Method for rendering tissue transparent, reagent for rendering tissue transparent, and tissue observation method
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Publication Number
US-12474239-B2
Publication Date
2025-11-18
Expiration Date
Abstract
A method for rendering tissue transparent, comprising a procedure of immersing the tissue in a water-soluble solvent comprising 2,2′-thiodiethanol and at least one of glycerol and a nonionic organoiodine compound is provided as a technique capable of sufficiently rendering various organs transparent without causing their changes by a simple operation without using any poisonous or dangerous organic solvent. In the method for rendering tissue transparent, a mixed solvent of 2,2′-thiodiethanol, glycerol, and a nonionic organoiodine compound aqueous solution is preferably used as the water-soluble solvent.
Core Innovation
The invention provides a method for rendering tissue transparent. The method comprises fixing a tissue and immersing the fixed tissue in a water-soluble solvent comprising 2,2′-thiodiethanol, a nonionic organoiodine compound, and optionally glycerol.
The tissue comprises a tissue surface and a tissue site, and the tissue site is located at a depth of 1 mm or more below the tissue surface. The observing step comprises detecting fluorescence emitted from the tissue site by using a fluorescence microscope, a fluorescence stereomicroscope, a confocal microscope, or a multiphoton microscope.
The water-soluble solvent and observing workflow are configured to transparentize fixed tissues while enabling fluorescence observation of deeper tissue sites. The document describes that fluorescence from fluorescent proteins is preserved during transparentization.
The document describes that the approach is applied to diverse organs and tissues, and includes fluorescence labeling before or after transparentization and detection by confocal or multiphoton microscopy. The transparentization is contrasted with conventional approaches involving harmful organic solvents, and the document reports comparisons related to transparency, swelling or weakening, imaging depth, and fluorescence retention.
Claims Coverage
The independent claim contains three inventive aspects: solvent formulation for transparentization, fixing followed by immersion in the solvent, and fluorescence-based observation with a depth condition of 1 mm or more below the tissue surface.
Water-soluble transparentization solvent with 2,2′-thiodiethanol and nonionic organoiodine
A method for rendering tissue transparent that immerses the fixed tissue in a water-soluble solvent comprising 2,2′-thiodiethanol, a nonionic organoiodine compound, and optionally glycerol.
Fluorescence observation of a deeper tissue site
A method in which observing comprises detecting fluorescence emitted from a tissue site located at a depth of 1 mm or more below the tissue surface using a fluorescence microscope, a fluorescence stereomicroscope, a confocal microscope, or a multiphoton microscope.
Fixing followed by immersion in the solvent
A method that includes a step of fixing a tissue and then immersing the fixed tissue in the water-soluble solvent comprising 2,2′-thiodiethanol and a nonionic organoiodine compound.
Overall, claim coverage centers on fixing tissue, transparentization via a water-soluble solvent containing 2,2′-thiodiethanol and a nonionic organoiodine compound, and fluorescence detection from a tissue site located 1 mm or more below the tissue surface using fluorescence-based microscopes including confocal or multiphoton microscopy.
Stated Advantages
Fluorescence from fluorescent proteins is preserved during transparentization.
The approach enables fluorescence observation of deeper tissue sites.
The document reports comparisons related to transparency, swelling or weakening, imaging depth, and fluorescence retention.
The transparentization is contrasted with conventional approaches involving harmful organic solvents.
Documented Applications
The approach is applied to diverse organs and tissues.
Fluorescence labeling before or after transparentization.
Detection by confocal microscopy or multiphoton microscopy.
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