Utility of frozen platelets for a platelet antibody assay using flow cytometric analysis. 1988

J Lazarchick, and P C Das, and T J Jones, and R J Russell, and S A Hall
Department of Pathology, Medical University of South Carolina, Charleston 29425.

The immunoreactivity of a PLA1 antibody-containing serum to frozen/thawed platelets prepared by four different procedures was measured to assess the practicality of preparing a platelet reagent that could be easily stored and readily used in an immunofluorescent platelet antibody detection system. These frozen aliquots were analyzed at intervals up to 23 weeks after initial freezing and storage. For analysis, the frozen platelets were thawed, washed, resuspended to 2.5 x 10(5) platelets/mL and incubated with a dilution of fresh autologous serum that lacked platelet antibody or with a similar dilution of frozen human serum containing IgG anti-PLA1 antibody. At each interval, a freshly drawn platelet sample from the same donor was incubated in the same manner and used for comparison. All platelet mixtures were then washed and incubated with fluorescein conjugated goat F (ab')2 antihuman IgG. After repeat washing, each mixture was then analyzed with a flow cytometer for the extent of fluorescent antibody bound to each platelet mixture. Anti-PLA1 antibody reactivity with either the fresh or the frozen/thawed platelets remained stable over the period of analysis, with no significant difference in immunofluorescence with either fresh or frozen platelets as target cells. Platelet recovery following the thawing step ranged from 12% to 36% and was independent of the storage time. These studies suggest that frozen platelets can be readily used as reagents for platelet antibody assays.

UI MeSH Term Description Entries
D008297 Male Males
D001792 Blood Platelets Non-nucleated disk-shaped cells formed in the megakaryocyte and found in the blood of all mammals. They are mainly involved in blood coagulation. Platelets,Thrombocytes,Blood Platelet,Platelet,Platelet, Blood,Platelets, Blood,Thrombocyte
D001793 Blood Preservation The process by which blood or its components are kept viable outside of the organism from which they are derived (i.e., kept from decay by means of a chemical agent, cooling, or a fluid substitute that mimics the natural state within the organism). Blood Preservations,Preservation, Blood,Preservations, Blood
D005434 Flow Cytometry Technique using an instrument system for making, processing, and displaying one or more measurements on individual cells obtained from a cell suspension. Cells are usually stained with one or more fluorescent dyes specific to cell components of interest, e.g., DNA, and fluorescence of each cell is measured as it rapidly transverses the excitation beam (laser or mercury arc lamp). Fluorescence provides a quantitative measure of various biochemical and biophysical properties of the cell, as well as a basis for cell sorting. Other measurable optical parameters include light absorption and light scattering, the latter being applicable to the measurement of cell size, shape, density, granularity, and stain uptake. Cytofluorometry, Flow,Cytometry, Flow,Flow Microfluorimetry,Fluorescence-Activated Cell Sorting,Microfluorometry, Flow,Cell Sorting, Fluorescence-Activated,Cell Sortings, Fluorescence-Activated,Cytofluorometries, Flow,Cytometries, Flow,Flow Cytofluorometries,Flow Cytofluorometry,Flow Cytometries,Flow Microfluorometries,Flow Microfluorometry,Fluorescence Activated Cell Sorting,Fluorescence-Activated Cell Sortings,Microfluorimetry, Flow,Microfluorometries, Flow,Sorting, Fluorescence-Activated Cell,Sortings, Fluorescence-Activated Cell
D005615 Freezing Liquids transforming into solids by the removal of heat. Melting
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000906 Antibodies Immunoglobulin molecules having a specific amino acid sequence by virtue of which they interact only with the ANTIGEN (or a very similar shape) that induced their synthesis in cells of the lymphoid series (especially PLASMA CELLS).

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