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Double beam plate lighting microscan imaging method and microscope

A scanning imaging, dual-beam technology, used in microscopy, fluorescence/phosphorescence, optics, etc., can solve the problems of limited imaging rate, inability to use beams, and limited exposure mode of imaging devices, to reduce weight, achieve imaging speed, and improve Imaging effect

Active Publication Date: 2014-11-19
HUAZHONG UNIV OF SCI & TECH
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Problems solved by technology

[0003] The traditional confocal light-sheet microscopic imaging technology is limited by the exposure mode of the imaging device. This scanning method cannot achieve full frame rate readout and cannot use the beam scanned in the return trip, which greatly limits the imaging rate. Limits overall system imaging rate

Method used

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  • Double beam plate lighting microscan imaging method and microscope
  • Double beam plate lighting microscan imaging method and microscope
  • Double beam plate lighting microscan imaging method and microscope

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Embodiment Construction

[0033] The invention provides a micro-scanning imaging method of double-beam light sheet illumination, comprising the following steps:

[0034] The scanning laser output by the same light source is divided into the scanning laser in the first direction and the scanning laser in the second direction;

[0035] The scanning laser light in the first direction and the scanning laser light in the second direction are vertically projected on the sample to excite the fluorescence of the sample; and the projection direction of the scanning laser light in the first direction is opposite to that in the second direction;

[0036] Detect and collect the fluorescence excited on the sample and expose it for imaging;

[0037] Move the sample and repeat the above process until the scan acquires a complete image of the sample;

[0038] Wherein, the scanning laser in the first direction and the scanning laser in the second direction scan synchronously, and the scanning speed is the same;

[00...

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Abstract

The invention belongs to technical field of microscan, and discloses a double beam light sheet lighting microscan imaging method. The method comprises the steps: dividing scanning laser output by an identical light source into first directional scanning laser and second directional scanning laser; ensuring that the two paths of scanning laser are vertically projected onto a sample to motivate sample fluorescent light, and the projection directions are opposite; detecting and collecting fluorescent light motivated by the sample and performing exposure imaging; shifting the sample, and repeating the process till a complete sample image is obtained through scanning, wherein the two paths of scanning laser are synchronous, a light sheet laser sample is produced, and the projection direction is in the x-axis direction; the scanning direction is in the y-axis direction; the fluorescent light collected direction of the sample is in the z-axis direction; scanning initial positions of the two paths of scanning laser are in the center line of a scanning array plane; scanning directions of the two paths of scanning laser are opposite. The method performs light sheet lighting imaging on the transparent sample through double beam scanning laser, so as to obtain higher imaging speed, and enhances the imaging quality through image frequency domain weighted superposition.

Description

technical field [0001] The invention relates to the technical field of micro-imaging, in particular to a micro-scanning imaging method and a microscope for double-beam light sheet illumination. Background technique [0002] The light sheet illumination microscope uses a micron-scale excitation light sheet to excite the fluorescence of the sample, and realizes the fast fluorescence signal detection of the sample by means of full-field detection. Light sheet illumination microscopic imaging uses linear illumination instead of traditional point scanning imaging, so it has higher imaging rate, higher signal-to-noise ratio and lower photobleaching and phototoxicity effects, suitable for hours or even several days In vivo bioimaging. [0003] The traditional confocal light-sheet microscopic imaging technology is limited by the exposure mode of the imaging device. This scanning method cannot achieve full frame rate readout and cannot use the beam scanned in the return trip, which ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64G02B21/36
Inventor 付玲杨哲梅丽龚辉骆清铭
Owner HUAZHONG UNIV OF SCI & TECH
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