Single Cell Suspension From Spleen

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.

Single cell suspension from spleen. Aim of this experiment is to isolate single cell suspension from the spleen and thymus of the mouse. This chapter discusses that lymphoid cells may be obtained from various sources. 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. Peripheral blood lymph and lymphoid organs.

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. Spleen and thymus are the lymphoid organs of an animal starting from lower vertebrate like fish to higher vertebrate like human. Lysis of mouse erythrocytes red cell lysis.

6 13 open a sterile 3 or 5 ml syringe. Red blood cell lysis after having prepared the single cell suspension. Pellet the cells by centrifugation 300 400xg at 2 8 c and aspirate the supernatant. Harvest mouse spleen and prepare a single cell suspension.

Discard the supernatant and resuspend the cells in pbs w 0 1 bsa to 1 x 10 7 cells ml. Continue at step 6 15 below. Resuspend the pellet in 5 ml spleen of lysis buffer. Single cell suspensions are a prerequisite for many experiments for example to achieve the highest possible purity and recovery during cell separations with macs technology.

This video demonstrates how to harvest cells from a spleen sample and prepare a single cell suspension prior to performing cell isolation. 6 10 filter the cell suspension through a sterile cell strainer to remove dispersed cells from tissue fragments. This video outlines how to harvest cells from a primary tissue sample and prepare a single cell suspension prior to performing cell isolation. 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.

The gentlemacs dissociator provides optimized programs to attain single cell suspensions from various tissues for example mouse spleen. Fluorescent activated cell sorting is a common option to obtain single cells from clinical samples for single cell applications but requires 100 000 viable cells in suspension and the need for.