Methods for phenotyping of intact whole tissues
AssigneeCalifornia Institute of Technology
InventorsViviana Gradinaru, Bin Yang
Life of this patent
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Abstract
In various embodiments, the present application teaches methods and compositions for tissue clearing in which whole organs and bodies are rendered macromolecule-permeable and optically-transparent, thereby exposing their cellular structure with intact connectivity. In some embodiments, the present application teaches PACT, a protocol for passive tissue clearing and immunostaining of intact organs. In other embodiments, the present application teaches RIMS, a refractive index matching media for imaging thick tissue. In yet other embodiments, the application teaches PARS, a method for whole-body clearing and immunolabeling.
Claims69
A method for modifying the structural and/or optical characteristics of a tissue, comprising:
- applying a fixing solution comprising paraformaldehyde (PFA) to the tissue, thereby forming fixed tissue, and
- applying a hydrogel monomer solution comprising acrylamide and phosphate buffered saline (PBS) to the fixed tissue, thereby forming hydrogel treated tissue.
A method for modifying the structural and/or optical characteristics of tissue in situ, comprising:
- introducing a fixing solution comprising paraformaldehyde (PFA) into the circulatory system of a subject, thereby forming fixed tissue within the subject;
- and
- introducing a hydrogel monomer solution comprising acrylamide and phosphate buffered saline (PBS) into the circulatory system of the subject, thereby forming hydrogel-treated tissue within the subject.
A composition, comprising:
- (1) 1-N,3-N-bis(2,3-dihydroxypropyl)-5-[N-(2,3-dihydroxypropyl)acetamido]-2,4,6-triiodobenzene-1,3-dicarboxamide or 5-(N-2,3-Dihydroxypropylacetamido)-2,4,6-triiodo-N,N′-bis(2,3-dihydroxypropyl)isophthalamide, (2) phosphate buffer;
- (3) tween-20;
- (4) sodium azide;
- and optionally (5) sodium hydroxide.
A method for modifying the structural and/or optical characteristics of brain tissue in situ, comprising:
- introducing a fixing solution comprising paraformaldehyde (PFA) into the cerebrospinal fluid (CSF) of a subject, thereby forming fixed brain tissue within the subject;
- and
- introducing a hydrogel monomer solution comprising acrylamide and phosphate buffered saline (PBS) into the CSF of the subject, thereby forming hydrogel-treated brain tissue within the subject.
A method for modifying the structural and/or optical characteristics of spinal cord tissue in situ, comprising:
- introducing a fixing solution comprising paraformaldehyde (PFA) into the cerebrospinal fluid (CSF) of a subject, thereby forming fixed spinal cord tissue within the subject;
- and
- introducing a hydrogel monomer solution comprising acrylamide and phosphate buffered saline (PBS) into the CSF of the subject, thereby forming hydrogel-treated spinal cord tissue within the subject.
A method as described in the specification.
Dependencies are inferred from the claim text.