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US 12,656,231 B2Granted
Refractive index matching formulations
AssigneeLife Technologies Corporation
InventorsAdam York, Eric Welch, Daniel Cash
Life of this patent
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Abstract
Refractive index matching (RIM) formulations, their use for mounting biological specimens (e.g., cells and tissues) to a substrate, and methods of visualizing biological specimens embedded in RIM formulations are described.
Claims17
A refractive index matching (RIM) solution, comprising:
- a) a water-soluble polymer, wherein the water-soluble polymer is (i) a charged water-soluble polymer;
- (ii) an uncharged, water-soluble polymer that comprises a monomer residue of N—R acrylamide or N—R methacrylamide, wherein R is methyl, ethyl, propyl, isopropyl or H, N,N-dimethylacrylamide, N,N-dimethylmethacrylamide, N,N-diethylacrylamide, N, N-diethylmethacrylamide, N-2-hydroxypropyl methacrylamide, or a combination thereof;
- or (iii) poly(methyl vinyl ether), poly(2-ethyl-2-oxazoline), poly(2-methyl-2-oxazoline), poly(vinyl pyrrolidone) (PVP), or polyvinyl alcohol (PVA);
- b) a polyol, wherein the polyol is polyvinyl alcohol, glycerol, diglycerol, polyglycerol, mannitol, sorbitol, erythritol, thiodiethanol, thiodipropanol, or a combination thereof, and wherein the weight ratio of the water-soluble polymer to the polyol is 0.125:1 to 4:1, wherein the water-soluble polymer and polyol are different;
- and
- c) water or a buffer, wherein the RIM solution has a refractive index of about 1.36 to about 1.60.
3 more elements
A method of mounting a biological specimen on a substrate, comprising:
- depositing the biological specimen on the substrate;
- contacting the biological specimen with a refractive index matching (RIM) solution to provide a mounted sample, wherein the RIM solution comprises:
- a) a water-soluble polymer, wherein the water-soluble polymer is (i) a charged water-soluble polymer;
- (ii) an uncharged, water-soluble polymer that comprises a monomer residue of N—R acrylamide or N—R methacrylamide, wherein R is methyl, ethyl, propyl, isopropyl or H, N,N-dimethylacrylamide, N,N-dimethylmethacrylamide, N,N-diethylacrylamide, N, N-diethylmethacrylamide, N-2-hydroxypropyl methacrylamide, or a combination thereof;
- or (iii) poly(methyl vinyl ether), poly(2-ethyl-2-oxazoline), poly(2-methyl-2-oxazoline), poly(vinyl pyrrolidone) (PVP), or polyvinyl alcohol (PVA),
- b) a polyol, wherein the weight ratio of the water-soluble polymer to the polyol is 0.125:1 to 4:1, wherein the water-soluble polymer and polyol are different, and
- c) water or a buffer, wherein the RIM solution has a refractive index of about 1.36 to about 1.60;
- and
- drying the mounted sample at room temperature to about 40° C. for about 48 hours or less to provide a solidified mountant.
6 more elements
A solidified mountant, comprising a water-soluble polymer, and a polyol, wherein the weight ratio of the water-soluble polymer to the polyol is 0.125:1 to 4:1, wherein the water-soluble polymer is (i) a charged water-soluble polymer;
- (ii) an uncharged, water-soluble polymer that comprises a monomer residue of N—R acrylamide or N—R methacrylamide, wherein R is methyl, ethyl, propyl, isopropyl or H, N,N-dimethylacrylamide, N,N-dimethylmethacrylamide, N,N-diethylacrylamide, N, N-diethylmethacrylamide, N-2-hydroxypropyl methacrylamide, or a combination thereof;
- or (iii) poly(methyl vinyl ether), poly(2-ethyl-2-oxazoline), poly(2-methyl-2-oxazoline), poly(vinyl pyrrolidone) (PVP), or polyvinyl alcohol (PVA), wherein the solidified mountant has a refractive index of about 1.48 to about 1.60 and wherein the water-soluble polymer and polyol are different.
Dependencies are inferred from the claim text.