General User Guidelines
The following outlines the general user guidelines applicable to the flow core facility. All users are expected to read these and be aware of general safety standards for safe laboratory practice. Although all sample runs in the flow core facility are assisted by the operator, there are still guidelines that should be followed by all users. Please review all user guidelines before bringing any samples and be sure to follow them. If you are unsure of how to follow these guidelines or have any questions at all, please contact the operator. Failure to follow these guidelines can result in damage to the machine and therefore will result in loss of facility privileges.
1. Please complete either a new user request for BDAccuri C6 users or a sort request for BDFACSAria II users before attempting to reserve any time on either instrument. If the operator does not have information for you on file, your reservation request will be rejected.
2. Please make sure all information on your form is up-to-date and accurate. We, too, are bound by health and safety regulations and are responsible for ensuring that only the proper sample types are being admitted to the facility. For this, we rely on your honest, accurate reporting on your request forms. Please give us the correct information so we know what is in the samples we are handling!
3. If you have filled out the correct form but some minor information has changed, such as fluorochromes to be used, sample concentration, nozzle size, etc., then please simply notify the operator. (Please note that these factors greatly affect the machine configuration and set-up. If you do not notify the operator of any changes, your run will be set-up according to the information received on your form. If you arrive and indicate that you would like different parameters, i.e. the 85um nozzle rather than the 70um nozzle, the machine will have to be totally re-configured. This process can take 30 minutes or more. You will be billed for this time, and if there is someone scheduled after you, your time cannot be extended).
4. If you have filled out the correct form but some major information has changed, such as cell type, infectious agents, transformation, etc., then please fill out a new form. These are factors we need to be aware of
5. Please be sure to check that the fluorochrome(s) you intend to use are compatible with the machine you would like to reserve. If you are unsure, please ask the operator. Failure to confirm fluorochrome-instrument comparability could mean that your sample will not show any fluorescence on the instrument you choose. Unfortunately, you will still be billed for this unsuccessful run. You can avoid this by checking before you bring in any samples. Click here to view the BD SpectraViewer for additional assistance.
6. There is limited space available in the facility for minor sample preparation, including dilution and centrifugation. There is not space for sample dissaggregation, tissue preparation, etc., nor is this appropriate. Samples should be ready upon arrival at the facility.
7. Please bring your own collection tubes/plates of the appropriate size for your experiment if you are sorting. Please also bring the appropriate collection buffer.
8. Safe transport of samples to the facility and within Syracuse University academic building is of paramount importance. All samples must be transported to the facility in sturdy, leak-proof double-walled containers (at least one layers must be leak-proof) capable of preventing escape of the sample in the event of breakage or spill of the sample tube(s). The container must have a lid that fastens securely during transport to further prevent escape of sample in the event of an internal spill. Prior to leaving his/her own facility, the user should wipe down the outer surfaces of the transport container with disinfectant (for example, 10% bleach solution) to sterilize it. The outer surface is now safe to touch, regardless of biosafety level, and the user should not use gloves to carry the container to the facility. Finally, prior to leaving his/her own facility, the user should affix the appropriate biohazard warning label to the outside of the container. The label should clearly indicate the biosafety level appropriate for the samples. The container should remain securely closed until the user reaches the facility.
9. If your samples are not finished within your allotted time and there is another user after you, your run will be ended at the time necessary to prepare for the next run.
10. Please plan to either stay to wait for your samples or be back on time to collect them. We do not have the space to store your samples for you.
11. Please arrive at least a few minutes in advance of your reserved time slot. Late arrival on your part means that your run will start late, which potentially takes away from the total amount of time for your run if there are other users scheduled after you.
12. Flow cytometry (and, by extension, cell sorting) works because of difference in light scattered by lasers. If the cells in your sample are stuck together or are too concentrated, your results will be affected. Please refer to the sample preparation guidelines for more information.
13. Please make sure you know the system you are working with! Information you can provide about cell size, expected fluorescence, and the relationship between various types of cells in mixed samples can greatly aid the operator in providing you with accurate results. New users are encouraged to contact the operator regarding their experimental design before reserving any time with the instruments. The university makes no warranties, expressed or implied, as to any matter whatsoever, including without limitation, the accuracy of the results.
Please note that all sample preparation should be completed prior to arrival at the facility due both to lack of space for sample preparation and lack of necessary containment for biological samples. There are pipettes, centrifuges, and a vortex accessible to users of the facility; however, these should be used for the minimum necessary amount of sample preparation. Sample should never be uncapped on the bench top, and samples will never be vortexed in open tubes (lid must always be on and secure to prevent spills and aersolization). By preparing sample prior to arrival, users not only contribute to facility maintenance and safety standards, but also ensure that their allotted appointment begins on time.
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