# How to Prepare a Single-Cell Suspension from Primary Tissue Samples (e.g. Mouse Spleen)

**Channel:** STEMCELL Technologies
**Source:** https://www.youtube.com/watch?v=N0jftyYqM38
**Transcript page:** https://www.withtranscript.ai/video/N0jftyYqM38

## Chapters

- 0:05 — Why Single Cell Suspensions Matter
- 0:37 — Dissociating Tissue and Filtering
- 1:30 — Final Wash and Downstream Processing

## Transcript

**[0:05]:** one of the most important factors in successful cell isolations happens before you even start your separation preparing a true single cell suspension of your starting sample will optimize your cell separations by avoiding additional cell loss and enabling maximum labeling of the target cells this video outlines a method to harvest cells from a primary tissue sample and prepare a single cell suspension prior to performing cell separation when working with primary tissue samples cells must be dissociated from

**[0:37]:** the tissue in order to prepare a single cell suspension in a sterile dish containing dissociation medium mince the harvested tissue place a mesh strainer on a sterile 50 mil conical tube a 70 or 100 Micron cell strainer works well transfer the dissociated tissue and the dissociated media to the mesh strainer gently pass the dissociated tissue through the strainer wash the tissue in cells in the strainer by gently adding a few milliliters of PBS repeat this step and then discard the leftover tissue and strainer top up the tube containing the cell suspension with PBS and collect the cells by centrifugation

**[1:30]:** carefully remove and discard the supernant without disturbing the pallet gently tap the tube to resuspend the pellet You may wish to do an optional wash at this stage you should now have a single cell suspension that is ready for cell counting and subsequent processing using easysep to isolate your desired cells for more information on preparing single cell suspensions from primary tissue samples and other procedures please visit our website at stem cell.com