Flow Cytometry Core Facility - New Investigators
Welcome New Investigators! Here are some quick tips and guidelines to make your first experiment with us as successful as possible!
Samples for Analysis
- Please label your tubes 1, 2, 3, etc. Make sure that the numbers are large enough so that we can read them easily. Have your tubes in numerical order.
- Please do not bring your samples in styrofoam racks.
- Please call ahead to schedule an appointment if your samples need to be run live or are time sensitive (i.e. not fixed or in ethanol waiting for PI cell cycle staining).
- Make sure that your rack is labeled well so that you can pick it up after your experiment has been run.
- The Caliburs and the LSR II require using the polystyrene BD Falcon 352008 tubes (available through the lab supermarket).
- Please have your cells at 106 cells per ml with a minimum volume of 300 µL. Un-fixed untested or known infectious samples MUST have secondary containment such as a sealable plastic or styrofoam box for transport.
- Please indicate if you need printouts of your Calibur sample analysis.
- Fill out all of the fields on our provided data sheets. (Sheets can be downloaded on our Protocols page.)
- Feel free to check with us if you have any questions regarding which fluorochromes are appropriate for our machines.
Samples for sorting
When scheduling a sort, the following information is needed:
- The biosafety level of your sample - as determined by the IBC (critical information concerning sort samples can be found on our Biosafety page).
- Are the cells potentially infectious or from human sources? Untested or known infectious samples MUST have secondary containment (capped tubes in a rack in a sealed box)!
- Total number of cells and samples to be sorted.
- Type of cells/sources being sorted (larger cells need to be run slower for optimal purity.)
- Nozzle size (70, 85, and 100µm available - cell size should not exceed 30% of the nozzle size).
- Number of cell populations you want back ( i.e. a 1, 2, 3, or 4 – way), bulk sort or into plates.
- What fluorochromes will you be using?
- Do your cells need to be kept sterile?
- What temperature your cells must be maintained at?
- Based on this information we will choose the best sorter to fill your needs.
Sorter availability varies; please call 2 weeks ahead of your desired sort date, although sorting time is often available the next day.
What to bring for a sort:
- A completed Sort Sample Request Form including IBC docket number and the IBC's designation for the biosafety level of your sample must be provided to the Flow Core at least 2 days before the first sort of a BSL-2 sample or for EACH and EVERY BSL-2+ or BSL-3 sort.
- Samples at a final concentration of 1-2 x 107 cells per ml, with a minimum volume of 1 mL.
- Additional media with 1% serum to dilute the sort sample if necessary.
- Media with 20% serum for cell collection, or other appropriate collection reagent.
- When sorting into plates, pre-fill each well with 1/2 max volume of an appropriate growth medium.
- Control samples (1 x 106 cells each): a negative control and a positive single control for each fluorochrome being used.
- All of the above in an ice bucket, unless your cells prefer room temperature. Untested or known infectious samples MUST have secondary containment such as a sealable plastic or styrofoam box for transport.
- A data sheet with: name, PI name, speedtype number, phone number, and a description of the sort experiment with controls and colors listed.
Remember that if a cancellation needs to be made, we require 24 business hours notice for analysis and 48 business hours notice for sorting to avoid being charged for the time.
For machine specific information, visit our Instrumentation page.