Preparing a single cell suspension is a key objective toward the goal of a perfect sample preparation. One of the first and easiest steps to do is to remove any large aggregates via filtration through 30-70 uM nylon mesh. In order to minimize sample loss follow the steps below.
- Syringe capable of holding your sample with an additional ml of capacity, along with the syringe cap.
- Aleur lock needle, typically large gauge such as an 18.
- ½” by ½” piece of nylon mesh in the 30-70 uM pore size.
- Safety razor blade
- Aspirate your sample into the syringe with the needle while maintaining 1 ml of air void volume.
- Once sample is in syringe discard needle and invert the syringe into the upright position such that the 1 ml air bubble is at the tip of the syringe.
- Maintaining the syringe in the upright position, sandwich the mesh between the syringe and the saved syringe tip, holding the mesh in place.
- Using the blade, cut off a few mm of the syringe tip creating a hole for the filtered sample to pass thru.
- Invert the syringe such that the void bubble rises to the plunger side and then push the sample gently thru the mesh along with the last ml of air, ensuring that none of the sample remains in the syringe.