Light-activated positioning microscopic imaging system based on optical fiber evanescent field illuminator
A technology of illuminator and evanescent field, which is applied in the field of light-activated positioning microscopic imaging system, can solve the problems of inability to observe the upper surface and side of cells, increase costs, etc., and achieve the effects of convenient adjustment, lower system cost, and simplified system optical path
Inactive Publication Date: 2011-01-19
HUAZHONG UNIV OF SCI & TECH
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[0006] However, the evanescent field only exists in the range of about 100nm from the adherent surface of the sample, so light-activated localization microscopy can generally only be used to observe the shallow information of the adherent surface of the cell, but cannot observe the upper surface of the cell that may have biological functions and side; at the same time, the high numerical aperture objective lens used in order to obtain better evanescent field increases the cost of the system
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Embodiment 1
[0018] Embodiment 1: The super-resolution microscopic imaging of biological cells is realized by using a light-activated localization microscopic imaging system based on a fiber optic evanescent field illuminator. figure 1 It is a schematic diagram of the system structure of the light-activated localization microscopy imaging system based on the optical fiber evanescent field illuminator. The experimental cell is the BK channel of hek293 cells labeled with mEosFP photoconversion fluorescent protein. The active laser uses a laser with a wavelength of 405nm, and the imaging laser uses a laser with a wavelength of 561nm. The optical axes of the activation laser and the imaging laser are adjusted to coincide with each other by means of the mirror 6 and the dichromatic mirror 5 . The active and imaging lasers are coupled into the fiber optic evanescent field illuminator by using lens 4 . The manipulator 3 can adjust the three-dimensional position of the optical fiber evanescent f...
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Abstract
The invention relates to a light-activated positioning microscopic imaging system based on an optical fiber evanescent field illuminator. Activation lasers emitted by an activation laser sequentially pass through a neutral filter, a shutter and a controller thereof, a reflective mirror, a dichroscope and a lens and then enter the optical fiber evanescent field illuminator; imaging lasers emitted by an imaging laser sequentially pass through the neutral filter, the shutter and the controller thereof, the dichroscope and the lens and then enter the optical fiber evanescent field illuminator; the optical fiber evanescent field illuminator is fixed on a controlling device and is arranged in a sample pool of an inverted fluorescence microscope, and a detector EMCCD (Electron Multiplying Charge-Coupled Device) collects fluorescence signals collected by an objective of the inverted fluorescence microscope. According to the invention, the illuminating light path is separated from the imaging light path so as to simplify the light path, the illumination is carried out by using the optical fiber evanescent field illuminator without an objective with high numerical aperture so as to reduce the system cost; meanwhile each surface of a cell sample is imaged by controlling the position, the depth and the angle of the optical fiber evanescent field illuminator.
Description
technical field [0001] The invention relates to a photoactivation positioning microscopic imaging system based on an optical fiber evanescent field illuminator, in particular to a photoactivation positioning microscopic imaging system capable of performing multi-directional illumination imaging on a sample. Background technique [0002] In the field of biological sciences, precise positioning of specific proteins plays an important role in studying the location and function of proteins. Electron microscopy and non-optical scanning probe microscopy can achieve nanoscale resolution, but they cannot specifically label and study individual protein molecules, nor can they be used to observe and study living organisms. [0003] Optical microscopy imaging can be used for non-destructive in vivo detection and single-molecule detection, and can provide important optical information such as polarization state and spectrum. These advantages make optical microscopy imaging widely used i...
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IPC IPC(8): G01N21/64
CPCG01N21/648G01N21/6458
Inventor 吕晓华曾绍群骆清铭黄振立付玲
Owner HUAZHONG UNIV OF SCI & TECH