Embedding medium for biological samples, method for producing embedded biological samples, and use thereof

Patent metadata

Field	Value
Publication number	US-10401266-B2
Application number	US-201515319611-A
Country	US
Kind code	B2
Filing date	Jun 19, 2015
Priority date	Jun 19, 2014
Publication date	Sep 3, 2019
Grant date	Sep 3, 2019

Abstract

In a first aspect, the invention relates to the use of a UV-polymerizable composition as an embedding medium for biological samples. The UV-polymerizable composition is a composition with a refractive index ranging from n=1.45 to n=1.6 after a polymerization process. In another aspect, the invention relates to a method for producing embedded biological samples using said UV-polymerizable composition and to the embedded biological samples themselves. The biological samples can be used in a wide variety of areas, such as diagnostics among others, and are suitable in particular for RNA-based diagnostics.

First claim

The invention claimed is:

1. A method for producing an embedded biological sample, comprising the steps:
a) dewatering the biological sample,
b) infusing a UV-polymerizable composition into the biological sample, wherein said UV-polymerizable composition has a refractive index after polymerization in the range from n=1.45 to n=1.6,
c) embedding the infused biological sample by curing the UV-polymerizable composition with light having a wavelength of ≤470 nm, and the UV-polymerizable composition comprises a mercapto ester compound.

2. The method as claimed in claim 1 , whereby polymerization does not proceed continuously at the start.

3. The method as claimed in claim 1 , wherein the biological sample was fixed before the dewatering.

4. The method as claimed in claim 1 , wherein at least the infusion of the UV-polymerizable composition is carried out under vacuum.

5. The method as claimed in claim 1 , wherein the dewatering of the biological sample proceeds i) by means of an ascending alcohol series or ii) by means of an ascending DMSO series.

6. The method as claimed claim 1 , wherein the mercapto ester compound is present in the UV-polymerizable composition at at least 40% by weight.

7. The method as claimed in claim 1 , wherein the refractive index of the UV-polymerizable composition before polymerization is in a range from n=1.4 to 1.6.

8. The method as claimed in claim 1 , wherein the UV-polymerizable composition additionally comprises at least one of a polymerizable acrylate compound and a methacrylate compound.

9. The method as claimed in claim 1 , wherein the biological sample is a three-dimensional biological sample and wherein said three-dimensional sample has a smallest edge length of at least 50 μm.

10. The method according to claim 1 wherein said UV-polymerizable composition has a refractive index after polymerization in the range from n=1.550 to n=1.565.

11. The method according to claim 2 further comprising cooling the biological sample between incidences of light having a wavelength of ≤470 nm.

12. The method as claimed in claim 3 wherein the biological sample was fixed with aldehyde mixtures or alcohols.

13. The method as claimed in claim 5 , wherein the dewatering of the biological sample proceeds by means of an ascending ethanol series.

14. The method according to claim 5 further comprising subsequent dewatering in xylene.

15. The method as claimed in claim 7 wherein the refractive index of the UV-polymerizable composition before polymerization is in the range from n=1.50 to 1.55.

16. The method as claimed in claim 9 wherein the biological sample is a three-dimensional biological sample and wherein said three-dimensional sample has a smallest edge length of at least 100 μm.

Assignees

• Medizinische Hochschule Hannover
• Hannover Laser Zentrum

Inventors

• Kuehnel Mark
• Kellner Manuela
• Jonigk Danny
• Izykowski Nicole
• Meyer Heiko
• Lorbeer Raoul-Amadeus
• Heidrich Marko

Classifications

• G01N2001/364

  using resins, epoxy · CPC title

• G01N1/30 Primary

  Staining; Impregnating {; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis} · CPC title

• G01N1/44

  Sample treatment involving radiation, e.g. heat · CPC title

• G01N2001/305

  Fixative compositions · CPC title

• G01N1/286

  involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising (microtomes G01N1/06; pulverising in general B02C; mixing in general B01F) · CPC title