Genetic Marker
Label Cells Label Bacteria
Label Proteins
超高灵敏度CCD—最低的温度最少的噪 音
-90℃
-50℃
-30℃
超高灵敏度CCD—最高的量子效率
量子效率(Quantum Efficiency)—— 发射出的光子数量与检测器 光子数的比值，用以表诉CCD在不同波长下的响应值
Optical imaging showed Listeria can replicate in the murine gall bladder
感染性疾病
Effects of Antibiotics on S. aureus Biofilm Infections
Day 7 Untreated Control Day 9 Day 12 Day 16 Day 22
CCD Optics
• 透过灵敏的光学元件及CCD， 可将光学信号转成电子信号， 再转换成图像输出。
Bioluminescent Source
Imaging Basics – Reporter Molecules
Luciferase
Fluorescent dyes
+ ATP and O2 – Live cells
Precision filters
Low f-number and large diameter lens
Single cell (flux: 460528 photons/sec)
Gives high sensitivity and uniform light collection
Rabinovich et al, PNAS,2008
中草药抑制癌症 研究
粉防己碱 — TET 抑制人类结直肠癌 HCT116 — 人类结直肠癌细胞系
中医方向研究
刮痧的过程上调血红素加氧酶。 血红素加氧酶对细胞起到保护作 用，能对抗氧化压力
天津医科大学肿瘤医院 公共实验室 杨毅
从体外到体内-从实验室到临床
从侵入式到非侵入式动物实验
长时间观测 降低个体间差异 减少动物用量 Traditional Methodology = 24 animals over four treatment points
Time point 1
Time point 2
检测残余病灶
药物研究
Luc标记的前列腺肿瘤细胞株皮下模型
3 weeks of vehicle treatment
3 weeks of paclitaxel treatment a false cure by palpation
感染性疾病
No Additional Substrate Needed
5 6
medium
Radiance: Photons per second
整体解决方案
标记 技术
荧光素酶基因标记的 细胞、细菌和转基因动物
成像 检测
高灵敏度的可见光检测设备：
定量 分析
定量分析方案
Non-Invasive Detection of Micrometastases
癌症研究
5×105 4T1-luc2-1A4 cells orthotopically into the abdominal mammary fat pads
Kim et al, pLOs One et al, 2010
In vivo imaging of s.c. implanted T cells transduced with optimized firefly luciferase (left) and a ‘single’ 4T1 breast cancer cell (right)
microPET microCT
microSPECT
Optical bioluminescence
Optical fluorescence
microMRI
光学成像的优势
1.高通量 2.无辐射问题
3.操作简易
4. 功能性成像 5. 多模式成像 (生物发光 及 荧光 )
光学成像原理-活体成像
• 体内光源发出的光，经过散射 吸收後到达表面形成光斑。
Rifampin 25mg/kg
Ciprofloxacin 15 mg/kg
Tobramycin 20 mg/kg
干细胞研究
Co-registration of IVIS data with other modalities (e.g. PET) offers complementary insights into biological processes
精诺真的绝对定量功能-原始数据
Raw Signal (Counts)
Exp time: sec Binning: Day:
1600
30 sec small 1
30 sec small 2 3
60 sec small 4
60 sec small
60 sec medium 5
60
medium 6
Peak Counts
Time point 3
Time point 4
PE Methodology = the same 6 animals over four treatment points
Time point 1
Байду номын сангаас
Time point 2
Time point 3
Time point 4
光学成像 Optical imaging
1200 800 400
精诺真的绝对定量功能-绝对定量
Calibrated Signal (Photons per second)
Exp time:
30 sec
30 sec
60 sec
60 sec
60 sec
60 sec
Binning:
Day:
small
1 2
small
3
small
4
small medium
+ D-luciferin substrate
Fluorescent Proteins
Quantum dots and Nanoparticles
+ excitation light source
Transfection Transfetin labeling
Direct cell/protein labeling Direct cell/protein
Wavelength (nm)
超高灵敏度CCD-最高的体内检测灵 敏度
Cooled (absolute -90C) camera with large CCD chip area for high sensitivity light detection Most sensitive system available Resolves multiple bioluminescent reporters Detects down to even a single cell in vivo
BLI (top): ES cell viability and proliferation (biological quantitation)
PET (bottom): resolved migratory patterns and clinical translatability
干细胞研究
Bioluminescence Imaging of Neuronal Stem Cell Migration
100 90 80
Qunatum Efficiency (%)
70 60 50 40 30 20 10 0 400 500 600 700 800 900
在同一波长下QE值越高 CCD品质越好
CCD对于不同波长的光的 响 应时间的敏感度不同
背照式CCD比前照式CCD 有更好的量子效率
EEV 47-10 Kodak 2001