Flow-cytometry techniques in radiation biology
Considerable evidence exists that all blood cells are derived from HSC. These cells are of interest to radiobiologists because they are highly sensitive to low doses of ionizing radiation. Hematopoietic stem cells (HSC) are present in the marrow at a concentration of approximately 2-3 HSC per 1000 nucleated marrow cells. In the past, only clonogenic assays requiring 8-13 days and ten irradiated recipient rodents were available for assaying HSC. Because of the importance of HSC in the post-irradiation syndrome, the authors developed a new rapid method based on flow cytometry not only to assay but also to purify and characterize HSC. This new method makes extensive use of non-clonal antibodies conjugated to fluorescent phycobiliproteins through the sulfhydryls of the hinge region of the IgG molecule. An optical bench arrangement with a dye laser and an argon laser was used for dual excitation of the phycobiliprotein-monoclonal antibody conjugates and various cellular and DNA probes. Using 4', 6-diamidino 2-phenylindole dihydrochloride (DAP) exclusion to identify viable cells, it was possible to follow regeneration of post-irradiated rat marrow HSC.
- Research Organization:
- Armed Forces Radiobiology Research Inst., Bethesda, MD (USA)
- OSTI ID:
- 5927930
- Report Number(s):
- AD-A-205807/1/XAB; AFRRI-SR-88-42
- Resource Relation:
- Other Information: Pub. in Toxicology Letters, Vol. 43, 219-233(1988)
- Country of Publication:
- United States
- Language:
- English
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BLOOD CELLS
BIOLOGICAL RADIATION EFFECTS
HEMATOPOIETIC SYSTEM
STEM CELLS
BONE MARROW
DNA
DOSE RATES
DYE LASERS
IMMUNOASSAY
IONIZING RADIATIONS
MONOCLONAL ANTIBODIES
OPTICAL EQUIPMENT
PHYCOBILIPROTEINS
PROBES
RADIOBIOLOGY
RODENTS
VIABILITY
ANIMAL CELLS
ANIMAL TISSUES
ANIMALS
ANTIBODIES
BIOASSAY
BIOLOGICAL EFFECTS
BIOLOGICAL MATERIALS
BIOLOGY
BLOOD
BODY
BODY FLUIDS
EQUIPMENT
LASERS
LIQUID LASERS
MAMMALS
MATERIALS
MEMBRANE PROTEINS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
ORGANS
PROTEINS
RADIATION EFFECTS
RADIATIONS
SOMATIC CELLS
THYLAKOID MEMBRANE PROTEINS
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560152* - Radiation Effects on Animals- Animals