High Speed Cell Sorter
We are pleased to announce that the CMRC High-Speed Cell Sorting Facility is open and operational. We hope that the availability of the high-speed cell sorting facility will further enhance your ability to perform excellent research.
The Facility has a BD FACSAria high-speed cell sorter equipped with 488nm, 407nm and 635nm lasers, which can measure up to 9 cell markers and sterilely separate cells into 4 tubes at one time. We have been able to routinely separate and collect 4 subsets of cells at the same time, based on sorting criteria using scattering profiles and 5 fluorescent colors. Several common fluorochromes (FITC, PE, PE-Cy5, APC, PE-Cy7) and fluorescent proteins (GFP, DsRed, HcRed) have been used successfully.
The purity of the sorted cells has been shown to be quite satisfactory and the sorted cells have been demonstrated to be functionally intact. The sorting efficiency and the maximum cell yield obtainable, however, have not been tested vigorously.
Procedure for Using the BD FACSAria High Speed Cell Sorter:
1. If you have cells that you want to sort, please contact Lei Wang (x5-6501, , C.408G) to schedule your experiments. Please complete the Experiment Information Form and send the form to Lei in advance of your experiments. A technical sheet on how to prepare your samples is also available.
2. The rate for use of the cell sorter is $50 per hour, effective May 1, 2005. A billing form should be completed before you schedule your first experiment.
3. Please understand that for a cell SORTING experiment to be successful, you will need to know beforehand the scattering profile and fluorescent pattern of your target cells. You might need to first perform flow cytometric ANALYSIS of samples of your cells and provide the sorting facility operator with the results before the sort can be set up.
4. Please expect that the sorting experiment might have to be repeated several times before the optimal condition for a successful sort could be worked out.
5. The sorted cells will be analyzed after the sort to provide you with information about their phenotype and purity, but you might need to perform your own validation assays to show that the sorted cells are really the cells that you want.
6. All sorts will be performed in a sterile condition so that the sorted cells can be cultured afterward. While we attempt to maintain the sterile condition as much as possible, contamination of sorted cells might happen and you should be aware of this possibility.
7. Because the sorting process produces aerosols that can constitute a health hazard to facility personnel, the sample restrictions listed below will be followed:
Human materials to be sorted must have documented proof of HIV, hepatitis B and hepatitis C negativity.
No infected samples or samples containing infectious materials will be sorted.
No samples containing radioactivity will be sorted.
No bacteria and yeast will be sorted.
8. Cell sorting experiments, in some cases, could be difficult and challenging. If you have scientific or technical questions about cell sorting, you can contact Dr. William Tse (5-6501, ), who might be able to provide you with some technical and scientific advice, but it will be provided only as a courtesy.