Cytometer Usage Rules

Training and Access:

New users should contact Ewa Menet to be able to book their times through http://www.schedulebook.com/. The following information is needed and should be provided on the NEW USER SIGN-IN SHEET (available in room S613), or by e-mailing Ewa.Menet@yale.edu:
Principal Investigator's Name User's Full name and e-mail address,
PTAEO (number for charging purposes)with expiration date,
User ID and Password of your choice (up to 15 characters, case sensitive)
The same password will be used to log in to FACStations.
To access the FACS room on evenings/weekends a key needs to be ordered (one per lab). Please contact Yale Immunobiology Business Office at 785-5211.
All new users, regardless of previous FACS experience must take appropriate training with Ewa Menet on the proper use of the instruments and Core policies.
It is important to log off after using the cytometer. If you do not, you will be charged for the time that someone else may be using.
Users who do no arrive within 30 minutes of their start time forfeit that time. Anyone may use the cytometers after this 30-minute grace period and may not be asked to leave.
A single person cannot sign up for more than 8 hours per week of "prime time" on either of the machines (i.e. no more than 8 hours combined). Prime time is Monday-Friday 12N-8PM. An unlimited amount of time may be reserved out of "prime time".

No more than 4 hours of "prime time" can be reserved in any single block. If more than 4 hours of time is needed, individuals should sign up so as to start before "prime time" or end after it, or use the weekend time.
Users will now be charged for the greater of time scheduled or actual time used on the FACS machines. Most users know that they were only being charged for the time they actually used. This new rule should discourage signing up for excessive time which previosly was a cost-free way of reserving the machines.
Individuals who do not show up for a scheduled time will be charged triple for missed scheduled time. If you are not going to be able to make a scheduled time you must delete the event from Schedulebook by 11:00 am of the day of scheduled event. On our randomly selected day 7 of the 20 unused scheduled hours were the result of "no shows".
The use of the aliases will result in a loss of FACS facility privileges for two weekes. Use of the FACS ID of others (i.e. an alias) in order to schedule extended blocks of time (greater than the allowed 4 hour blocks Mon- Fri 12:00-8:00pm)will not be tolerated. The user who scheduled time with an alias as well as the individuals who lent their ID will both have their FACS privileges suspended. It is important that you do not share your user ID and password with anyone. This rule will allow for more fair allocation of FACS time and in addition will prevent untrained users from having access to the machines. If you need to have a new password or ID issued and/or need training please contact Ewa Menet at Ewa.Menet@yale.edu or 5-7958. Also contact Ewa if your experiment requires more than the allotted four hours of prime time per day for special permission.
Maintenance and Care of Machines:
It is strongly advised to use nylon mesh to filter your single cell suspensions before staining them. Unfiltered samples will clog the machine and will interrupt your experiment. You may get a clog that will not be fixable and you may lose your experiment. For filtering, you can use the following products:
B-CMN-40 (by sq. feet) from Small Parts, Inc.:
Cell Trics sterile filter caps from Partec,
Falcon 2235 tubes with mesh in the caps from Becton Dickinson, etc.
All samples to be run on the LSRII must be filtered at the machine just before running them. This can be accomplished with BD filter tubes or with pieces of Nitex and transfer pipettes (a much cheaper solution). Anyone found running samples without a means to filter will be given a Level II violation. Repeat offenders can lose privileges to use the facility and their PI will have to pay fines.
All users must run bleach to clean and sanitize the machine (5 min 10% Clorox / 5 min Distilled Water) after finishing the experiment. You must also empty the waste tank, and put 300 ml of 100% Clorox into it and refill the sheath fluid tank. Do not under any circumstance leave the machine running or allow it to run out of sheath fluid. This causes air bubbles in the systems that are hard to fix.
It is recommended to collect your data to the hard drive of the FACS stations and then at the end of the experiment copy it to a ZIP disk or record a CD. (DO NOT RECORD DIRECTLY TO A ZIP OR CD YOU WILL LOSE DATA). Brief instructions on CD writing software are available near the instruments, and new CDs for everyone's use are located on the shelves above the cytometers.
Do not keep data files on the hard disk for more than one week - files left on the hard disk will be deleted.
Calibration of the cytometers, including Time Delay of the FACS Calibur, is being performed every morning Monday-Friday. It is strongly recommended that you do your own Time Delay Calibration if you plan to use the second laser of the FACSCaliburs.
To report any problems with the FACS, please first try to contact Geoff directly. If he is not available, uses the forms provided on the top of the cytometers and leave it on the desk with the printer.
Serious abuse of the cytometers and the above rules will be penalized. In particular, violations of rule 8 will not be tolerated, nor will bringing samples that cause clogs (see rule 7). We can and will track who leaves the machines in improper conditions.

Level I Violations are:
1: Leaving the Machine in the Run Position
2: Leaving an Overfilled Tube on the Machine (more than 1-2 ml H2O)
3: Altering the Machines Without Authorization (i.e.- removing SIP Sleeves or Loader Cylinder)
4: Any Gross Abuse of the Machines

Level I Fines are:
First Violation: $100 Fine
Second Violation: $200 Fine and FACS Facility Privileges Suspended For 2 Weeks
Third Violation: $300 Fine and FACS Facility Privileges Revoked

Level II Violations are:
1: Not Emptying the Waste Container
2: Not Filling the Sheath Fluid Container
3: Not Properly Preparing Samples - Clogging the Machine Repeatedly
4: Not Depressurizing the System After Use

Level II Fines are:
First Violation: Warning to User and PI Via E-mail
Second Violation: $100 Fine
Third Violation: $200 Fine and FACS Facility Privileges Suspended For 2 Weeks
Fourth Violation: $300 Fine and FACS Facility Privileges Revoked



		Section of Immunobiology, Department of Internal Medicine and Yale Cancer Center

Printer friendly version Cytometer Usage Rules Training and Access: New users should contact Ewa Menet to be able to book their times through http://www.schedulebook.com/. The following information is needed and should be provided on the NEW USER SIGN-IN SHEET (available in room S613), or by e-mailing Ewa.Menet@yale.edu: Principal Investigator's Name User's Full name and e-mail address, PTAEO (number for charging purposes)with expiration date, User ID and Password of your choice (up to 15 characters, case sensitive) The same password will be used to log in to FACStations. To access the FACS room on evenings/weekends a key needs to be ordered (one per lab). Please contact Yale Immunobiology Business Office at 785-5211. All new users, regardless of previous FACS experience must take appropriate training with Ewa Menet on the proper use of the instruments and Core policies. It is important to log off after using the cytometer. If you do not, you will be charged for the time that someone else may be using. Users who do no arrive within 30 minutes of their start time forfeit that time. Anyone may use the cytometers after this 30-minute grace period and may not be asked to leave. A single person cannot sign up for more than 8 hours per week of "prime time" on either of the machines (i.e. no more than 8 hours combined). Prime time is Monday-Friday 12N-8PM. An unlimited amount of time may be reserved out of "prime time". No more than 4 hours of "prime time" can be reserved in any single block. If more than 4 hours of time is needed, individuals should sign up so as to start before "prime time" or end after it, or use the weekend time. Users will now be charged for the greater of time scheduled or actual time used on the FACS machines. Most users know that they were only being charged for the time they actually used. This new rule should discourage signing up for excessive time which previosly was a cost-free way of reserving the machines. Individuals who do not show up for a scheduled time will be charged triple for missed scheduled time. If you are not going to be able to make a scheduled time you must delete the event from Schedulebook by 11:00 am of the day of scheduled event. On our randomly selected day 7 of the 20 unused scheduled hours were the result of "no shows". The use of the aliases will result in a loss of FACS facility privileges for two weekes. Use of the FACS ID of others (i.e. an alias) in order to schedule extended blocks of time (greater than the allowed 4 hour blocks Mon- Fri 12:00-8:00pm)will not be tolerated. The user who scheduled time with an alias as well as the individuals who lent their ID will both have their FACS privileges suspended. It is important that you do not share your user ID and password with anyone. This rule will allow for more fair allocation of FACS time and in addition will prevent untrained users from having access to the machines. If you need to have a new password or ID issued and/or need training please contact Ewa Menet at Ewa.Menet@yale.edu or 5-7958. Also contact Ewa if your experiment requires more than the allotted four hours of prime time per day for special permission. Maintenance and Care of Machines: It is strongly advised to use nylon mesh to filter your single cell suspensions before staining them. Unfiltered samples will clog the machine and will interrupt your experiment. You may get a clog that will not be fixable and you may lose your experiment. For filtering, you can use the following products: B-CMN-40 (by sq. feet) from Small Parts, Inc.: Cell Trics sterile filter caps from Partec, Falcon 2235 tubes with mesh in the caps from Becton Dickinson, etc. All samples to be run on the LSRII must be filtered at the machine just before running them. This can be accomplished with BD filter tubes or with pieces of Nitex and transfer pipettes (a much cheaper solution). Anyone found running samples without a means to filter will be given a Level II violation. Repeat offenders can lose privileges to use the facility and their PI will have to pay fines. All users must run bleach to clean and sanitize the machine (5 min 10% Clorox / 5 min Distilled Water) after finishing the experiment. You must also empty the waste tank, and put 300 ml of 100% Clorox into it and refill the sheath fluid tank. Do not under any circumstance leave the machine running or allow it to run out of sheath fluid. This causes air bubbles in the systems that are hard to fix. It is recommended to collect your data to the hard drive of the FACS stations and then at the end of the experiment copy it to a ZIP disk or record a CD. (DO NOT RECORD DIRECTLY TO A ZIP OR CD YOU WILL LOSE DATA). Brief instructions on CD writing software are available near the instruments, and new CDs for everyone's use are located on the shelves above the cytometers. Do not keep data files on the hard disk for more than one week - files left on the hard disk will be deleted. Calibration of the cytometers, including Time Delay of the FACS Calibur, is being performed every morning Monday-Friday. It is strongly recommended that you do your own Time Delay Calibration if you plan to use the second laser of the FACSCaliburs. To report any problems with the FACS, please first try to contact Geoff directly. If he is not available, uses the forms provided on the top of the cytometers and leave it on the desk with the printer. Serious abuse of the cytometers and the above rules will be penalized. In particular, violations of rule 8 will not be tolerated, nor will bringing samples that cause clogs (see rule 7). We can and will track who leaves the machines in improper conditions. Level I Violations are: 1: Leaving the Machine in the Run Position 2: Leaving an Overfilled Tube on the Machine (more than 1-2 ml H2O) 3: Altering the Machines Without Authorization (i.e.- removing SIP Sleeves or Loader Cylinder) 4: Any Gross Abuse of the Machines Level I Fines are: First Violation: $100 Fine Second Violation: $200 Fine and FACS Facility Privileges Suspended For 2 Weeks Third Violation: $300 Fine and FACS Facility Privileges Revoked Level II Violations are: 1: Not Emptying the Waste Container 2: Not Filling the Sheath Fluid Container 3: Not Properly Preparing Samples - Clogging the Machine Repeatedly 4: Not Depressurizing the System After Use Level II Fines are: First Violation: Warning to User and PI Via E-mail Second Violation: $100 Fine Third Violation: $200 Fine and FACS Facility Privileges Suspended For 2 Weeks Fourth Violation: $300 Fine and FACS Facility Privileges Revoked back to top