Live animal two-photon excitation time-lapse detection fluorescence imaging analysis method and equipment

An analysis method, a technique of fluorescence imaging, which is applied in the field of in vivo fluorescence image analysis of animals, and can solve problems such as difficult application of high-efficiency two-photon excitation imaging analysis

Active Publication Date: 2019-07-05
PEKING UNIV +1
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

This technology uses point-by-point scanning imaging, so it is not suitable for live animal imaging under large field of view
At the same time, due to the point-by-point scanning working mode, under the limitation of imaging time, the working time of each point must be on the order of nanoseconds, which makes it difficult to apply this kind of technology to the efficient two-photon excitation imaging analysis of living animals

Method used

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  • Live animal two-photon excitation time-lapse detection fluorescence imaging analysis method and equipment
  • Live animal two-photon excitation time-lapse detection fluorescence imaging analysis method and equipment
  • Live animal two-photon excitation time-lapse detection fluorescence imaging analysis method and equipment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Example 1. Analysis of tumor targeting kinetics of fluorescent nanoprobes in tumor-bearing mice

[0070] The HepG-2 liver cancer model for two-photon excitation time-resolved imaging was established on the right hind leg of nude mice. In the experiment, transferrin (Tf) and RGD modified with 10% Eu(tta) were used respectively. 3 Styrene-maleic anhydride copolymer (SMA) nanoparticles (16nm) of bpt obtained two photon-sensitized Eu 3+ Luminescent fluorescent nanoprobes. 300 microliters of colloid solution containing 1.5 mg of fluorescent nanoprobes was injected through the tail vein, and then the mice were anesthetized using an inhalation respiratory anesthesia system to ensure that the mice would not produce significant displacement during the imaging process. On the two-photon excitation time-resolved imaging device of the present invention, a femtosecond pulsed laser with a wavelength of 800nm ​​is used to excite the fluorescent nanoprobe in the body of the desired a...

Embodiment 2

[0074] Example 2. Comparison of two-photon excitation time-lapse fluorescence detection imaging and ordinary two-photon excitation fluorescence imaging in live animal imaging analysis

[0075] In this embodiment, the conditions of the two-photon excitation time-lapse detection imaging experiment are the same as those in Embodiment 1, and the excitation conditions are the same in the ordinary two-photon excitation fluorescence imaging experiment, and the time delay is zero nanoseconds. We performed two-photon excitation time-lapse fluorescence detection imaging and ordinary two-photon excitation fluorescence imaging on the tumor area of ​​tumor-bearing mice before and after fluorescent probe injection. The result is as Figure 8 shown. The experimental results show that using the method and device provided by the present invention can effectively eliminate the influence of background fluorescence on imaging analysis in two-photon excitation live animal imaging analysis, and ob...

Embodiment 3

[0076] Embodiment 3, the detection depth experiment of the imaging analysis method of the present invention

[0077] Fix the fluorescent nanoprobe on the bottom of the container, inject the adipose tissue simulation liquid into the container, adjust the thickness of the adipose tissue simulation liquid covered on the bottom probe, and use the time-lapse fluorescence detection method and equipment of the present invention to allow the excitation light to pass through the top The probe is excited by the tissue simulating fluid, and imaging analysis is performed above the liquid surface. The excitation and detection conditions are as in Example 1. Such as Figure 9 As shown, the results show that the detection depth of fluorescence imaging in this experiment is greater than 7mm, which is significantly improved compared with the previous reliable detection depth of fluorescence imaging. Studies have shown that the detection depth can be further greatly improved by changing the ex...

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Abstract

The invention discloses a two-photon excitation time-lapse detection fluorescence imaging analysis method and equipment for living animals, which are used for analyzing the distribution and fluorescence intensity of fluorescent nano-probes (or fluorescent nano-drug-carrying analog probes) in tissues and organs in animals. Or the kinetic properties of the probe concentration changing with time, using red light or near-infrared pulsed laser two-photon to excite fluorescent nanoprobes in vivo to emit light, and detecting the fluorescence intensity of specific parts in vivo and its dynamic properties with time by time-lapse detection fluorescence imaging , has the advantages of large imaging analysis depth, high reliability and high detection sensitivity.

Description

technical field [0001] The present invention relates to animal in vivo fluorescence image analysis technology, in particular to a fluorescence imaging analysis method for the distribution and transport kinetic properties of nano-drug-loaded simulated probes or fluorescent nano-probes in living animals, and equipment for implementing the analysis, And the application of the method and the device in the characterization of the nano drug carrier tumor targeting kinetic properties. Background technique [0002] Animal in vivo fluorescence imaging technology has the characteristics of high sensitivity and small damage, and has developed into a common method for imaging living animals, and is widely used in the fields of life science, biomedicine and drug research and development [V.Wagner, A.Dullaart, A. Bock, A. Zweck, The emerging nanomedicine landscape. Nature Biotechnology. 2006, 24, 1211-1217.]. In vivo fluorescence imaging systems have broad application prospects in many r...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64A61B5/00
CPCA61B5/0071G01N21/6402G01N21/6456G01N21/6486
Inventor 王远付立民张建平杨文文学刘禹辰王川西
Owner PEKING UNIV
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