Flow Cytometry
Policies and Procedures
To ensure safe and efficient operations we have a set of policies.
General Policies
- Please be punctual. If you are going to be late, contact a staff member. We will make every effort to complete the experiment, but we may need to reschedule the appointment when the workload permits.
- Plan accordingly, as the entire amount of time scheduled will be billed regardless of the amount of actual time used. If you underestimate the time needed to complete your experiment, you will be billed for the entire amount of usage. Depending on the workload, you may be asked to schedule a later time to finish your experiment. Previously scheduled appointments will take first priority.
- It is encouraged that appointments for acquisition and analysis be booked at least 24 hours in advance to ensure staff and/or instrument availability.
- Cell sorting experiments on the BD Aria need to be discussed with Core Staff prior to scheduling a time. Sorting appointments must be booked one week in advance. The Facility Supervisor must approve exceptions.
- Make sure that potentially biohazardous experiments are approved by the institutional safety board.
Training
Independent usage of the Canto II analyzer and Miltenyi AutoMacs is encouraged, and training on these instruments is provided by Core Facility Staff. Interested investigators are required to contact Core personnel to schedule this training. Individuals requesting training are required to view a web-based introduction to flow cytometry presentation prior to beginning the hands-on instrument training. Basic instrument training is generally spread out over three days, two hours each day.
Sample Delivery
- Samples to be processed by Core Facility Staff are to be hand delivered to the Flow Cytometry Core Facility located in MDC 3111.
- Cells to be analyzed must be in a single cell suspension and be free of debris and clumps, which can clog the instrument nozzle. If the nozzle becomes plugged due to poorly prepared samples, it must be cleaned and the instrument realigned. A significant delay may result, causing scheduling problems to occur.
- Samples should ideally consist of 1 x 10 6 /ml concentration, re-suspended in approximately 0.5 - 1 .0 ml of buffer, free of calcium and magnesium. This allows for the sample to be acquired in approximately 30 seconds (minimum acquisition of 10,000 events). It is understood, depending on the experiment, the number of cells is not always available. However, samples with lower concentrations will increase the amount of time required to process each sample.
- Cells must be delivered in covered tubes, which are compatible with most BD Biosciences flow cytometers. The recommended tube is a Falcon 2054 polystyrene tube with cap. The core facility will allow new users to borrow a supply of tubes until they can order their own. In the interest of time, investigators are required to transfer samples into the appropriate tubes prior to delivering them to the laboratory.
- Samples to be processed by Core Staff must be properly labeled. Without proper identification of samples, Core Staff may not be able to perform adequate acquisition and analysis.
- It is essential that any potential biohazardous materials in the samples be identified at the time the samples are delivered to the facility. The flow cytometer may create aerosols. If hazardous materials are present in the sample, safety issues must be addressed. No samples containing radioactive material are to be brought to the Flow Cytometry Core Facility.
Publications
Please, all publications based on work conducted in the Flow Cytometry Core Facility acknowledge the facility as follows:
This work has been supported in part by the USF COM’s Fred Wright Jr Flow Cytometry Core Facility.