미토콘드리아의 pH를 검출하기에 적합한 이광자 형광 프로브의 합성
Synthesis of Benzochromene-Derived Two-Photon Fluorescence Probes for Mitochondrial pH
- 주제(키워드) TP Fluorescence Probes , Mitochondrial pH
- 발행기관 아주대학교
- 지도교수 김환명
- 발행년도 2015
- 학위수여년월 2015. 8
- 학위명 석사
- 학과 및 전공 일반대학원 에너지시스템학과
- 실제URI http://www.dcollection.net/handler/ajou/000000020201
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Two-photon microscopy (TPM) is a fluorescence imaging technique which provides greater tissue penetration, limits the excitation volume, and induces less photobleaching, phototoxicity and photodamage. Mitochondria play a crucial role in cell metabolic processes such as energy production, cell signaling, Ca2+ regulation, production in reactive oxygen spec ies and triggering of cell death.The mitochondrial function depends on its pH. Mitochondrial pH plays a significant role in driving force for the mitochondrial cation homeostasis, production of ATP and neurotransmission an insulin secretion. Here, we report two mitochondria-targetable two-photon fluorescent pH sensitive probes (CMP-1 and CMP-2) allowing for a quantitative measurement of mitochondrial pH in living cells and tissues. In the probe architecture, we assimilated the donor OH group at the C-8 position of the benzochromene scaffold which enhance the dipole moment and offer larger intramolecular charge transfer (ICT) process. Benzochromene scaffolds of CMP-1 and CMP-2 are different. CMP-1 is bent-π-shaped and CMP-2 is linear-π-shaped. Because conjugation of CMP-2 is larger than CMP-1, the ICT process in CMP-2 would enhance and the pH monitoring with CMP-2 would be occurred in red region which is ruled out the autofluorescence and helpful for enhanced the spectral separation from cellular absorption in biological trial.
more목차
Ⅰ. General Introduction 1
1. Introduction of Two-Photon Microscopy 1
2. Introduction of Intramolecular Charge Transfer (ICT) 2
3. Introduction of mitochondrial pH 3
4. Design of Two-Photon Probes for Mitochondrial pH 4
Ⅱ. Synthesis of CMP-1 and CMP-2 6
1. Synthetic route of CMP-1 6
2. Synthetic route of CMP-2 10
Ⅲ. Photophysical Data 16
1. Spectroscopic Measurements 16
2. Water Solubility 16
3. Fluorescence pH Titrations and measurement of pKa vlues 19
4. Measurement of Two-Photon Absorption (TPA) Cross Section (δ) 21
IV. Two-Photon Microscopy Bioimaging with CMP-1 and CMP-2 23
1. Photostability and Cytotoxicity 23
2. Co-localization experiments 24
3. Cell calibration 25
4. CCCP effect on mitochondrial pH 27
5. pH change inside the mitochondria upon nutrient deprivation 29
6. TPM images with CMP-2 in Live Brain Tissues 30
V. Conclusion 32
References 33
1H-NMR, 13C-NMR and HRMS spectrum of intermediates and CMP-1 36
1H-NMR, 13C-NMR and HRMS spectrum of intermediates and CMP-2 42
