1992 Fiscal Year Final Research Report Summary
Development of an automatic high-sensitive assay system for cells producing a specific antibody or a cytokine.
| Project/Area Number |
02557108
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| Research Category |
Grant-in-Aid for Developmental Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Laboratory medicine
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| Research Institution | Osaka University |
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Principal Investigator |
IWATANI Yoshinori Osaka University,Medical School,Associate Professor, 医学部, 助教授 (60168581)
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| Co-Investigator(Kenkyū-buntansha) |
SUZUKI Hajime Olympus Opticus, 2nd Dept.of Development, Chief, 第2開発部, 主任
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| Project Period (FY) |
1990 – 1992
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| Keywords | Specific antibody-producing cells / Cytokine-producing cells / ELISPOT assay / Automation / Standardization |
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| Research Abstract |
We tried to develop a high-sensitive assay for detection of cells producing a specific antibody or a cytokine with the automatic counting system. First, we developed an ELISPOT assay for cells producing thyroid-specific autoantibodies. The sensitivity of this assay was higher than that of a radioimmunoassay in terms of the detection of thyroid-autoantibody production. Second, we developed an automatic counting system for spots formed in the bottom of each well in an ELISPOT assay using an analyzer for color pictures. In this system, a scene of the bottom of each well including spots magnified in a metal microscope, and the magnified scene was shot by a color TV camera and analyzed by high-performance analyzer for color pictures. Each well of a plate was analyzed in turn by moving the plate on the metal microscope with a well scanner. In the high-performance analyzer, only spots with given ranges of size and depth were counted, but the counting number of spots depended on the degree of color development in the well. Therefore, we subtracted the degree of color development in a control well from that in a test well. Using this system, we found a significant correlation between the spot numbers counted by the analyzer and by manual counting. Furthermore, this system can be used for ELISPOT assays for cells producing various immunoglobulin classes of thyroid autoantibodies, but it was difficult to count the pale spots formed in a ELISA assay for cells producing IL-2 or IFN-gamma.
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