(优选)荧光探针与分子传感器
合集下载
相关主题
- 1、下载文档前请自行甄别文档内容的完整性，平台不提供额外的编辑、内容补充、找答案等附加服务。
- 2、"仅部分预览"的文档,不可在线预览部分如存在完整性等问题,可反馈申请退款(可完整预览的文档不适用该条件!)。
- 3、如文档侵犯您的权益，请联系客服反馈,我们会尽快为您处理(人工客服工作时间：9:00-18:30)。
Stoke’s shift
Fluorescence lifetime 荧光寿命 Fluorescence quantum yield 荧光量子产率
Fluorescence Anisotropy 各项异性 r Fluorescence Polarization 荧光偏振 p
20050705
7
Excitation and emission spectra
15
Sensor (5)
竞争型
+
S-R
T
催化型
+
S-R
T
20050705
+
hv
催化剂中间体
+
16
Tracer The Fusion of Integal Membrane Protein
荧光素标记 鼠蛋白质
膜 鼠细胞
内
蛋
白
侧
向
运
37°C
动
融合
罗丹明标记 人蛋白质 人细胞
异核体
15°C
20050705
20050705
www. probes. co5 m
Jablonski diagram
S2
Vibrational relax, VR, 10-1210-14 s
S1
LUMO
Absorption 10-15 s
hA hA
S0
HOMO
Internal conversion, IC, 10-1110-13 s
Excitation spectrum em
F
Emission spectra
ex
F
1
ex
20050705
em
8
Fluorescence lifetime
Simplified Jablonski diagram
S1
hvA S0
Definition
S1 hvF
relaxation (10-12 s)
Application
ห้องสมุดไป่ตู้
1. Radical probes 2. Ion probes 3. NIRF Fluorescence image 4. DNA sequence
20050705
5. Protein probes 6. DNA probes 7. Membrane probes 8. pH probes
Intersystem crossing, ISC, 10-210-6 s
T1 Fluorescence
IC
hF 10-8 s
Phosphorescence ISC
10-3100 s hP
3210
Characteristics of Fluorophore
Excitation wavelength 激发波长 ex Emission wavelength 发射波长 em Extinction (absorption) coefficient 吸光系数
2
Fluorescence research
Key word: Fluorescen*
Period: Jan/2005-Jun/ 2005
Data base:
➢ 美国化学会 ( ACS )数据库 ➢ WPS电子期刊全文数据库 ➢ SDOS全文数据库（上交 镜像） ➢ 英国皇家化学会（RSC) 数据库
Hg2+
hv
20050E7.05M. Nolan, et al., J. Am. Chem. Soc., 2003, 125: 14270-14271
12
Sensor (3)
Fluorescence Resonance Energy Transfer
hv
FRET S
R
T
+
hv
hv’
hv
D,exD,emA,ex A,em F
1
Γ knr
Fluorescence Quantum Yield
Definition
The fluorescence quantum yield is the ratio of the number of photons emitted to the number absorbed
Expression
From Instant notes, Biochemistry, E2 17
Cell tracer
C7000 C68H105Cl2N3O
荧光染色的斑马鱼胚胎
A 30 µm–thick section of a zebrafish embryo stained with CellTracker CM-DiI (C7000, C7001) prior to immunohistochemical analysis. A one-dayold zebrafish embryo was immobilized and impaled in the hindbrain with a microelectrode filled with 1 mg/mL CellTracker CM-DiI in 95% ethanol. One day later, the brain was fixed, embedded, frozen in liquid nitrogen and sectioned on a cryostat. After blocking with 0.1% Triton X100 in PBS containing 2% BSA and 2% normal goat serum, the 30 µm–thick section was incubated with primary anti-glial antibody in conjunction with fluorescein goat anti–mouse IgG antibody. This section was then viewed sequentially through optical filter sets appropriate for rhodamine and fluorescein, and the resulting images were superimposed. The image was contributed by William Trevarrow, Beckman Institute, California Institute of Technology.
: Emission rate
knr Knr: nonradiative
decay rate
Expression
Average lifetime: the average time the molecule spends in the excited state prior to return to the ground state.
FLUORESCENCE SPECTROSCOPY
（优选）荧光探针与分子传感 器
XIAMEN UNIVERSITY
Fluorescent Probes
Introduction Fluorescent Sensor 荧光传感 Fluorescent Tracer 荧光示踪 FRET 共振能量转移荧光
Φ Γ Γ knr
Sensor (1)
S 信号单元， R 识别单元， T 靶分子
S
R
T
+
自由基传感
Z.-Y. Bian, X.-Q. Guo*, Analytical Science, 21 553-559, 2005
Sensor (2)
hu hv e - R S
D
T hv
hv
D
PET Photoinduced Electron Transfer
D,exD,emA,ex A,em F
Probing DNA
Molecular beacons 分子信标
F
Q
Target DNA
ssDNA binding protein
Q
Denaturing reagent
QF AD
20050705
14
Sensor (4)
+
S-R
T
hv hv
Al3+
hv
20050705
3566 篇 206 篇 1890 篇 120 篇
研究与应用领域涉及：生命 环境 材料 等
讲座要求了解： 荧光 探针 表征 应用 研究意义
20050705
3
Molecular Probe
Molecular reporter
20050705
From J. Fluorescence
4
Molecuhloamr perpoabgees
Fluorescence lifetime 荧光寿命 Fluorescence quantum yield 荧光量子产率
Fluorescence Anisotropy 各项异性 r Fluorescence Polarization 荧光偏振 p
20050705
7
Excitation and emission spectra
15
Sensor (5)
竞争型
+
S-R
T
催化型
+
S-R
T
20050705
+
hv
催化剂中间体
+
16
Tracer The Fusion of Integal Membrane Protein
荧光素标记 鼠蛋白质
膜 鼠细胞
内
蛋
白
侧
向
运
37°C
动
融合
罗丹明标记 人蛋白质 人细胞
异核体
15°C
20050705
20050705
www. probes. co5 m
Jablonski diagram
S2
Vibrational relax, VR, 10-1210-14 s
S1
LUMO
Absorption 10-15 s
hA hA
S0
HOMO
Internal conversion, IC, 10-1110-13 s
Excitation spectrum em
F
Emission spectra
ex
F
1
ex
20050705
em
8
Fluorescence lifetime
Simplified Jablonski diagram
S1
hvA S0
Definition
S1 hvF
relaxation (10-12 s)
Application
ห้องสมุดไป่ตู้
1. Radical probes 2. Ion probes 3. NIRF Fluorescence image 4. DNA sequence
20050705
5. Protein probes 6. DNA probes 7. Membrane probes 8. pH probes
Intersystem crossing, ISC, 10-210-6 s
T1 Fluorescence
IC
hF 10-8 s
Phosphorescence ISC
10-3100 s hP
3210
Characteristics of Fluorophore
Excitation wavelength 激发波长 ex Emission wavelength 发射波长 em Extinction (absorption) coefficient 吸光系数
2
Fluorescence research
Key word: Fluorescen*
Period: Jan/2005-Jun/ 2005
Data base:
➢ 美国化学会 ( ACS )数据库 ➢ WPS电子期刊全文数据库 ➢ SDOS全文数据库（上交 镜像） ➢ 英国皇家化学会（RSC) 数据库
Hg2+
hv
20050E7.05M. Nolan, et al., J. Am. Chem. Soc., 2003, 125: 14270-14271
12
Sensor (3)
Fluorescence Resonance Energy Transfer
hv
FRET S
R
T
+
hv
hv’
hv
D,exD,emA,ex A,em F
1
Γ knr
Fluorescence Quantum Yield
Definition
The fluorescence quantum yield is the ratio of the number of photons emitted to the number absorbed
Expression
From Instant notes, Biochemistry, E2 17
Cell tracer
C7000 C68H105Cl2N3O
荧光染色的斑马鱼胚胎
A 30 µm–thick section of a zebrafish embryo stained with CellTracker CM-DiI (C7000, C7001) prior to immunohistochemical analysis. A one-dayold zebrafish embryo was immobilized and impaled in the hindbrain with a microelectrode filled with 1 mg/mL CellTracker CM-DiI in 95% ethanol. One day later, the brain was fixed, embedded, frozen in liquid nitrogen and sectioned on a cryostat. After blocking with 0.1% Triton X100 in PBS containing 2% BSA and 2% normal goat serum, the 30 µm–thick section was incubated with primary anti-glial antibody in conjunction with fluorescein goat anti–mouse IgG antibody. This section was then viewed sequentially through optical filter sets appropriate for rhodamine and fluorescein, and the resulting images were superimposed. The image was contributed by William Trevarrow, Beckman Institute, California Institute of Technology.
: Emission rate
knr Knr: nonradiative
decay rate
Expression
Average lifetime: the average time the molecule spends in the excited state prior to return to the ground state.
FLUORESCENCE SPECTROSCOPY
（优选）荧光探针与分子传感 器
XIAMEN UNIVERSITY
Fluorescent Probes
Introduction Fluorescent Sensor 荧光传感 Fluorescent Tracer 荧光示踪 FRET 共振能量转移荧光
Φ Γ Γ knr
Sensor (1)
S 信号单元， R 识别单元， T 靶分子
S
R
T
+
自由基传感
Z.-Y. Bian, X.-Q. Guo*, Analytical Science, 21 553-559, 2005
Sensor (2)
hu hv e - R S
D
T hv
hv
D
PET Photoinduced Electron Transfer
D,exD,emA,ex A,em F
Probing DNA
Molecular beacons 分子信标
F
Q
Target DNA
ssDNA binding protein
Q
Denaturing reagent
QF AD
20050705
14
Sensor (4)
+
S-R
T
hv hv
Al3+
hv
20050705
3566 篇 206 篇 1890 篇 120 篇
研究与应用领域涉及：生命 环境 材料 等
讲座要求了解： 荧光 探针 表征 应用 研究意义
20050705
3
Molecular Probe
Molecular reporter
20050705
From J. Fluorescence
4
Molecuhloamr perpoabgees