General Instrument Protocols and Procedures

Below are listed some of the general things we wish users to observe when using the analytical flow cytometers. Please try to observe these so that we can maintain a low down time on the instruments and to make access to all users as convenient as possible.

For instrument-specific protocols, go to: CyAn or LSR IIm, Stratedigm (under construction - see Director or Barry Udis for training)

Biohazards - Biohazardous materials are not permitted in most of the Facility locations. Use of biohazardous materials on the FACSCalibur, CyAns, MoFlo, and MoFlo XDP are strictly prohibited. Documented use of these materials on these machines, after one warning - (# of occurrences is based on a laboratory not a single user) a second occurrence will result in suspension of access to the Facility for your entire laboratory and reporting to EHS. Biohazardous materials may be used (after consultation with Facility Director) on the FACScan and on the Reflection. See Biosafety for more info. We require a copy of your Schedule F for all biohazardous material brought into the facility.

Radioactive Materials - Radioactive materials are not permitted anywhere in the Facility.

Food and Drink - No food or drinks are permitted at the machines.

Sign-up for appointments - By signing up for appointments you guarantee yourself access when you want it. It also helps us keep track of use more efficiently - we always know when you used it it just takes us longer. The new scheduling calendar allows you to add or edit your time at any time. Within 36 hours of your scheduled appointment you will not be able to delete or shorten your time or move it to another day (see Instrument Schedules for more info).

Login to Instruments: All the cytometers require users to login. Enter your ONYEN and Password. Be sure you have selected to login to the AD domain.

Log off the instrument - Please log off the machine when you are finished. You may be charged for extra time if you do not.

User Courtesies: Please try to be considerate of other users. In this regard, try to start and finish on time - the rule is you must get off when your time is up subject to negotiation with next person, always sign-up on-line to use the machine (NB: on-line Calendar is now current to the minute if people sign-up - see Scheduling Instruments), show up for your appointments or call us to let us know you can't make it.

Sample tubes - Please observe the requirements for bringing your samples in the proper tube. The FACScan and FACSCalibur require Falcon 12x75mm polystyrene plastic tubes (polystyrene is the very clear type plastic) - the CyAns require Falcon 12x75mm polypropylene (slightly cloudy) plastic tubes. On the FACSCalibur , FACScan, and CyAns you may also use the small tubes designed to fit in a 96-well format holder. For the analyzers you must bring your own tubes - we do not provide them. The MoFlo and MoFlo XDP requires 12x75mm tubes for samples but they may be either polystyrene or polypropylene (polypropylene tubes are much better for you cells)- (the MoFlo XDP can use other size tubes as well from 12x75mm to 50ml tubes - we recommend however that you use either 12x75mm or 17x100mm tubes).

Sort receptacle tubes - if you are sorting you need to provide (usually) either 12x75mm or 17x100mm polypropylene tubes. Polystyrene tubes are not acceptable as they accumulate charges during sorting which can result in large losses of sorted cells. For the 12x75 tubes place 1ml of media in the tube - when using the 70um tip we can place 3x10e6 cells in the tube - when using the 100um tip we can place about 1x10e6 cells in each tube. When using the 17x100 tube increase the numbers above by 3 fold.

Saline squirt bottles - Please do not use saline squirt bottles around the machines especially for washing off sample probes. Use distilled water. The use of saline creates a big mess in salt deposits on and around the instruments.

Clean up your mess - As the old saying goes "we are not your mother". Please clean up spilled fluids and throw away or take back to your lab materials such as aluminum foil, plastic wrap, ice buckets, pipettes, pipette tips etc. If when using the FACScan you spill a biohazardous sample you must notify us and use the provided cleanup kit to clean up the spill.

Instrument Clogs - the tubing diameters through which your cells pass are very small. Be sure your samples are free of clumps and large debris - generally if you can see you can't run it. You may have to filter your cells through a fine nylon mesh (Nitex) to remove clumps/debris. You may purchase 30um mesh filters from the Facility (we sell them strictly at cost - no markup) or prepare your own Nitex for filtering. We reserve the right to charge for removing clumps from the machine in addition to the lost machine time.

Clean the machine - Please clean the sample probe and line when you are finished with the instrument. Each instrument has a specific cleaning protocol - please see the Instrument Protocol Page for the instrument you are using for the details.

Disk space - Users are encouraged to use the data server ( \\locust.med.unc.edu\vol2\Microbiology\Groups  - Flow Facility \ youronyen) for saving data. See instructions in the Data-Saving and Transfer link. If you store anything but databases and protocols to a local drive this data could be deleted at anytime. Any user data found on the C: drive will immediately be deleted. Do not store databases, protocols, or other personal data on the data server drives - it will be deleted when space limitations require us to delete data. We will save your data for 3 months. After that, you are responsible for maintaining long-term data storage.