Using Sorters

USING THE SORTERS - To access sorting in our facilty, you can call 919-966-1530 to schedule sorting time on one of our instruments. We request that you complete and send a Sort Request Form by fax or email prior to your appointment.

We require a copy of your SCHEDULE F for any biohazardous materials brought into the core.

MoFlo, MoFlo XDP, Reflection

The sorters are only operated by Facility staff - the instruments are too sensitive and require extensive training for proper use. Most users do not sort often enough to become proficient or would not want to invest the time to do so. For all sorts we require that you allow us to re-analyze each sorted population to verify the sort success- we will use as few cells as possible. This is not only good science but assures us and you that we/the machine have performed satisfactorily. If this is not the case, we need to troubleshoot why you are not getting the expected results. If you have fewer than 10,000 sorted cells (in a sorted collection tube) you may request that we do not use some of them to re-analyze. If you do this we will not be responsible for the sort or subsequent experiments that you may do with the sorted cells nor will we make any billing adjustments.

Sort sterility - We rarely have sterility issues reported to us and in most cases it can be traced to a contaminated sample although we will never dismiss the possibility of a machine contamination. Please discuss with us any issues you may be having with sterility and we will be glad to help you troubleshoot the problem. We require that you include gentamicin (50ug/ml) antibiotic (in addition to your normal antibiotics) when you re-culture any sorted cells. While we have high confidence that the sorter will deliver sterile cells, we do work in a less than optimal environment that may contribute microbial contamination (primarily airborne). While this appears rare in our experience it is a possibility. We have done many sorts where subsequent culture has been performed with no antibiotics without reported contamination. Omitting the gentamicin is at your risk.

Cloning/plate sorting experiments - When we perform sorting where we place a very small number of cells (often one) into plate wells we suggest - if possible - that you prepare enough cells so we may sort a sufficiently large number 10,000-20,000 in a tube to provide a re-analysis sample. Plates must be pre-loaded with the media of your choice. For 96-well plates we suggest 100uL. We have the ability to sort many different plate configurations with 96-well and 384-well being the most requested.