIS/accounts.html to request a Windows Domain account. See: https://www.lbl.gov/wiki/bin/view/Main/ActiveDirectory for more information about Active Directory.

Prior to each use of an instrument in the FACS facility, users must complete an entry in the Run Log which is located in the binder next to the instrument. This will document and confirm that the user is familiar with and will follow all rules that govern the use of the FACS facility. Violation of these rules will result in disciplinary action including denial of access to the facility.

The facility will provide only sheath fluid and the supplies needed to maintain and calibrate the instruments. All other supplies must be brought by the users. This includes the standard 5ml test tubes, 40 micron strainers, transfer pipettes, gloves, etc. These supplies can be purchased from VWR. In the Resources section of this page (right column) is a list of the typical supplies and the ordering information.

EH&S 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 EH&S 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 EH&S training website for more information. The use of the FACS facility and the method of transport of samples between the home lab and the FACS facility must be documented in the BUA of the Principal Investigator. To facilitate this we have prepared an EH&S-approved insert that can be made part of the PI's BUA. If you have any questions about this insert, please contact Damir Sudar for information. In addition, every FACS user should be familiar with the FACS Standard Operating Procedure document. Ensuring that all EH&S 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 and will be kept for 1 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 we recommend the FlowJo program which we can make available for a reduced fee through our collaboration with UCSF. For an annual (July 1 through June 30) fee of $150 we can provide a license code 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 for further information.