High-resolution light sheet microscopic imaging system for observing free-moving zebra fish

Abstract

The invention discloses a high-resolution light sheet microscopic imaging system for observing free-moving zebra fish. The system comprises a light source module, a transverse view field expansion module, a scanning module, an illumination module and a detection module which are sequentially arranged according to a light path, light generated by the light source module passes through the transverse view field expansion module to obtain an effective view field for expanding a final illumination area; the scanning module scans a linear light beam received from the transverse view field expansionmodule into a surface light beam; the illumination module and the detection module adopt a beam splitter, an illumination unit, a first illumination objective lens and a second illumination objectivelens to enable transmitted infrared light to carry structural information of a zebra fish sample to enter the detection module, and the detection module adopts an infrared camera and a complementarymetal oxide semiconductor camera to realize near-infrared light and fluorescence detection imaging respectively. The method has the advantages of no influence on the sample, high transverse and axialresolution and high imaging speed.

Description

technical field [0001] The technical field of optical microscopic imaging of the present invention particularly relates to a high-resolution light sheet microscopic imaging system for observing free-moving zebrafish. Background technique [0002] Zebrafish is a model organism with high homology to the human genome (favorable for establishing disease models), so it is a commonly used sample in biomedical research. Because its larvae are transparent throughout, they are suitable for imaging research by optical microscopy imaging systems. In the conventional in vivo imaging system for zebrafish, the zebrafish is fixed in agarose gel during observation to prevent its activities from affecting the imaging. The main reason is that in conventional imaging methods, resolution and imaging field of view are mutually restricted. When obtaining high-resolution images, the imaging field of view is often small, and the sample needs to be strictly fixed within the field of view. However,...

AI Technical Summary

Problems solved by technology

However, in order to ensure a large enough field of view, the numerical aperture of the objective lens used in this system is 0.04, resulting in low imaging resolution, which is not enough for high-resolution neuroimaging research

In addition, the detection light and illumination light of the above-mentioned imaging methods are coaxial, which brings problems that the axial resolution is generally low, and the phototoxicity and photobleaching of the sample are obvious, which is not conducive to long-term imaging of the sample.

However, the research on free-moving samples often requires long-term observation, so the above methods have certain disadvantages

Images