1988 Fiscal Year Final Research Report Summary
Development of chromosome sorting techniques using a dual laser chromosome sorter
| Project/Area Number |
61880023
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| Research Category |
Grant-in-Aid for Developmental Scientific Research
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| Allocation Type | Single-year Grants |
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| Research Field |
分子遺伝学・分子生理学
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| Research Institution | Osaka University |
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Principal Investigator |
MATSUBARA Kenichi Osaka University,Professor, 細胞工学センター, 教授 (20037394)
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| Co-Investigator(Kenkyū-buntansha) |
OCHIYA Takahiro Osaka University,Research Associate, 細胞工学センター, 助手 (60192530)
FUKUSHIGE Shinichi Osaka University,Research Associate, 細胞工学センター, 助手 (90192723)
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| Project Period (FY) |
1986 – 1988
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| Keywords | Cell Sorter / Dual Laser / 染色体 |
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| Research Abstract |
We have developed chromosome sorting techniques using a dual laser chromosome sorter to analyze the structure of human chromosomes.
Human chromosomes stained with Hoechst 33258 (Ho) and chromomycin A3 (CA3) flowed sequentially through two laser beams, one adjusted to 457.9nm to excite CA3 and the other adjusted to 351.1-363.8nm to excite Ho. The Ho and CA3 contents of individual chromosomes were determined by measuring the intensities of fluorescence that were emitted as they passed two laser beams. These measurements were accumulated to form fluorescence intensity distributions.
Sorting techniques have evolved from the ability to sort 10-20 human chromosome fractions with a single laser system to sorting 21 fractions including 15 unique types with a dual laser system.
dFurthermore, chromosome types that cannot be sorted at high purity from chromosomes isolated from human cells can be sorted from human-hamster hybrid cell lines containing one or a few human chromosomes.
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