To ensure safe and efficient operations we have a set of policies.

General Policies

1. Please be punctual. I understand that the 5 min centrifuge step is really 8 minutes, including the placing tubes in the centrifuge and discarding supernatant, hence the first time nearly everyone underestimates how long the sample prep will take. If you are going to be more than 10 minutes late, please send an email. We will make every effort to complete the experiment, but we may need to reschedule the appointment when the workload permits. When the next scheduled person shows up and we are not done, yet, we will shift to the next experiment
2. 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.
3. Schedule when you start thinking about running an experiment. This is a one person operation and I do go off on longer trips with no access to communication.
4. Make sure that potentially biohazardous experiments are approved by the institutional safety board.

Training

Independent usage is encouraged, and training on instruments is provided by Core Facility Staff. Interested investigators are required to contact Core personnel to schedule this training. If your experiment is a one off, or you run it infrequently, the core can take care of the experiment fully, but if you run it yourself, you have more ownership of it. It is very rare that users get trained to sort independently. It is much more involved that analyzers and make no sense unless you do it all the time.

Sample Delivery

1. Samples to be processed by Core Facility Staff are to be hand delivered to the Flow Cytometry Core Facility located in MDC 3111.
2. Cells to be analyzed must be in a single cell suspension and be free of debris and clumps, which can clog the instrument nozzle. This is particularly troublesome for sorts. 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.
3. Samples should ideally consist of 1 x 10 6 /ml concentration, re-suspended in approximately 0.25 - 0.4 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. If you have a lot of cells, it can be good to have them still in small volume, but to have extra media that they are suspended in at hand, so that they can be diluted if needed.
4. 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.
5. 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.
6. 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.