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Treatment liquid composition, treatment liquid kit and method for simultaneous biological organ transparentizing and immunolabeling

A liquid treatment and transparent technology, which is applied in biological testing, preparation of samples for testing, material analysis by optical means, etc. Rapid, immunofluorescence effect determination

Active Publication Date: 2020-02-07
HEFEI INSTITUTES OF PHYSICAL SCIENCE - CHINESE ACAD OF SCI
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Problems solved by technology

Both Clarity and Cubic remove the cell membrane and allow macromolecules to enter, but the process is cumbersome and takes nearly 1-2 weeks; iDisco uses organic solvents, which have poor fluorescence compatibility, make the tissue hard and brittle, and have a pungent odor that is harmful to the human body. Harmful; ACT-presto uses pressure, requires special equipment, and requires a complicated pretreatment process before immunolabeling

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  • Treatment liquid composition, treatment liquid kit and method for simultaneous biological organ transparentizing and immunolabeling
  • Treatment liquid composition, treatment liquid kit and method for simultaneous biological organ transparentizing and immunolabeling
  • Treatment liquid composition, treatment liquid kit and method for simultaneous biological organ transparentizing and immunolabeling

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Embodiment Construction

[0044] The invention provides a treatment liquid for making intact organs transparent, which contains carbohydrates, urea, and nonionic surfactants. The sugar is selected as a monosaccharide with a relatively high refractive index such as fructose (RI is 1.48), which can achieve the matching of the internal refractive index of the biological sample and make the tissue more transparent. However, the highest RI cannot exceed 1.48 for the mixture of sugars as a solution, which is not enough to achieve the matching of the refractive index of the components inside the tissue. Therefore, it is necessary to fully hydrate some solid components in the tissue, or remove some tissue components, such as cell membrane lipids. Urea breaks the interaction of non-covalent bonds and causes reversible denaturation of proteins. Its addition increases the solubility of membrane proteins in the sample, fully hydrates them, and reduces their refractive index. However, using only sugar and urea, it...

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Abstract

The invention relates to a treatment liquid composition, a treatment liquid kit and a method for simultaneous biological organ transparentizing and immunolabeling. The treatment liquid composition according to the present invention comprises a transparent treatment liquid and a fluorescently labeled antibody in a volume ratio of 1: 10 to 1: 500 to the transparent treatment liquid, wherein the transparent treatment liquid comprises a saccharide, urea and a nonionic surfactant. Biological macromolecules such as antibodies are allowed to enter organs while transparentizing the whole organs (brain, spinal cord, articular cartilage, (tumor-bearing) organs and the like), and direct or indirect immunolabeling on the whole organ scale is achieved.

Description

technical field [0001] The present invention relates to a method for clearing and immunolabeling whole organs of biological tissues, in particular to a treatment solution and method for direct or indirect immunofluorescent labeling of whole organs such as brain, spinal cord, articular cartilage, and viscera while being cleared. Background technique [0002] The tissue transparent method can realize large-scale high-resolution detection and three-dimensional structural reconstruction of tissue morphology without tissue damage. Compared with traditional tissue sectioning and staining methods, it has a significant improvement. Of all the tissue clearing methods currently developed, only some tissue clearing protocols allow whole-organ immunolabeling, such as iDisco, Clarity, Cubic, ACT-presto, etc. Both Clarity and Cubic remove the cell membrane and allow macromolecules to enter, but the process is cumbersome and takes nearly 1-2 weeks; iDisco uses organic solvents, which have...

Claims

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Application Information

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IPC IPC(8): G01N21/64G01N33/533
CPCG01N21/6428G01N21/6486G01N33/533G01N21/64G01N1/30G01N33/53G01N1/28
Inventor 钟凯徐金勇汪增荣
Owner HEFEI INSTITUTES OF PHYSICAL SCIENCE - CHINESE ACAD OF SCI
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