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Axial super-resolution double-photon fluorescence microscopic device and method

A two-photon fluorescence and super-resolution technology, used in microscopes, measurement devices, fluorescence/phosphorescence, etc., can solve the problems of unfavorable axial information of tissue samples, and the axial resolution of the system is not improved, and achieves simple structure and imaging. deep effect

Active Publication Date: 2019-08-20
ZHEJIANG UNIV
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

[0006] However, the axial resolution of the system is still not improved, which is not good for analyzing the axial information of tissue samples

Method used

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  • Axial super-resolution double-photon fluorescence microscopic device and method
  • Axial super-resolution double-photon fluorescence microscopic device and method
  • Axial super-resolution double-photon fluorescence microscopic device and method

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Embodiment

[0038] see figure 1 , the axial super-resolution two-photon fluorescence microscopy device of the present embodiment includes a femtosecond laser 1, an optical parametric oscillator 2, a first half-wave plate 3, a polarizing beam splitter 4, a mirror 5, and a mirror 6, Polarizer 7, spatial light modulator 8, mirror 9, mirror 10, second half-wave plate 11, quarter-wave plate 12, mirror 13, mirror 14, scanning mirror 15, scanning mirror 16, Scanning lens 17 , field lens 18 , mirror 19 , dichroic mirror 20 , microscope objective 21 , sample 22 , optical filter 23 , collecting lens 24 , photomultiplier tube 25 and computer 26 .

[0039] use figure 1 The axial super-resolution two-photon microscopy method realized by the device shown is as follows:

[0040] (1) Femtosecond laser 1 emits illumination light, exits through optical parametric oscillator 2, passes through first half-wave plate 3 to become s-light, p-light ratio modulated polarized light, passes through first half-wave...

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Abstract

The present invention discloses an axial super-resolution double-photon fluorescence microscopic device and method, relates to the field of laser spot scanning microscopy, and realizes double-photon axial super-resolution imaging of a sample. The method emits an ultrashort pulse laser through a femtosecond laser to achieve two-photon excitation of the fluorescent sample; and performs phase modulation to the excitation light through a spatial light modulator, sequentially scans the sample with an axial solid spot and an axial hollow spot, and performs weight difference to two scanned two-photonimages obtained to finally obtain an axial super-resolution image. Compared with other two-photon axial super-resolution imaging microscopes, the device has a simple structure and a large imaging depth, and provides a good research method for life science and nanotechnology.

Description

technical field [0001] The invention relates to the field of laser point scanning microscopy, in particular to an axial super-resolution two-photon fluorescence microscopy device and method. Background technique [0002] Two-photon microscopy is increasingly used in life sciences. Compared with traditional single-photon fluorescence microscopy, two-photon can achieve a deeper penetration depth, and because two-photon excitation requires a higher photon density, the absorption of two-photon is only limited to a small range near the focal point, The axial resolution is improved to a certain extent. [0003] In addition, two-photon imaging usually uses an excitation light source in the infrared band, which causes less photodamage to living cells and tissues, and can perform long-term imaging. Although calculated according to Abbe's diffraction theory, the longer the wavelength, the greater the diffraction limit, but two-photon is widely used in biological research due to its ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64G02B21/00G02B21/34
CPCG01N21/6458G01N2021/6463G02B21/0068G02B21/0076G02B21/34
Inventor 匡翠方何敏菲孙试翼张智敏王文生杨欣刘旭张克奇毛磊
Owner ZHEJIANG UNIV
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