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US 10,309,879 B2Granted
Expansion microscopy
AssigneeMassachusetts Institute of Technology
InventorsFei Chen, Paul Warren Tillberg, Edward Stuart Boyden
Life of this patent
Estimating the term…
Abstract
The present invention relates to an enlarged sample of interest for microscopy and methods for enlarging a sample of interest and the optical imaging of a sample of interest with resolution better than the classical microscopy diffraction limit, by synthesizing a swellable polymer network within a sample, it can be physically expanded, resulting in physical magnification.
Claims46
A method for enlarging a cell or solid tissue-sample, the method comprising:
- (a) labeling more than one biomolecule components of the sample with a trifunctional label, wherein the tri-functional label comprises an anchor capable of participating in polymerization and a visual label, further wherein the anchor is methacryloyl;
- (b) permeating the sample with a composition comprising sodium acrylate, acrylamide and N,N-methylenebisacrylamide;
- (c) polymerizing the composition within the sample to form a swellable material, wherein said polymerizing results in covalent crosslinking of the anchor to the swellable material to form a sample-swellable material complex;
- and
- (d) adding an aqueous solvent or liquid to cause the sample-swellable material complex to swell, thereby physically expanding the complex that results in an enlarged cell or tissue sample.
2 more elements
A method for preparing an enlargeable cell or solid tissue sample, the method comprising:
- (a) labeling more than one biomolecule components of the sample with a trifunctional label, wherein the tri-functional label comprises an anchor capable of participating in polymerization and a visual label, further wherein the anchor is methacryloyl;
- (b) permeating the sample with a composition comprising sodium acrylate, acrylamide and N,N-methylenebisacrylamide;
- (c) polymerizing the composition within the sample to form a polyelectrolyte hydrogel, wherein said polymerizing results in covalent crosslinking of the anchor to the polyelectrolyte hydrogel to form a cell or a tissue sample-polyelectrolyte hydrogel complex, thereby resulting in a sample prepared to be enlarged.
A method for isotropically enlarging a cell or solid tissue sample, the method comprising:
- (a) labeling more than one biomolecule components of the sample with a label, wherein the label comprises a biomolecule binding moiety and a polymerizable ethylenically unsaturated moiety;
- (b) permeating the sample with a composition comprising sodium acrylate, acrylamide and N,N-methylenebisacrylamide;
- (c) polymerizing the composition within the sample to form a swellable material, wherein said polymerizing results in covalent crosslinking of the polymerizable moiety to the swellable material to form a sample-swellable material complex;
- and
- (d) adding an aqueous solvent or liquid to cause the sample-swellable material complex to swell, thereby physically expanding the complex that results in an isotropically enlarged tissue sample.
2 more elements
A method for isotropically enlarging a cell or solid tissue sample, the method comprising:
- (a) labeling more than one biomolecule components of the sample with a trifunctional label, wherein the tri-functional label comprises an anchor capable of participating in polymerization and a visual label, further wherein the anchor is methacryloyl;
- (b) permeating the sample with a composition comprising precursors of a crosslinked hydrogel comprising at least one polyelectrolyte monomer and a covalent crosslinker;
- (c) polymerizing the composition within the sample to form a swellable material, wherein said polymerizing results in covalent crosslinking of the anchor to the swellable material to form a sample-swellable material complex;
- and
- (d) adding an aqueous solvent or liquid to cause the sample-swellable material complex to swell, thereby physically expanding the complex that results in an isotropically enlarged tissue sample.
2 more elements
A method for isotropically enlarging a cell or solid tissue sample, the method comprising:
- (a) contacting the sample with a tag comprising a binding moiety and a polymerizable moiety, wherein the tag binds a biomolecule component of the sample;
- (b) permeating the sample with a composition comprising precursors of a crosslinked hydrogel comprising at least one polyelectrolyte monomer and a covalent crosslinker;
- (c) polymerizing the composition within the sample to form a swellable material, wherein said polymerizing results in covalent crosslinking of the tag to the swellable material to form a sample-swellable material complex;
- and
- (d) adding an aqueous solvent or liquid to cause the sample-swellable material complex to swell, thereby physically expanding the complex that results in an isotropically enlarged tissue sample.
2 more elements
A method for isotropically enlarging a cell or solid tissue sample, the method comprising:
- (a) contacting the sample with a tag comprising a binding moiety and a polymerizable moiety, wherein the tag binds a biomolecule component of the sample;
- (b) permeating the sample with a composition comprising precursors of a polyelectrolyte hydrogel comprising at least one polyelectrolyte monomer and a covalent crosslinker;
- (c) polymerizing the composition within the sample to form a polyelectrolyte hydrogel, wherein said polymerizing results in covalent crosslinking of the tag to the swellable material to form a sample- polyelectrolyte hydrogel complex;
- and
- (d) adding an aqueous solvent or liquid to cause the sample-polyelectrolyte hydrogel complex to swell, thereby physically expanding the complex that results in an isotropically enlarged tissue sample.
2 more elements
A method for isotropically enlarging a cell or solid tissue sample, the method comprising:
- (a) contacting the sample with a tag comprising a binding moiety and an anchor, wherein the binding moiety binds to biomolecule components of the sample;
- wherein the anchor comprises a polymerizable moiety;
- (b) permeating the sample with a composition comprising at least one polyelectrolyte monomer and a covalent crosslinker;
- (c) initiating polymerization to form a hydrogel comprising the at least one polyelectrolyte monomer, covalent crosslinker and anchor, wherein the hydrogel is covalently bound to the biomolecule components of the sample to form a sample- hydrogel complex;
- and
- (d) adding an aqueous solvent or liquid to cause the sample-hydrogel complex to swell, thereby physically expanding the complex that results in an isotropically enlarged tissue sample.
3 more elements
Dependencies are inferred from the claim text.