DMD-based dual-mode optical super-resolution microscopic imaging device and method

A microscopic imaging and optical imaging system technology, which is applied in the field of dual-mode optical super-resolution microscopic imaging devices, can solve the problems of complex optical path system and cost reduction, and achieve cost reduction, lower threshold, high stability and ease of use Effect

Active Publication Date: 2017-09-15
SOUTH CHINA NORMAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, in their research, SMLM and SIM are still two independent parts. Multiple shutters or expensive acousto-optic tunable filters are required in the SMLM optical path to realize laser pulse modulation, and spatial light modulation elements are also required in the SIM optical path. The optical path system Very complex, cost not reduced

Method used

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  • DMD-based dual-mode optical super-resolution microscopic imaging device and method
  • DMD-based dual-mode optical super-resolution microscopic imaging device and method
  • DMD-based dual-mode optical super-resolution microscopic imaging device and method

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Embodiment 1

[0041] In this embodiment, the dual-mode optical super-resolution microscopic imaging device based on DMD is as follows: figure 1 As shown, among them, 1 is a semiconductor laser with a wavelength of 532nm, and 2 is a semiconductor laser with a wavelength of 405nm. Two kinds of lasers with different wavelengths can be selected according to actual imaging needs. 3, 4, 5, 6, 8, and 11 are plane mirrors with adjustable angles, 7 is a DMD chip, 9, 12, and 14 are lenses, 10 is a mask for diffraction spot gating, and 13 is an aperture stop , 15 is a 545nm long-pass dichroic mirror, 16 is an objective lens, 17 is a fluorescence filter, 18 is an imaging lens, and 19 is a sCMOS camera.

[0042] The super-resolution microscopic imaging device described in this embodiment is divided into the following parts according to the functions realized, which are two lasers, a laser modulation module, a microscope optical imaging system and an object stage, and the laser modulation module includes...

Embodiment 2

[0049] This embodiment is based on Figure 8 The shown device realizes single-molecule localization imaging SMLM, and its steps are:

[0050] In the same optical path system as in Embodiment 1, the single-hole mask shown in Figure 5 (b) is inserted at the position of the gate mask 10, and a single diffraction spot (such as Figure 8 shown). Control the DMD chip to switch between black and white stripes of two different spatial frequencies K1 and K2, so that the laser spots with wavelengths of 405nm and 532nm alternately pass through the small holes of the mask to obtain alternating light pulses (as shown in Figure 9 (a, b)). Use corresponding switchable fluorescent dyes, such as Alexa-fluo 532, to label cell samples. The dye molecules are randomly converted into active states under 405nm laser irradiation, and excited to emit fluorescence under 532nm laser irradiation, which is detected and recorded by a high-sensitivity camera down.

[0051] The DMD pattern controller cont...

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Abstract

The invention discloses a DMD-based dual-mode optical super-resolution microscopic imaging device and method. The device comprises multiple lasers of different wavelengths, a laser modulation module and a microscope optical imaging system. The laser modulation module comprises a DMD pattern generation module and a mask plate. The DMD pattern generation module is matched with the mask plate to gate different diffraction light spots so that the objective of realizing two types of super-resolution microscopic technology imaging of SIM and SMLM by sharing the same optical path can be achieved. The DMD has the response speed and the stability higher than those of a mechanical shutter and can be used for wavelength separation so that the cost is lower and use is easier in comparison with the acousto-optic tunable filter, the optical path system is enabled to be more compact and the cost of the system can be greatly reduced. Implementation of different imaging modes depends on multiplexing of the DMD. Using the DMD is put forward for the first time, and two technologies of SIM and SMLM are realized in the same optical path so that advantage complementation of the two super-resolution imaging technology of SIM and SMLM is facilitated, and the device and the method can be used for the research of multiple cell biological problems.

Description

technical field [0001] The invention belongs to the field of optical microscopy technology, and specifically relates to a DMD-based dual-mode optical super-resolution microscopic imaging device and method, which can realize the modulation of laser light sources required for two-mode imaging by using a digital micromirror device (DMD) . Background technique [0002] Optical microscopy allows the observation of samples grown in a natural environment without damage, but is limited by the diffraction limit. The lateral resolution is only half a wavelength, and it is impossible to obtain information on nanoscale intracellular structures. In order to improve the spatial resolution of the far-field optical microscope, people began to study super-resolution microscopy and made great progress in recent years. academic research. Various existing super-resolution imaging technologies have broken through the diffraction limit of optical imaging through different mechanisms: single mol...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G02B27/58G01N21/64
CPCG01N21/6458G01N2021/6478G02B27/58
Inventor 詹求强吴秋生马剑涛王保举李彦慧
Owner SOUTH CHINA NORMAL UNIVERSITY
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