Skull photon transparentizing treatment method and application thereof

Abstract

The invention provides a skull photon transparentizing treatment method and application thereof. The treatment method comprises the following steps that a compact bone substance loosening agent is added to the skull surface for treatment, and the compact bone substance loosening agent is selected from collagenase, trypsin and disodium edetate dehydrate; the skull surface treated with the compact bone substance loosening agent is cleaned; a high-refraction-index water-soluble reagent is added to the cleaned skull surface for treatment, and the skull with the relatively uniform refraction index is obtained. The skull structure treated through the method becomes transparent, and can be used for living body two-photon microimaging, and high-resolution imaging of cortex nerves and microvascular structures is achieved.

Description

technical field [0001] The invention belongs to the technical field of biomedical optical imaging, and in particular relates to a skull optical transparency processing method and an application thereof. Background technique [0002] The long-term observation and manipulation of cortical cells is of great significance for the study of neurons, glial cells and microvasculature. With the development of transgenic technology and two-photon microscopic imaging, scientists can conduct research on the cortex at the level of living mammals. Repeated imaging over time to understand the complex environment of the nervous system. Especially to track the fate of individual synapses, and then study the regulation of sensory experience, learning and memory on the dynamic changes of dendritic spines. [0003] However, due to the high scattering properties of the skull above the cortex, the skull becomes very turbid, and it is difficult to focus photons in deep brain imaging. To solve this...

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Problems solved by technology

[0003] However, due to the high scattering properties of the skull above the cortex, the skull becomes very turbid, and it is difficult to focus photons in deep brain imaging. To solve this problem, researchers have mainly developed different cranial windows. The first is to use craniectomy During the operation, part of the skull was completely removed and the exposed meninges were covered with a cover glass. Due to the inevitable exposure of the brain, the physical and chemical properties of the cortex were changed, causing different degrees of inflammatory reactions, which affected the reliability and credibility of the experimental data. The second is to use skull thinning surgery to grind the skull to a thickness of less than 25μm, which is very difficult; the third is to combine the two operations to grind the skull to about 10-15μm, and then use a cap Covered with slides, but this type of cranial window is mainly used in the study of cortical microvasculature

It is precisely because of the shortcomings of these cranial window techniques, coupled with the complex surgical process, difficult surgery, and high requirements for experimenters, that severely limit the research of cortical neuroimaging.

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