Swellable photopolymerized hydrogels for expansion microscopy
AssigneeThe Regents of the University of Colorado, a body
InventorsKristi Anseth, Kemal Arda Gunay, Laura J. Macdougall
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Abstract
Compositions and methods for preparing and using a swellable polymer network for expansion microscopy (ExM) applications. The swellable polymer network can be prepared on demand using external stimuli for ExM applications. A variety of external stimuli can be employed, including light irradiation, pH, temperature, and magnetic fields. Network preparation using light irradiation is particularly advantageous, as it allows tuned control over the network properties through the variation of time and dose of light irradiation. In other words, light irradiation can be employed with a wavelength and intensity that enables a polymer network formation.
Claims22
1-85. (canceled)
A method for preparing a sample of interest for expansion microscopy comprising the steps of:
- i. fixing a sample of interest;
- ii. labelling a biological cue of interest within the sample of interest with one or more labelling groups;
- iii. functionalizing the sample of interest with a tethering group;
- iv. permeating the sample of interest with a swellable, photopolymerizable hydrogel solution;
- v. polymerizing the hydrogel solution via light irradiation, thereby allowing it to embed and tether the sample of interest into the hydrogel network;
- vi. digesting or removing the sample of interest with a solution using a digestion method that retains and preserves the spatial location of the labelling groups;
- and
- vii. expanding the hydrogel network by promoting the dissociation of the electrolyte monomer by water exchange.
A method for enlarging a sample of interest comprising (i) providing a sample of interest permeated with a swellable, photopolymerizable hydrogel network;
- (ii) cross-linking the swellable, photopolymerizable hydrogel network through the application of light irradiation wherein the control of its crosslinking density is applied by varying the light irradiation conditions;
- and (iii) swelling the cross-linked hydrogel network to physically enlarge the sample of interest.
The method according to claim 25 wherein the hydrogel is prepared via thiol-acrylate photopolymerization and the hydrogel formulation is composed of a photoinitiator, a crosslinking monomer, an electrolyte monomer, a chain transfer agent and a plasticizing monomer.
Dependencies are inferred from the claim text.