小动物活体成像

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
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
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
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
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
+ D-luciferin substrate
Fluorescent Proteins
Quantum dots and Nanoparticles
+ excitation light source
Transfection Transfetin labeling
Direct cell/protein labeling Direct cell/protein
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
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
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
ห้องสมุดไป่ตู้
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
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
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
Time point 3
Time point 4
PE Methodology = the same 6 animals over four treatment points
Time point 1
Time point 2
Time point 3
Time point 4
光学成像 Optical imaging
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
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
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。
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)