Approvals 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 FACScan through the on-line scheduler. Requests for use of all other cytometers must be made by contacting the Facility personnel. Please be aware that occasionally the facility is booked more than one week in advance.

Cell Suspensions

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 FACScan and the LSR II must be provided in 12x75 mm polystyrene (Falcon #2054) tubes. 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.

The recommended sample concentration for the FACScan and the LSR II

is 1x10⁶ cells per ml in 0.5 ml of 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 5x10⁶ cells

per ml in 1ml of PBS (no Mg⁺⁺, Ca⁺⁺ or serum) or fixative.

Samples derived from human sources are not accepted unless they are fixed or previously approved by the Facility Manager. All samples must be free from infectious agents. When bringing samples for multi-parameter analysis, single positive controls must be provided for each color used.

Restrictions

The FACSAria and the Reflection use 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.

Therefore, live human tissue will not be accepted for sorting or analysis unless it is certified to be free from infectious agents.

Accounts

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.

See Rates

Consultation Requirements

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.

Publishing Data

It would be appreciated if any publication which includes results of data produced from the facility that the Cytometry Core Facility is mentioned in the acknowledgement section and that a copy of the paper be provided to the facility for filing. Please see Publishing Data.