Swellable photopolymerized hydrogels for expansion microscopy
AssigneeThe Regents of the University of Colorado, a body corporate
InventorsKristi Anseth, Kemal Arda Gunay, Laura J. Macdougall
Life of this patent
Estimating the term…
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.
Claims10
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 a 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 electrolyte monomers by water exchange;
- wherein the labelling groups are applied prior to the digesting step and remain associated with the tethered biological cue throughout the digesting step and the expanding step, without removal and re-application of the labelling groups after digestion.
Dependencies are inferred from the claim text.