Sample Preparation Guidelines

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.

BDAccuri C6 Sample Preparation Guidelines
1. Cells should be filtered prior to processing.  This reduces cell aggregates, which not only avoids clogs in the instrument tubing but also ensures the appearance of singles cells and the collection of data only pertaining thereto.
2. If cells are typically aggregated (i.e., from tissue samples), then samples should be disaggregated prior to arrival at the facility.  Trypsin is typically used, although it should be inactivated.
3. Optimal cell concentration in the Accuri C6 is 10^6 cells/mL.  However, the instrument can handle concentrations of approximately 5000-5×10^6 cells/mL, so some variation is acceptable in sample concentration.  Please note, however, that the concentration of cells in your sample will affect how quickly you will reach your target event number and thus the overall amount of time that you use the instrument.

BDFACSAria II Sample Preparation Guidelines
1. Cells should be filtered prior to processing.  This reduces cell aggregates, which not only avoids clogs in the instrument tubing but also ensures the appearance of singles cells and the collection of data only pertaining thereto.
2. If cells are typically aggregated (i.e., from tissue samples), then samples should be disaggregated prior to arrival at the facility.  Trypsin is typically used, although it should be inactivated.

Sample Transportation Guidelines
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.

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