Resonance Raman spectroscopic study and fluorescence cell imaging using π-conjugated chromophore-based nanoparticles
- 주제(키워드) Raman , Cell imaging , Nanoparticles
- 발행기관 고려대학교 대학원
- 지도교수 심상희
- 발행년도 2019
- 학위수여년월 2019. 2
- 학위구분 석사
- 학과 대학원 화학과
- 세부전공 분석화학 전공
- 원문페이지 44 p
- 실제URI http://www.dcollection.net/handler/korea/000000083210
- UCI I804:11009-000000083210
- 본문언어 영어
초록/요약
Raman scattering is an emerging contrast mechanism for biological imaging due to its narrow spectral bandwidth. However, the sensitivity is many orders of magnitude lower than that of fluorescence. Fluorescence offers extremely high sensitivity, but suffers from broad absorption and emission spectra. Since Raman and fluorescence are complementary to each other, we developed conjugated polymer-based nanoparticles as an imaging probe for both Raman and fluorescence for cellular imaging. The Raman-active vibrating groups in the nanoparticles that are electronically resonant to the π-conjugation system produced highly enhanced Raman scattering signals. When the nanoparticles were irradiated with 532 nm laser near the absorption maximum of the π-conjugated nanoparticles, the nanoparticles produced highly enhanced Raman signal for the vibrational modes at 1200-1800 cm-1, while emitting high far-red fluorescence to the sensitivity level of detecting single particles. By conjugating the carboxylic groups on the nanoparticles and the amine groups on proteins such as avidins and antibodies, we labeled specific proteins in the cell with nanoparticles and performed molecular-specific cell imaging experiments. Imaging probe with both fluorescence and Raman-activity is a unique and powerful tool that combines high multiplexing of Raman and single-particle sensitivity of fluorescence.
more목차
Abstract
Contents
1. Introduction 1
2. Resonance Raman spectroscopic study of nanoparticles 6
2.1. Materials and Methods 7
2.1.1. Materials 7
2.1.2. Raman spectroscopy 10
2.2. Results and Discussion 11
2.3. Conclusion 19
3. Fluorescence cell imaging using nanoparticles 20
3.1. Materials and Methods 21
3.1.1. Cell culture 21
3.1.2. Conjugation reaction 21
3.1.3. Cell immunostaining 23
3.1.4. Spinning confocal fluorescence microscopy 24
3.2. Results and Discussion 24
3.3. Conclusion 28
Appendix 29
A1. Baseline correction using Matlab code 30
References 31
Acknowledgement 36
