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Super-resolution fluorescence microscopic method and apparatus based on photoactivation and structured light illumination

A technology of super-resolution fluorescence and structured light illumination, applied in the field of super-resolution fluorescence microscopy methods and devices, can solve the problems of photobleaching, difficult to apply in the field of super-resolution imaging, etc., and achieve the effect of high lateral resolution

Active Publication Date: 2016-11-16
ZHEJIANG UNIV
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  • Application Information

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Problems solved by technology

Although SSIM can increase the lateral resolution to less than 100nm by increasing the illumination light power, it inevitably brings about the problem of photobleaching, making it difficult to apply to the field of super-resolution imaging of live cells.

Method used

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  • Super-resolution fluorescence microscopic method and apparatus based on photoactivation and structured light illumination
  • Super-resolution fluorescence microscopic method and apparatus based on photoactivation and structured light illumination
  • Super-resolution fluorescence microscopic method and apparatus based on photoactivation and structured light illumination

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Embodiment Construction

[0062] like figure 1 Shown: A super-resolution fluorescence microscopy device based on photoactivation and structured light illumination, including:

[0063] A light source module 1, and a single-mode optical fiber 2, a collimator lens 3, a first plane reflector 4, and a second plane reflector 5 arranged in sequence in the optical path of the light source module 1;

[0064] The modulation unit includes a first half wave plate 6 and a first polarization beam splitting cube 7; a second half wave plate 8 and a second polarization beam splitting cube 11 located on the transmission optical path of the first polarization beam splitting cube 7 , the first quarter-wave plate 12, the first mirror 15, the first piezoelectric ceramic 25, the second mirror 16, the third mirror 18 and the fourth mirror 20; located in the first polarization beam splitting cube 7 The third half-wave plate 9, the third polarization beam splitting cube 10, the second quarter-wave plate 13, the fifth mirror 14...

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Abstract

The invention discloses a super-resolution fluorescence microscopic apparatus based on photoactivation and structured light illumination. The apparatus comprises a light source module having a first laser for fluorescence activation, a second laser for fluorescence excitation, and a frequency selection switching module for switching between the two lasers; a regulation unit for regulating a light beam output from the light source module to two beams of p polarized light and two beams of s polarized light capable of interference and for changing the interference phase difference of the two groups of light beams; a dichroic mirror, on the surface of which interference fringes are formed via the two beams of p polarized light and two beams of s polarized light, and which reflects a mesh structured light illumination comprising bright spots and dark spots in array distribution and taken as an irradiation sample; and an imaging unit comprising a converging module for changing the spacing between interference fringes, a microobjective projecting a light beam emerging from the converging module to the microobjective of a sample, and a camera for conducting stimulated radiation fluorescence imaging of the sample. The invention further discloses a super-resolution fluorescence microscopic method based on photoactivation and structured light illumination.

Description

technical field [0001] The invention relates to the field of optical super-resolution microscopy, in particular to a super-resolution fluorescence microscopy method and device based on photoactivation and structured light illumination. Background technique [0002] As a technology with nanoscale imaging capabilities, super-resolution fluorescence microscopy plays an irreplaceable role in the field of protein dynamics in living cells, and is an important means to reveal the phenomena and laws of basic life activities. However, the current super-resolution technology is limited by various principles and technical factors, and it is still insufficient in super-resolution imaging of living cells. [0003] At present, the relatively successful super-resolution fluorescence microscopy techniques mainly include the following: single-molecule fluorescence imaging (PALM and STORM), stimulated radiation depletion microscopy (STED), and structured illumination microscopy (SIM and SSIM)...

Claims

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Application Information

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IPC IPC(8): G01N21/64
CPCG01N21/6402G01N21/6458
Inventor 刘旭陈友华匡翠方朱大钊
Owner ZHEJIANG UNIV
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