Single Cell Projects

Our genomics core has performed hundreds of single cell captures since implementing 10x Genomics techniques in our core and we are happy to work with you to figure out the best way to set up your project. To ensure we have the necessary reagents available and staff members on-site to cover out-of-hours captures, we ask that our clients follow our scheduling procedure below so we can ensure consistent coverage of everyone's projects. If you have a project that poses particular difficulties (eg. short notice, late hours, BSL3), please contact us well in advance to make arrangements.

Scheduling

To schedule your single cell capture, please follow the steps below:
  1. Email Kathryn Pellegrini at least one month in advance with your requested date/time and number of captures. If you have unavoidably short timelines, please call us to discuss your project.
  2. Once your capture has been confirmed you will receive a calendar invite for the specified date and time. If you have not received a calendar invite, the capture has not been scheduled.
  3. Fill out our online project submission form at least 24 hours in advance of your capture. Samples will not be accepted without a completed project form.
  4. Keep us updated on the day of the capture by email in case there are any changes to the timeline (please email all staff members included on the calendar invite).

 

Sample Preparation Information

For best results, single cell RNAseq captures should be performed on freshly collected cells so it is important to schedule these experiments in advance. If you have not previously done any single cell experiments with us, please contact Kathryn Pellegrini to discuss your project.
  • Cells should be provided as single cell suspensions in a compatible media. See the 10x Genomics Cell Prep Guide for more details. We frequently receive cells in RPMI + 10% FBS for 10x captures.
  • If your cell preparation contains red blood cells, consider using a RBC lysis protocol. The 10x device will capture all types of cell that are loaded.
  • Please provide us with either a cell count from a hemocytometer or the number of cells that you have sorted to give us an idea of what is in the sample.
  • Once we receive the samples, we will perform a count and check the viability if possible (for samples with at least 20000 cells). If the viability and count indicate a high likelihood of an unsuccessful capture, we will contact you to discuss options.  Please make sure that you provide us with a contact number for you or your PI so that we can discuss any issues immediately.
  • Handle cells gently (no rough pipetting or harsh vortexing) and keep them on ice at all times they are not being actively worked with.