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Three-dimensional imaging system and method in deep scattering medium

A three-dimensional imaging and medium technology, applied in the direction of material excitation analysis, fluorescence/phosphorescence, etc., can solve the problems of phosphor imaging and low efficiency of three-dimensional images

Active Publication Date: 2013-02-27
BEIJING LUSTER LIGHTTECH
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Problems solved by technology

[0005] In order to solve the technical problem that the existing 3D technology uses point-by-point scanning to acquire 3D images inefficiently and cannot image phosphors in scattering media, the present invention provides a 3D imaging system and method in deep scattering media. The invention proposes to use line-focused beams instead of point-focused beams in ordinary scanning fluorescence microscopy imaging technology to realize three-dimensional imaging in scattering media

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  • Three-dimensional imaging system and method in deep scattering medium
  • Three-dimensional imaging system and method in deep scattering medium
  • Three-dimensional imaging system and method in deep scattering medium

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[0050] In order to efficiently excite the two-photon fluorescence in the fluorescently labeled sample and scan the sample at high speed, the excitation light source generally adopts an ultrashort pulse mode-locked laser with a high repetition rate (such as 80MHz). For example, the Mai Tai ultrafast femtosecond laser from Sepctra-Physics can be used, which has a wide spectral adjustment range (690nm~1040nm), and can excite common fluorescent markers to produce two-photon fluorescence. The pulse width is generally selected between 100 and 200 femtoseconds, and a larger pulse width will reduce the two-photon excitation efficiency, thereby weakening the excitation depth of the Bessel beam.

[0051] The microscope objective serves both as a component for scanning Bessel beams and as a component for reverse fluorescence collection in the present invention. In order to enable the scanning field to have a better Bessel focusing field at different scanning angles, it is advisable to us...

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Abstract

A three-dimensional imaging system in a deep scattering medium includes a light source, a Bessel light beam generating and scanning device, a fluorescence collection system, a signal detection component, and an image acquisition system which are arranged in order along the optical path propagation direction, wherein the Bessel light beam generating and scanning device includes a translation stage, a cone mirror, a first lens, a galvanometer and a 4f system which are arranged along the optical path. The system and method help to solve the technical problems of inefficiency that conventional three-dimensional imaging technology uses point by point scanning to obtain a three-dimensional image and the phosphor in the scattering medium can not be imaged, use standard Bessel light beam which can generate a line focusing along light propagation direction as excitation light to replace conventional Gaussian beam which produces point focusing in the two-photon excitation fluorescence scanning microtechnic, and change original flat scaning into volume scanning to achieve quick acquisition the internal fluorescence distribution images in the scattering medium.

Description

technical field [0001] The invention proposes a system and method for performing three-dimensional imaging in a deep scattering medium. Two-photon fluorescence is generated by using ultrashort pulse line focused beam excitation to realize three-dimensional spatial distribution imaging of fluorescent markers in scattering media, especially inside biological tissues. The invention is suitable for research and application in the fields of biology, medicine, biophysics and environmental science. Background technique [0002] Fluorescence microscopy has become an essential device in biomedical research due to its high sensitivity, high spatial resolution, and rich molecular contrast mechanisms. Due to the light scattering effect, ordinary fluorescence microscopes can only image very thin (<10 micron) samples or a very thin layer on the surface of the sample in order to obtain ideal high-resolution images. For the microscopic imaging of phosphors in scattering media, confocal...

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Application Information

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IPC IPC(8): G01N21/64
Inventor 叶彤杨延龙姚保利雷铭郑娟娟严绍辉但旦
Owner BEIJING LUSTER LIGHTTECH
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