2020 Fiscal Year Annual Research Report
Investigation of mechanisms of cholesterol induced NETs by Raman Microscopy
Project/Area Number |
19K23865
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Research Institution | Osaka University |
Principal Investigator |
LELLIOTT PATRICK 大阪大学, 免疫学フロンティア研究センター, 特任研究員 (50844065)
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Project Period (FY) |
2019-08-30 – 2021-03-31
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Keywords | Neutrophil / NET / Cholesterol / Raman / Spectroscopy |
Outline of Annual Research Achievements |
- Neutrophil extracellular traps (NETs) were measured by Raman and quantitative phase microscopy coupled with fluorescence measurement to distinguish non-responding and necrotic cells. - An analysis pipeline was optimized for PCA and machine learning in order to classify NETs according to their stimulant and identify signature molecular differences in the cells response which were detected by Raman. - Cholesterol, cholesterol crystals, and oxidized LDL were tested for the ability to stimulate NET formation under different doses, times, and conditions. - In order to make results more applicable to in vivo conditions, an approach for the formation and measurement of NETs in whole blood was developed and validated using model NET stimuli.
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