High-speed WFOV (wide field of view) CARS (coherent anti-stokes raman scattering) microscope system and method

A microscopic system and wide field of view technology, applied in microscopes, optics, instruments, etc., can solve the problems of slow imaging speed and photodamage of living biological tissues, and achieve the effect of small photodamage, fast speed and low energy density of samples

Active Publication Date: 2013-03-27
BEIJING LUSTER LIGHTTECH
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Problems solved by technology

[0014] In order to solve the technical problems of the existing CARS microscopy technology, such as slow imaging speed and serious photodamage to living biological tissues, the present invention provides a high-speed wide-field coherent anti-Stokes Raman scattering microscopy system and method

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  • High-speed WFOV (wide field of view) CARS (coherent anti-stokes raman scattering) microscope system and method
  • High-speed WFOV (wide field of view) CARS (coherent anti-stokes raman scattering) microscope system and method
  • High-speed WFOV (wide field of view) CARS (coherent anti-stokes raman scattering) microscope system and method

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Embodiment 1

[0062] Figure 4 It is the CARS imaging experiment of the device of the present invention on C. elegans nematode eggs. In the experiment, the focusing objective lens is a 10X microscopic objective lens, NA=0.4, the detection objective lens is a 40X microscopic objective lens, NA=0.85, and the scale is 10 μm. The pump light laser is a Ti:Sapphire laser with a pulse width of 3 ps, a repetition rate of 76 MHz, and a wavelength of 700- 980nm adjustable. The OPO laser is used as the Stokes laser, the pulse width is 6ps, the repetition frequency is 76MHz, and the wavelength is adjustable from 680-990nm. During the experiment, the wavelength difference of the two lasers was adjusted to match the CH of the fat molecule. 2 Bond resonance, resonance frequency 2850cm -1 . The cell membrane of the C. elegans nematode egg and the adipose tissue inside can be clearly observed. The image contrast is high, and the EMCCD exposure time is 0.5 seconds.

Embodiment 2

[0064] Figure 5It is the experimental result of moving a polystyrene ball with a diameter of 6 μm in water and performing CARS imaging. In the experiment, the focusing objective lens is a 10X microscopic objective lens, NA=0.4, the detection objective lens is a 40X microscopic objective lens, NA=0.85, and the scale is 10 μm. The pump light laser is a Ti:Sapphire laser with a pulse width of 3 ps, a repetition rate of 76 MHz, and a wavelength of 700- 980nm adjustable. The OPO laser is used as the Stokes laser, the pulse width is 6ps, the repetition frequency is 76MHz, and the wavelength is adjustable from 680-990nm. During the experiment, the wavelength difference between the two lasers was adjusted to make it resonate with the CH bond of the polystyrene molecule, and the resonance frequency was 3050cm -1 . The dynamic process of moving polystyrene pellets in water can be clearly observed. The image acquisition rate of 33fps can be realized. The present invention is fully ...

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Abstract

The invention relates to a high-speed WFOV (wide field of view) CARS (coherent anti-stokes raman scattering) microscope system and a high-speed WFOV CARS microscope method. In the invention, pumping laser and Stokes light laser which are totally coincident in the aspects of space and time are subjected to weak convergence, so that a sample generates a CARS signal; and the CARS signal enters a CCD (Charge Coupled Device) camera through an optical filter and a cylindrical lens so as to obtain a clear CARS image. The invention utilizes the CARS signal to image and relates to an imaging technology based on the vibration characteristic of energy level inside molecules. The high-speed WFOV CARS microscope system and the high-speed WFOV CARS microscope method can be used for detecting chemical compositions of the sample and can be used for carrying out imaging on a single cell, even a single organelle. The requirements of most of biological experiments are totally met. The technical problems of low imaging speed, series photic damage to living biological tissues and the like of the existing CARS microtechnique are solved. Compared with the common fluorescence microscopy, the high-speed WFOV CARS microscope system and the high-speed WFOV CARS microscope method have the advantages that an external fluorescent probe does not need to be used, and influence on the molecular structure of the sample cannot be caused.

Description

technical field [0001] The invention relates to a high-speed and wide-field coherent anti-Stokes Raman scattering (CARS) microscopic system, which can be widely used in research in the fields of biology, medicine, biophysics, material chemistry and the like. Background technique [0002] In the research of materials chemistry and biology, scientists often need microscopes with high spatial resolution and the ability to detect the molecular composition of materials or biological cells. With the emergence of various fluorescent probes, different fluorescent probes can be used to mark different positions inside the sample, so that the molecular composition of the sample can be detected. Fluorescence microscopy has become a powerful tool for imaging chemical and biological samples. However, there are two disadvantages in fluorescence microscopy: the photobleaching effect of fluorescent probes and the influence of fluorescent probes on the molecular structure of samples. [000...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G02B21/36G02F1/35G02B27/10
Inventor 雷铭姚保利严绍辉但旦叶彤
Owner BEIJING LUSTER LIGHTTECH
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