Flow Cytometry Policies & Procedures | UC Davis Comprehensive Cancer Center

It is the user’s responsibility to read and understand the following information:

The UC Davis Flow Cytometry Shared Resource, or FCSR, provides analytic flow cytometry and cell sorting services at locations in Davis and Sacramento. To allow smooth operation and fair access to the equipment for all investigators and to maintain equipment in proper working order operational policies and procedures must be followed. Key operational policies are highlighted below. Please see additional information regarding other operational procedures in the following paragraphs.

Appointment policy:

1. Appointments for all instrument usage time must be scheduled.
2. Operator assistance is required for cell sorting.
3. Cell sorting appointments require a minimum of 24 hours advance notice.
4. Analytic cytometers: operator-assisted appointments require a minimum of 24 hours advance notice.
5. Approved, trained users may make appointments for self-operation same day.
6. Evening and weekend appointments are authorized for trained, self-operated use only. 

Appointment duration:

1. Cell sorters and the 5 laser Fortessa analytic cytometer: minimum one hour
2. All other analytic cytometers: minimum 30 minutes

Training sessions:

1. Training sessions are a minimum of two hours and will be charged at the assisted user rate. Users must supply a sample for training purposes.
2. The ability to independently operate the cytometers will be at the discretion of laboratory personnel.
3. Re-training: Self-operation privileges are earned and subject to approval by laboratory management. Re-training will be scheduled at the discretion of FCSR staff, and billed as assisted time.

Biohazard policy for analytic cytometers:

1. All samples run on bench-top analytic cytometers must be fixed.

Decontamination and fluidics washing:

1. Proper decontamination and cleaning of the fluidics tubing and flow cell must follow each use of the cytometer.
2. Post-acquisition washing and cleaning procedures are considered normal run time and will be billed as part of the total appointment time.
3. All users will wash the fluidics for a minimum of 15 minutes following each use of the cytometer; HTS users must add additional time for HTS cleaning.

Facility operating procedures and rules of conduct

The facility operating procedures described in this section ensure safe, efficient and equitable functioning. Any violation of these procedures may result in termination of access privileges.

Biohazards

Two instruments are available for use with biohazards: the Astrios EQ in Tupper Hall and the BD inFlux Cell Sorter in the Institute for Regenerative Cures. Any use of biohazards must be discussed with the laboratory manager before the application is approved. Facility personnel must be notified of potential biohazards prior to scheduling appointments and running samples. Users are required to dispose of all tubes and samples left after analysis by taking such materials back to their home laboratory in containers approved for safe transport.

All experiments must comply with UC Davis regulations for biosafety. All samples run on bench-top analyzers must be fixed. For sorting of biohazards, an amendment must be approved and filed with Environmental Health and Safety. Failure to meet facility biohazard requirements will lead to termination of access.

Operators and self-operation

Several analytic (non-sorting) cytometers are available in Davis and Sacramento for operation by trained personnel, and training is available for self-operation. Training is available by appointment only and will be performed using samples provided by the investigator. Laboratory personnel will determine when the investigator is able to self-operate. Keys to the facility are available for after-hours use.

Sorter self-operation is a major commitment and recommended only if the user will be sorting for a significant period of time. Self-operation of the BD FACS Aria II cell sorter in Sacramento may be allowed by the facility manager for approved users during hours when staff is available to assist should problems arise. Initial sorter training with the facility staff is followed by hands-on experience until sufficient skill is acquired to self-operate the Aria sorter.

Self-operators are expected to be capable of normal operations without the facility operator's assistance, including startup and shutdown, sorting, analysis, sterilization and maintenance (nozzle clogs, etc.). Sorter training and ability is required prior to self-operation authorization. Self-operators must negotiate with the facility operator for help when independently operating equipment. Extensive reliance on the facility operator assistance leads to full charges. Self-operators can sign up for 15 minutes of facility operator time and self-operate for the remaining time. Equipment abuse will lead to self-operator privilege curtailment until problems are clarified.

Signups

Operator-assisted runs are scheduled Monday through Friday, 10 a.m. to 4 p.m. Assisted use times can be negotiated after hours. Signups must be made on the Instrument Scheduling calendar. Users who sign up for assisted use are responsible for making sure that a staff member is available for assistance at that time. Valid users may schedule an instrument by selecting Instrument Scheduling from the menu.

Signup times are charged if not cancelled 24 hours before the run. If a cancellation occurs during the 24 hours prior to a run, the time becomes available for others and there is still a charge. If signup time is shifted or the facility operator makes an adjustment, the late cancellation charge may be waived.

Running time

A signup entitles the user to the specified time slot. A user with a reservation may insist that his or her samples be run within the specified period and may terminate the previous user's run. If equipment malfunctions or delays prevent usage, the user may request that charges be reduced accordingly. This strict schedule interpretation is intended to enable users and operators to meet obligations. Billing is for the actual time the instrument is in use (including the minimum described below). To discourage the reservation of instruments for unrealistically long periods, if more than an hour of a previously reserved slot remains unused, the user will be billed for the extra time.

Facility running time policy:

• Reservations are charged if not cancelled 24 hours in advance.
• Users must reserve 30 minutes minimum on all machines, except for the Fortessa 5 laser cytometer and the cell sorters, which require one hour.
• An extra charge will be incurred for runs that extend longer than the signup time.
• The facility operator must be notified if equipment is not usable.

Total running time can be adjusted when delays are due to:

• Previous users
• Equipment malfunction

Nozzle clog delays are caused by clumpy samples or excessive boost duration and are considered chargeable time. If the previous user left a clog or excessive time is needed to clear the flow-cell, running time adjustment will be made

Facility rates

Accounts are billed monthly through the Department of Medical Pathology. Billing questions should be directed to Bridget McLaughlin. A DaFis recharge number is required for facility use.

Data archival

Facility users are responsible for all data archival.

Problems

Users should report problems at the facility immediately to:

• Report instrumentation problems to Bridget McLaughlin.
• Director Dr. Barbara Shacklett is the final arbiter for problems that arise at the facility.