Flow cytometry (FACS)

/ print page

<<< Service >>Target identification and validation >> Flow cytometry (FACS) >> Immunhistochemistry >> Western blot >> RealTime-PCR >> Microarray analysis >>

Flow cytometry is a powerful analytical tool that enables the characterisation of cells on the basis of size and granularity (light scatter characteristics) and a number of different parameters defined by fluorescent antibodies and dyes. Cell populations from in vivo tissues or in vitro cultures can be analysed. Cells must be suspended either by enzymatic or mechanical dissociation. Commonly investigated cell surface antigens in our laboratory are markers of the haematopoietic lineage either for characterisation of cultured cells or for determining the differentiation status of engrafted human cells in immunodeficient mice.

Human-specific monoclonal antibodies are for example:

Stem cells:
T-cells:
B-cells:
Granulocytes:
Monocytes:
Erythrocytes:
Integrins:
Selectins:
Immunglobulin superfamily:

CD34, CD38, CD133 and CXCR4
CD3,CD4 and CD8
CD19 and CD20
CD15
CD14
CD235a
CD11a, CD18 and CD49d
CD62L and CD62P
CD56, CD54 and CD31

Analysis of relative cellular DNA content of cells within an actively-cycling cell population using the fluorescent DNA dye propidium iodide. CD34+ stem cells are stimulated with stem cell factor in vitro and the cell cycle analysis was performed in cells cultured for seven days.

Engrafted human cells in mouse bone marrow. Human CD34+ stem cells were i.v. injected into NOD/SCID mice. Eight weeks after transplantation mice were sacrificed and the isolated bone marrow cells were stained with HLA-I. 30% of mouse bone marrow cells were HLA-I positive.

Cytokine-producing splenocytes from mice: NK-cells from spleens were enriched and either non-stimulated or stimulated by PMA, fixed, permeabilised and stained with APC-CD49b and PE-anti-mouse IFN-γ according to BD Biosciences intracellular cytokine staining protocols.

Top