| Project/Area Number |
26860252
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Multi-year Fund |
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| Research Field |
Human pathology
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| Research Institution | Tokushima Bunri University |
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Principal Investigator |
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| Project Period (FY) |
2014-04-01 – 2016-03-31
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| Project Status |
Completed (Fiscal Year 2015)
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| Budget Amount *help |
¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Fiscal Year 2015: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
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| Keywords | Laser Microdissection / LCM / 分子病理学 / 免疫組織化学 / 遺伝子発現解析 / リアルタイムPCR |
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| Outline of Final Research Achievements |
In this work, I developed a new method for combined immunohistochemical and molecular biological analyses at the cellular-scale by laser capture microdissection (LCM) using formalin- or paraformaldehyde (PFA)-fixed tissues that were routinely used in histopathological examinations in clinical medicine.
I optimized immunohistochemical method for LCM and molecular biological analysis using tissue sections coated with a water-soluble polymer that improved cell identification by inhibiting cell shrinkage during drying process of sections. I also established an efficient RNA extraction method that was sequential treatment of isolated specimens by LCM with de-crosslinking, elimination of genomic DNA and extraction of RNA in the same one-tube. These approaches made it possible to detect expressed genes (mRNAs) from 5 cells isolated from PFA-fixed sections of mouse brain by LCM.
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