Our high speed cell sorter (FACSAria) can perform 2-way and 4-way cell sorting.
Our FACSAria has 3 lasers: blue-488nm, red-633nm, and violet-407nm
A yellow laser of 561nm will be installed soon.
Please refer to the following table to select your Fluorophores. If you have to use any other fluorophores outside the following spectra, additional filter has to be purchased.
FACSAria Filter configuration
Dichroic Mirror (nm)
|Bandpass Filter (nm)||Intended Dye|
|Red (633nm)||735||780/60||APC-Cy7, APC-H7|
|Violet (405nm)||502||530/30||Alexa Fluor 430|
Alexa Fluor 405
Texas Red, PI
Cells for Sorting
• Cells for sorting must be filtered using Falcon 12 x 75 mm polystyrene test tubes with cell strainer
Snap cap (Falcon #2235). Filter your samples inside the culture hood right before bring to sort.
• Minimum sample volume is 0.5 ml.
• Cells should be concentrated to at 10- 20 million cells/ml for the high speed sorters. Ex. Lymphocytes
– concentrate can be higher (up to 50 million cells/ml) Ex. Sticky/Adherent cells – concentrate at
maximum 20 million cells/ml
• Place all the cell tubes and collecting tubes on ice
1) A negative control: unstained sample or a sample stained only with the secondary Ab.
2) Single color controls. If you have samples stained with more than one fluorochrome per test tube,
bring in a control sample stained with each fluorochrome individually. This is for compensation
Ex. of controls for a simple GFP experiment:
• Negative control = cells not expressing GFP
Ex. of controls for a dual FITC-PE Experiment
• Unlabeled control
• FITC only control
• PE only control
1) Use appropriate media to collect cells: put about 1 ml of collecting media in the collection tubes.
2) Use appropriate size of collection tubes.
Ex. If you expect to collect 3 million cells, use 5 ml or 15 ml tube
Ex. If you expect 100,000 cells, use a 1.5 ml eppendorf tube
3) Add antibiotics in the collecting media if sorted cells are to be cultured following cell sorting. Wash
the collected cells once after sorting. The FACSAria cell sorting system could not be 100% sterilized
unless this equipment is put inside a sterile hood.
The current setup for the cell sorter (FACSAria), which has no containment system for aerosols, prevents us from running samples that contain radioactivity or potentially infectious agents to humans.