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• Access and Scheduling
• General User and Sign Up Rules
• Sample Preparation
• FACSCAN Data Handling And Analysis

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Access and Scheduling:

• Hours of Facility Operation:
  
• Daytime = 8:30 - 17:30 hr
• Evening = 17:30 - 23:00 hr
  

• Access Limits to LSR II, and FACScanto by Trained Personnel from User Laboratories
  
  
• Access to the booking times depends on whether or not your laboratory has defined itself as a frequent user, and has paid the frequent user access fee. (See Classification of Research Users.) The following rules apply.
  
  
• Research User Labs
• may book in advance, for any available day or evening times
• Limits are:
  • no limit on the evening schedule
  • There is a limit of 5 experiments, or 10 hours of use per week total time during the daytime schedule for each user from a given lab. For each lab, there's a limit of 6 hrs. during the daytime schedule, with two or more users from one research lab.
  • a total of no more than 8 experiments in advance, on the daytime schedule
  • no experiments will be scheduled more than 4 weeks in advance on the day schedule

• Access and Scheduling for LSR II or FACScanto with Assistance of the Facility Operator
  
  
• To book and access for samples to be run by the Operator, you must first contact the Operator. You must describe the kind of experiment, cell type, fluorochromes, etc. The Operator must be given your name and phone number, as well as, a research billing account number and department of affiliation.
• If you have never used the LSRII or FACScanto before, and are unsure about the experimental design for use of these instruments, contact the Facility Director.
• The Facility Operator is only available to run samples during the daytime hours.
• (8:00-17:00 hrs).
  

• Access and Scheduling of VantageSE Instrument
  
  
• The Vantage instrument can only be operated by a designated facility operator.
• To sign up, you must first contact the Operator and discuss with her the kind of experiment, cell type, flourescent, etc. Only after it is clear what is required will a scheduled time by assigned. The Vantage Operator will enter your name and phone number in the schedule for the agreed upon day and time. You must give a billing account number to the Operator prior to the day you use the Vantage.
• If you are uncertain if the Vantage instrument is suitable for your experiments, then first contact the Facility Director.
• The Vantage instrument is available only in daytime hours (8:00 - 17:00 hrs.)
  

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General User And Sign Up Rules:

• Users must wear a lab coat with a security badge at all times. Entry after 1700 hours may not be allowed otherwise. All evening users from any laboratory must inform the Operator of their name and security badge #, prior to the day they will use the Facility.
• Any cancellation for use of any instrument must be made at least 3 hours prior to the scheduled time. If you must cancel a LSR II or FACScanto experiment, contact the booking administrator(M-F) at or phone (x22970), and tell the Facility operator. Should this happen on a week-end call x21589 & notify person(email) booked before of after you. If cancelling a Vantage appointment, contact the Operator. This allows others to make use of the time. Failure to make a proper cancellation could result in a 1 hour penalty charge.

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LSRII, and FACScanto Booking Online and Log of User Hours:

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Sample Preparation: (For All Instruments)

• ALL samples MUST be in one of two standard running buffers:

• Buffer for Viable Cell Preparations
  
• PBS pH 7.0-7.5 - plus 1-10 mg/ml BSA (optional), plus azide (optional)
• no pH color indicator dye should be present
• exceptions MUST be discussed with the Operator or Facility Director in advance
  
  
• Fixed Cell Buffer
  
• Samples may be run with fixative, but this must be low concentration (<1% paraformaldehyde) - No Gluteraldehyde
• All samples should be washed free of unbound fluorescent labels. If the experiment requires free label, preliminary work must be done to ensure that the label will not "contaminate" the instrument (check with Facility Operator or Director)
• Samples with large visible aggregates or suspended particles MUST be filtered through nylon mesh (60 micron) prior to run. Researchers should purchase their own nylon mesh in advance and have available at the time of sample running.
• All samples MUST be in FALCON #2058 (12X75 mm) tubes, (suggested retailer). NO SUBSTITUTES ACCEPTED. These fit properly on both the FACScanto, LSRII and Vantage instruments.
• All samples brought to Facility by researchers who are running their own samples must be capped while in transport.
• All samples given to the Operator to run MUST be capped
• All sample preparation (staining, washing, fixing) must be completed before entering the Facility.

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Data Handling:

• Each laboratory will have an assigned sub-directory on each instrument computer. This is for compiling data from the current experiment. DO NOT leave data on the computer for long periods of time. Occasional "purging" of the computer hard-drive is done, with no view to saving your data. GET THE DATA OFF QUICK OR LOSE IT!
• Data can be copied to floppy diskettes or to removable ZIP disks, or writable CD. Each laboratory will have to purchase their own diskettes, ZIP or CDs for storage.
• Diskettes, ZIP or CDs must be provided by the researcher for the Facility Operator to use if data storage is requested, when the Operator runs the samples.

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