Method for detecting form of neuron dendritic spine in brain slice

A detection method and neuron technology, applied in the field of brain neuron research, can solve the problems of complex steps, long process, high cost, etc., and achieve the effect of simple operation steps, stable dyeing effect, and avoiding high background interference

Pending Publication Date: 2021-10-01
WEST CHINA HOSPITAL SICHUAN UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0006] The technical problem to be solved by the present invention is: the existing method for staining dendritic spines in the brain has complex steps, long process, high cost, and the need to use highly toxic reagents and special instruments that are harmful to the human body.

Method used

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  • Method for detecting form of neuron dendritic spine in brain slice
  • Method for detecting form of neuron dendritic spine in brain slice

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Embodiment 1

[0038] 1. Brain Slice Preparation

[0039] 1.1 Prepare a 1.5% paraformaldehyde-PBS solution: Dissolve 15 g of paraformaldehyde in 1 L of PBS solution (0.01 mol / L phosphate buffer, pH=7.4, the PBS mentioned below are all the same solution).

[0040] 1.2 After the mice were deeply anesthetized, the heart was perfused with PBS solution for 5 minutes, and then fixed with 1.5% paraformaldehyde-PBS solution for 20 minutes.

[0041] 1.3 Take out the complete brain from the cranial cavity, and immediately place it on a vibrating microtome to slice with a thickness of 300 μm. After slicing, take a clean six-hole plate and soak the brain slices in PBS solution for later use.

[0042] 2. DIL staining

[0043] 2.1 Prepare DIL stock solution: fully dilute the DIL solid with DMSO to a stock solution with a concentration of 20mmol / L, and store the stock solution at -20°C.

[0044] 2.2 Prepare DIL working solution: dilute 20mmol / L DIL stock solution in 1.5ml EP tube with PBS solution to 5μ...

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Abstract

The invention belongs to the field of brain neuron research, and particularly relates to a method for detecting the form of a neuron dendritic spine in a brain slice. In order to solve the technical problems that the existing method for dyeing dendritic spines in the brain is complex in steps, long in process and high in cost, and highly toxic reagents and special instruments harmful to a human body need to be used, the invention provides a rapid detection method for neuron dendritic spines in a brain slice. According to the invention, the low-concentration dye and the high-strength vortex are adopted, so that the DIL working solution forms extremely small particles, and high background interference caused by DIL solid dye residues is avoided by controlling the contact time of the DIL working solution and the brain slice and an appropriate observation time window, so that a single neuron cell is clearly distinguished. According to the method, the original DIL scattering method process is greatly simplified, special instruments are not needed, the operation steps are simple, the dyeing effect is stable, and the method is suitable for laboratory popularization.

Description

technical field [0001] The invention belongs to the field of brain neuron research, and in particular relates to a method for detecting the morphology of neuron dendritic spines in brain slices. Background technique [0002] Dendritic spine (Dendrite spine) is located on the dendrites of neurons to form synapses, which is closely related to learning and memory. Dendritic spines are crucial in the connections between neurons in the brain. As the disease progresses, dendritic spines undergo both quantitative and morphological changes, which can have a significant impact on brain function. Explaining the altered plasticity of dendritic spines under pathological conditions will help us understand the learning and memory processes of the normal brain and the pathogenesis of various neurologically related diseases. [0003] Visualizing the shape of dendritic spines of nerve cells through various methods for subsequent analysis of the shape and density of dendritic spines is curr...

Claims

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Application Information

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IPC IPC(8): G01N1/30G01N1/34G01N1/28
CPCG01N1/30G01N1/34G01N1/286G01N2001/302G01N2001/305G01N2001/2873
Inventor 吴佳慧陈婵
Owner WEST CHINA HOSPITAL SICHUAN UNIV
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