Samples must be provided in a 12 x 75 mm polystyrene tube (Fisher brand # 14-9563A or equivalent). 

The recommended cell concentration is 1 x 10 6 / sample but 5 x 10 5 /sample is the minimum required.  Cells should be suspended in a volume of 0.5 ml - 1.0 ml.

Always provide a negative control of cells only (unstained/untransfected).  For multiparameter analysis a single positive control must be provided for each color used.

Sample Collection:

Samples may be collected in any of the following (to be provided by the investigator):

12 x 5 mm *polypropelene tubes (1-4 populations at a time. Fisher Brand # 14-9561-D or equivalent)

15ml conical tubes

6-96 well plates

*Polypropylene tubes are recommended for collection from the FACSAria.  The Aria charges the stream to sort cells and a charge may build around polystyrene tubes as the cells are sorted.  Initial samples may be in either polystyrene tubes or polypropylene tubes. 

When using tubes for collection, it is a good idea to coat them with BSA, especially the 15 ml conical tubes.  This will help keep the cell droplets from sticking to the sides of the tube and drying out.  To coat the tubes, fill them with 4% BSA (bovine serum albumin) in 1X PBS (filter-sterilized) and incubate at 4 o C for at least 1 hour prior to sorting.  Just prior to bringing the tubes for sorting, pour out the BSA (this may be reused for up to 1 month), and then add the media.

How to Choose the Correct Flourochromes

Keep in mind that many fluorochromes will have overlapping emission spectra, and may be difficult to distinguish when used together.  Be sure to check the excitation and emission spectra of the fluorochromes you plan to use to optimize your experiment.  The BD Biosciences Fluorescence Spectrum Viewer may help you determine whether the fluorochromes you want to use will have a significant overlap.  We also have a list of some fluorochromes, dyes and fluorescent proteins you may check.  

An account (SAP#) or grant number (with 3-digit fund #) and slot number is required at the time of each run.  Please do not ask us to look up your account number.

To save data from the FACSAria, please bring a writeable CD, a USB portable storage drive, or a Zip disk.  Data from the FACSCalibur will be saved in a folder for you on resnet.  

The Flow Cytometry Core Facility within the Department of Microbiology and Immunology provides service for research use only.  Clinical samples or bio-hazardous material will not be accepted.  Please read the statement at the end of the submission form before signing it or submitting samples.