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How to Prepare a Single-Cell Suspension from Mouse Spleen

[♪ music plays throughout ♪] [STEMCELL Technologies] 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 [Vicki Stronge, PhD] [Senior Product Manager] 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 [Technical Guide] [Preparing a Single Cell Suspension from Primary Tissue Samples] and prepare a single-cell suspension prior to performing cell separation. When working with primary tissue samples, cells must be dissociated from 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 mL conical tube. A 70 or 100 micron cell strainer works well. Transfer the dissociated tissue and the dissociation media to the mesh strainer. Gently pass the dissociated tissue through the strainer. Wash the tissue and cells in the strainer by gently adding a few milliliters of PBS. Repeat this step and then discard the leftover tissue and strainer. Top off the tube containing the cell suspension with PBS and collect the cells by centrifugation. [Please contact Technical Support for appropriate centrifugation speeds] for your sample, [email protected]] Carefully remove and discard the supernatant without disturbing the pellet. 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 [] [STEMCELL Technologies] [Scientists Helping Scientists] []

Reynold King

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