Method and apparatus for identifying and separating a specific target component from multiple components.

Abstract

The present invention relates to a method for identifying and separating a specific component of interest from a plurality of components, the method comprising: A) preparing a dispersion comprising the plurality of components, which may be encapsulated in compartments, and a photoresist, wherein at least a portion of the plurality of components exhibits a factor of interest; B) scanning the dispersion for at least one factor of interest to identify the plurality of components or optionally at least a portion of the compartments, selecting these components of interest or optionally compartments, and assigning locations to the components of interest or optionally compartments within the dispersion; C) irradiating at least a portion of the dispersion with light to cure the photoresist, which may or may not be attributed to the component of interest; and D) separating components in the portion of the dispersion where the photoresist has been cured from components in the other portion of the dispersion where the photoresist has not been cured. Furthermore, the present invention relates to an apparatus configured to perform the method according to any one of the above claims.

Claims

1. A method for identifying and separating a specific component from multiple components, A) A step of preparing a dispersion comprising the above-mentioned plurality of components, which may be encapsulated in compartments, and a photoresist, wherein at least a portion of the above-mentioned plurality of components exhibits the target factor. B) A step of scanning the dispersion for at least one target factor to identify at least a portion of the multiple components or optionally the compartments, selecting these target components or optionally the compartments, and assigning a location to the target component or optionally the compartments within the dispersion. C) A step of curing the photoresist which belongs to or does not belong to the target component by irradiating at least a portion of the dispersion with light, and D) A step of separating the components in the portion of the dispersion in which the photoresist has been cured from the components in the other portions of the dispersion in which the photoresist has not been cured. A method that includes this.

2. The method according to claim 1, wherein the above compartment is one of the following: a droplet containing the above component, preferably a water-in-oil droplet or a double emulsion droplet, a vesicle, preferably a multilayer vesicle, a monolayer vesicle or GUV, or a coacervate.

3. The method according to claim 1 or 2, wherein the above component is a natural cell, a synthetic cell, an organelle, a protein, a nucleic acid, or an inorganic particle, and preferably the above component is a living cell.

4. The method according to claim 1 or 2, wherein the component for the above purpose or optionally the compartment for the purpose differs from other components or optionally other compartments by optical characteristics, for example, the fluorescent portion and / or the shape and / or behavior of the component.

5. The method according to claim 1 or 2, wherein the photoresist is a negative-type photoresist and comprises a monomer component and an initiator component.

6. The method according to claim 1 or 2, wherein the photoresist is a biocompatible photoresist.

7. The method according to claim 1 or 2, wherein the light is an LED or a laser beam, and in particular, the light is focused light.

8. The method according to claim 1 or 2, wherein in step C), the light is configured to selectively irradiate at least a portion of the dispersion.

9. In step A), the dispersion is prepared in a chamber having a row of subchambers, each designed to incorporate one or more components, preferably the chamber is designed to have only one layer of components which may be enclosed in compartments, and / or The method according to claim 1 or 2, wherein the concentration of the component which may be sealed in the compartment in step A) is adjusted so that a single layer of the component which may be sealed in the compartment is formed in the chamber.

10. The method according to claim 9, further comprising step B0) before step B), wherein B0) is to scan the chamber to determine a suitable depth of field for a camera observing the chamber.

11. The above method is carried out using a microscope having a light source suitable for polymerizing the photoresist, and / or The method according to claim 1 or 2, wherein the above method is performed using a microscope having a camera, and in step B), an image is captured by the camera and analyzed, in particular by a computer, for at least one factor of interest.

12. The method according to claim 1, wherein the above method is performed using a microscope that provides structured illumination, and preferably the microscope has a digital micromirror device that allows the curing of the photoresist in step C) to be performed in parallel at several locations.

13. The above method is partially performed by a computer, and in particular, the above computer-based method is: In optional step B0), the scanning of the chamber is configured to determine the depth of focus for capturing an image of the chamber. In step B), an image is acquired, the image is analyzed for at least one target factor, and the location is assigned to the target component in the dispersion, and In step C), the light source is controlled to selectively illuminate the photoresist in the desired area to cure the photoresist. The method according to claim 1, 2, or 12, including the following:

14. Process B) and / or process C) are performed automatically, and / or based on machine learning-based image analysis. In step B), the acquisition of the above image is controlled by the computer analyzing the above image and adjusting the focus and / or coordinates of the captured image, and / or The method according to claim 1, 2, or 12, wherein steps B) and C) are repeated multiple times.

15. An apparatus configured to perform the method according to claim 1, 2, or 12, preferably including a microscope.

16. The use of a photoresist for cell sorting, particularly for sorting live cells, wherein the photoresist comprises a monomer component and an initiator component. The above monomer component is a photoresist having a chemical structure that includes polymerizable groups bonded to cell-impermeable groups.

17. The use of the photoresist according to claim 16, wherein the impermeable group is a residue of an impermeable molecule selected from the group consisting of sugars such as glucose, sulfur oxides, carboxylic acids, and combinations thereof.

18. The use of the photoresist according to claim 16, wherein the impermeable group is an impermeable polymer fragment based on a polymer selected from the group consisting of alginate, dextran, hyaluronic acid, and / or multi-arm polyethylene glycol derivatives (PEG).

19. The use of the photoresist according to claim 16, wherein the monomer component is dextranglycidyl methacrylate.

20. The use of a photoresist according to any one of claims 16 to 19, wherein the polymerizable group is selected from the group consisting of an acrylate group, a methacrylate group, an acrylamide group, or a methacrylamide group.

21. Use of a photoresist according to any one of claims 16 to 19 for sorting living cells using the method according to claim 1, 2, or 12, particularly the method according to claim 12.

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