水溶性菁染料是性能优良的荧光标记染料, 具有很好的水溶性和很高的摩尔吸光系数，其琥珀酰亚胺酯是最常用的脂肪氨基标记试剂，广泛用于蛋白、抗体、核酸及其他生物分子的标记和检测。特别是Cy5.5和Cy7的吸收在近红外区，已广泛用于活体成像领域来代替放射性元素。在哺乳动物体内血红蛋白是吸收可见光的主要成分, 能吸收中蓝绿光波段的大部分可见光; 水和脂质主要吸收红外线, 因此选择激发和发射光谱位于650nm-900nm(近红外窗口)的近红外荧光标记的红光虽然有部分散射消耗但大部分可以穿透哺乳动物组织被敏感的CCD检测到，更有利于活体光学成像，特别是深层组织的荧光成像。
Solvent：Water or DMSO
SPF级BALB/C裸鼠，6-8 周龄, 18-20克, 实验前24 h 自由进食、饮水。
实验裸鼠麻醉后，将裸鼠俯卧位平放于小动物多光谱活体成像系统的记录暗箱中。实验时将Cy7或Cy7标记的生物分子或药物DMSO稀释后,于裸鼠尾静脉注射200μL( 0.5 mg/g) [最佳用量和时间需要客户根据自己的仪器和药物试剂等条件优化],每5min 记录1 张动物在体内发射荧光的成像图片,分析荧光药物的分布情况。对照鼠不注射药物，进行同时记录。记录结束后迅速解剖裸鼠的心、肝、脾、肺、肾等脏器，进行成像。
Cy7 检测时激发波长700～770 nm带通,发射波长790 nm长通。液晶可调谐滤光片扫描范围780～950 nm ,扫描步进10 nm。曝光时间为500 ms。
未开封的粉末在避光干燥-20℃存放12月。CyDye NHS水溶液现配现用不能储存。任何溶解后的CyDye NHS粉末最好立即使用；无水的DMSO溶液-20°C保存最多2个星期。
Water soluble cyanine dye is a kind of fluorescent labeling dye with excellent performance. It has good water solubility and high molar absorptivity. Its succinimide ester is the most commonly used fatty amino labeling reagent, which is widely used for the labeling and detection of proteins, antibodies, nucleic acids and other biological molecules. In particular, the absorption of cy5.5 and cy7 in the near infrared region has been widely used in the field of in vivo imaging to replace radioactive elements. In mammals, hemoglobin is the main component of absorption of visible light, which can absorb most visible light in the blue-green band; water and lipid mainly absorb infrared light, Therefore, although the red light with excitation and emission spectrum located at 650 nm-900 nm (near-infrared window) has some scattering consumption, most of the red light can penetrate mammalian tissues and be detected by sensitive CCD, which is more conducive to in vivo optical imaging, especially fluorescence imaging of deep tissue.
Solvent：Water or DMSO
Living imaging of nude mice
SPF BALB / C nude mice, 6-8 weeks old, 18-20g, were free to eat and drink 24 hours before the experiment.
After anesthetized, the nude mice were placed in prone position in the recording box of small animal multispectral imaging system. In the experiment, cy7 or cy7 labeled biomolecule or drug DMSO was diluted, and then 200 μ L (0.5 mg / g) was injected into the tail vein of nude mice [the optimal dosage and time need to be optimized according to the conditions of their own instruments and drug reagents], and 1 imaging picture of the animal emission in vivo was recorded every 5 minutes to analyze the distribution of fluorescent drugs. The control rats were not injected with drugs, and were recorded at the same time. After recording, the heart, liver, spleen, lung, kidney and other organs of nude mice were dissected and imaged.
The excitation wavelength is 700-770 nm and the emission wavelength is 790 nm. The scanning range of liquid crystal tunable filter is 780-950 nm and the scanning step is 10 nm. The exposure time was 500 ms.
Different drug metabolism time is different. When injected into nude mice, fluorescence immediately distributes throughout the body, and then gradually accumulates to the bladder, showing the characteristics of significant renal excretion, generally 4-6 hours, as fast as 30 minutes; if it is cy7 labeled drugs targeted at bone and other parts, some customers report that fluorescence imaging can still be detected in vivo imaging system one week later.
Observation of organ sections: the dissected organs were quickly placed in 4% paraformaldehyde fixed for more than 4 hours, 0%, 20%, 30% PBS sucrose were successively sunk to the bottom, 20 μ m slices were sliced, the slides were pasted with polylysine acid, dried and DAPI staining. The laser is 633 nm He Ne.
Stability and storage
The unsealed powder was dried away from light and stored at – 20 ℃ for 12 months. CyDye NHS aqueous solution can not be stored when it is prepared and used. Any dissolved CyDye NHS powder should be used immediately; the anhydrous DMSO solution should be stored at – 20 ° C for up to 2 weeks.