Method and device of stimulated emission depletion (STED) microscopy based on tangential polarized light

A technology of stimulated emission loss and tangentially polarized light, which is applied in the field of super-resolution microscopy to achieve the effects of small light-emitting area, reduced bleaching, and avoiding damage to samples

Inactive Publication Date: 2012-09-12
ZHEJIANG UNIV
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[0002] According to Abbe's diffraction limit theory, the size of the spot formed by the beam after being focused by a general lens can be expressed as Where λ is the wavelength of the...

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  • Method and device of stimulated emission depletion (STED) microscopy based on tangential polarized light
  • Method and device of stimulated emission depletion (STED) microscopy based on tangential polarized light
  • Method and device of stimulated emission depletion (STED) microscopy based on tangential polarized light

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Embodiment Construction

[0035] Such as figure 1 As shown, a stimulated emission loss microscopy device based on tangentially polarized light includes: a first laser 1, a second laser 2, a first single-mode fiber 3, a second single-mode fiber 4, and a first collimator lens 5. The second collimating lens 6, the first tangential polarization converter 7, the second tangential polarization converter 8, the first dichroic prism 9, the first phase plate 10, the second phase plate 11, and the third phase plate 12 , the first reflector 13, the second reflector 14, the second dichroic prism 15, the first dichroic mirror 16, the second dichroic mirror 17, the microscope objective lens 18, the nano translation stage 19, the bandpass filter 20 , field lens 21, detector 22.

[0036] The projection device includes: a first dichroic mirror 16 , a second dichroic mirror 17 and a microscope objective lens 18 . The detection imaging system is composed of a first dichroic mirror 16 , a second dichroic mirror 17 , a m...

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Abstract

The invention discloses a stimulated emission depletion (STED) microscopy method based on tangential polarized light. The stimulated emission depletion microscopy method comprises the following steps of: respectively converting a first laser beam and a second laser beam into first tangential polarized light and second tangential polarized light; projecting the first tangential polarized light onto a fluorescence sample to form an exciting light spot after phase modulation; decomposing the second tangential polarized light into a first working beam and a second working beam, and carrying out phase modulation on the first working beam and the second working beam to form a first STED beam and a second STED beam; projecting the first STED beam and the second STED beam onto the fluorescence sample to form a corresponding STED light spot through incoherence addition, wherein the central point of the STED light spot coincide with the central point of the exciting light spot to obtain a focusing light spot; and collecting fluorescence emitted by the fluorescence sample under the action of the focusing light spot, and obtaining a microscopic image after processing. The invention also discloses an STED microscopy device based on the STED microscopy method. According to the STED microscopy method and the STED microscopy device, a higher resolution can be realized under the same working light intensity.

Description

technical field [0001] The invention relates to the field of super-resolution microscopy, in particular to a stimulated emission loss microscopy method and device based on tangentially polarized light. Background technique [0002] According to Abbe's diffraction limit theory, the size of the spot formed by the beam after being focused by a general lens can be expressed as Where λ is the wavelength of the incident beam, and NA is the numerical aperture of the lens used, so the limiting resolution of conventional far-field fluorescence microscopes is generally limited to about half a wavelength. [0003] In order to break through the limitation of Abbe's diffraction limit and enable fluorescence microscopy to achieve super-resolution microscopy in the far field, researchers have proposed a variety of super-resolution microscopy methods. Among them, stimulated emission depletion microscopy (STED: Stimulated Emission Depletion Microscopy) is currently the most mature and wide...

Claims

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Application Information

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IPC IPC(8): G01N21/64G02B21/36G02B27/28
Inventor 匡翠方李帅薛懿顾兆泰刘旭
Owner ZHEJIANG UNIV
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