Optical transparency method for quickly acquiring three-dimensional data of large-volume sample

Abstract

The invention discloses a method for rapidly acquiring three-dimensional data of a large-volume biological sample. The method comprises the following steps: S1, a first immersion step; s2, an embedding step; s3, a second immersion step; s4, a sample installation step; s5, an optical imaging step; and s6, a slicing step; wherein the S5 and S6 are circularly and alternately carried out until a three-dimensional image of the biological sample tissue is obtained; the light transparent solution comprises the following raw materials in percentage by mass: 20-40% of sugar or alcohol or sugar-alcohol compound, 10 to 30% of urea, 0-30% of a penetration enhancer and the balance of water. According to the invention, the optical transparent solution and the microsection tomography technology are combined for use; optical imaging and microsection are alternately carried out, and meanwhile, the optical transparent solution is used for replacing liquid between tissues of a biological sample in real time; therefore, the refractive index of the biological sample tissue is quickly balanced, the depth of single imaging is expanded, and the slicing frequency of the whole sample is remarkably reduced. Therefore, the applicable sample range of light transparency and the continuous acquisition efficiency of three-dimensional data are greatly improved.

Description

technical field [0001] The invention relates to the technical field of biomedical optical imaging, in particular to an optically transparent method for rapidly acquiring three-dimensional data of a large-volume sample. Background technique [0002] The brain is one of the most important and complex organs in humans. Its structure is complex and neurons are abundant. But at present, we know very little about the function and structure of the brain. Brain science research will help scientists to better decipher brain function and brain diseases, and brain research can promote the development of artificial intelligence. Whole-brain three-dimensional imaging technology is often used in the study of neural circuits. A neural circuit is an intricate series of circuits in which neurons are interconnected by synaptic structures. These circuits perform the higher functions of the brain, controlling behavior and emotion, and the study of neural circuits is considered fundamental t...

AI Technical Summary

Problems solved by technology

Organic reagents have high transparency efficiency, but have the disadvantages of large sample deformation and poor fluorescence retention. Hydrophilic reagents have the characteristics of low deformation and high fluorescence retention, but their transparency efficiency is low, which is difficult for large-volume samples such as macaque brain. are completely transparent and are therefore only commonly used for 3D imaging of small volume samples

For example, the imaging device and method disclosed in Chinese patent 201610315684.3 to quickly obtain three-dimensional structure information and molecular phenotype information of large samples mainly uses vibration sectioning to remove the imaged surface and wide-field microscopy to obtain surface data of newly exposed samples. However, due to the Opaque and uneven refractive index lead to shallow imaging depth. In order to ensure cutting quality, the cutting thickness is often greater than the imaging depth. For example, each cutting is 50 microns, and the imaging surface is 10 microns. reconstruction

Images