Approval for Flow Analysis or Cell Sorting
Approval must be obtained from the Facility Manager for cell analysis or sorting. After an initial consultation, the user may gain access to the FACScanto II through the on-line scheduler. Requests for use of all other cytometers must be made by contacting the Facility Manager. Please be aware that occasionally the sorter is booked more than one month in advance.
Cell suspensions are typically prepared in the investigator's laboratory and brought to the Cytometry Core Facility ready to be run. There is a wet lab available in the facility with limited equipment for use in the preparation of samples, if needed.
Samples & Containers
Samples for the FACScanto II and the LSR II must be provided in 12x75 mm polystyrene BD brand Falcon tubes. The only tubes compatible with these analyzers are:
- Cat. # 35-2052 - sterile, no cap, 125/bag
- Cat. # 35-2054 - sterile, with cap, 125/bag
- Cat. # 35-2058 - sterile, with cap, 25/bag
- Cat. # 35-2008 - nonsterile, no cap, 1000 bulk
- Cat. # 35-2235 - sterile, with cell strainer, 25/bag
Samples for the FACSAria may be provided in 12x75mm polystyrene tubes or 15 ml conical tubes.
It is the responsibility of the investigator to be certain that the cells are aggregate free. It is recommended that the samples be filtered using 30um mesh to remove clumps prior to bringing samples to the lab. Unclogging the instruments can be very time consuming and the users will be charged for that time.
The recommended sample concentration for the FACScanto II and the LSR II is 1x106 cells per ml in PBS (no Mg++, Ca++ or serum) or fixative.
1-3% paraformaldehyde is the recommended fixative for most cell types.
The recommended sample concentration for the FACSAria is 3x106-5x106 cells per ml in PBS (no Mg++, Ca++ or serum) or fixative.
Pathogenic samples or live samples derived from human sources must be approved by the Facility Manager. When bringing samples for multi-parameter analysis, single positive controls must be provided for each color used.
Turn-around time is dependent on the length of the appointment. In general, samples are processed and data is received by the user at the end of their appointment time.
The FACSAria has a jet-in-air fluidics system, which potentially produces aerosols, especially when sorting. These aerosols may contain infectious agents, such as hepatitis or HIV, when sourced from live human tissue. In addition, sorting known infectious agents such as bacteria or cells transfected or transformed with infectious agents also pose a risk from aerosol exposure. Since this aerosol is under pressure, potential for infectivity is theoretically much higher than incidental exposure during analysis.
The addition of a Baker BioProtect III biosafety cabinet, in which the FACSAria is now housed, allows for sorting of certain live human tissue and BSL2 agents. Approval for BSL2 sorting must be obtained through the Core Facility Manager.
An account or grant number must be included with each sample run. Charges are based on 15 minute increments for analysis or sorting. It is the responsibility of each user to keep his/her User Profile updated regarding account information, phone number, and email address.
Because of the complexity of the FACSAria, there is a one hour setup charge for each sorting session.
If you are not affiliated with the Arizona Universities, please contact the Cytometry Core Facility for an initial consultation and service quotes.
We provide personalized services. A consultation is required before the first scheduled session to discuss controls needed and to expedite any special requests. This can easily be accomplished by contacting the Facility Manager.
It would be appreciated if the Cytometry Core Facility is acknowledged when publications include data obtained from the Facility and that a copy of the paper be provided to us for filing. Please see Publishing Data.