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Two-photon excitation delay detection fluorescence imaging analysis method and equipment for live animal

An analysis equipment and fluorescence imaging technology, applied in the field of in vivo fluorescence image analysis of animals, can solve problems such as difficult application of high-efficiency two-photon excitation imaging analysis

Active Publication Date: 2019-01-04
PEKING UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • 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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  • Two-photon excitation delay detection fluorescence imaging analysis method and equipment for live animal
  • Two-photon excitation delay detection fluorescence imaging analysis method and equipment for live animal
  • Two-photon excitation delay detection fluorescence imaging analysis method and equipment for live animal

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] Example 1. Analysis of tumor targeting kinetics of fluorescent nanoprobes in tumor-bearing mice

[0071] 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 bpt's styrene-maleic anhydride copolymer (SMA) nanoparticles (16nm), obtained two kinds of two-photon sensitized Eu with different tumor recognition molecules on the surface 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 move significantly 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 exci...

Embodiment 2

[0075] Example 2. Comparison of two-photon excitation time-lapse fluorescence detection imaging and ordinary two-photon excitation fluorescence imaging in live animal imaging analysis

[0076] 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

[0077] Embodiment 3, the detection depth experiment of the imaging analysis method of the present invention

[0078] 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 delay detection fluorescence imaging analysis method and equipment for a live animal. The method is used for analyzing the time-varying kinetic propertyof distribution, fluorescence intensity or a probe concentration of nano fluorescence probes (or fluorescence nano drug carrier simulated probes) in a tissue and an organ in an animal body; the in-vivo fluorescence nano probes are subjected to two-photon excitation by red light or near infrared pulse laser, so as to perform delay detection on the fluorescence intensity and the time-varying kinetic property of an in-vivo specific part by a fluorescence imaging mode; the method has the advantages of large imaging analysis depth, high reliability, high detection sensitivity and the like.

Description

[0001] This application is a divisional application of a Chinese invention patent application with an application date of February 4, 2016, application number 201610078236.6, and a Chinese invention patent application titled "Fluorescence Imaging Analysis Method and Equipment for Two-photon Excitation Delayed Detection of Live Animals"; and the application date is required On February 16, 2015, the application number is 201510085138.0, and the priority of the Chinese invention patent application titled "Two-photon excitation time-lapse detection fluorescence imaging analysis method and equipment for living animals". technical field [0002] 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 applicati...

Claims

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Application Information

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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
Owner PEKING UNIV
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