The gentlemacs dissociator provides optimized programs to attain single cell suspensions from various tissues for example mouse spleen.
Single cell suspension from spleen.
Continue at step 6 15 below.
Harvest mouse spleen and prepare a single cell suspension.
Resuspend the pellet in 5 ml spleen of lysis buffer.
Preparing a true single cell suspension of the primary tissue sample will optimize cell separation from tissues by avoiding additional cell loss and enabling maximum labeling of the target cells.
Discard the supernatant and resuspend the cells in pbs w 0 1 bsa to 1 x 10 7 cells ml.
I used flow cytometry to detect single cell suspension from mouse spleen and there were too many dead cells.
Aim of this experiment is to isolate single cell suspension from the spleen and thymus of the mouse.
Red blood cell lysis after having prepared the single cell suspension.
Spleen and thymus are the lymphoid organs of an animal starting from lower vertebrate like fish to higher vertebrate like human.
6 12 transfer several tissue pieces to strainer and keep moistened with d pbs.
6 11 set up another cell culture dish with 5 10 ml d pbs.
This video outlines how to harvest cells from a primary tissue sample and prepare a single cell suspension prior to performing cell isolation.
Pellet the cells by centrifugation 300 400xg at 2 8 c and aspirate the supernatant.
6 10 filter the cell suspension through a sterile cell strainer to remove dispersed cells from tissue fragments.
This video demonstrates how to harvest cells from a spleen sample and prepare a single cell suspension prior to performing cell isolation.
Lysis of mouse erythrocytes red cell lysis.
In the case of peripheral blood and lymph the cells are already in single cell suspension and the techniques for the separation of the various populations and subpopulations of cells are also discussed.
This protocol describes how to harvest cells from a spleen sample and how to prepare a single cell suspension prior to performing cell isolation.
Preparing a single cell suspension of the primary tissue sample will optimize cell separation by avoiding additional cell loss and enabling maximum labeling of the target cells.
Peripheral blood lymph and lymphoid organs.
6 13 open a sterile 3 or 5 ml syringe.
Therefore culture of cells derived from these lymphoid organs will help to understand many aspects in the field of immunology.
