Method to distribute liquids containing molecules in solution and to deposit said molecules on solid supports, and relative device
OwnerGiuseppe Firraoinventor
Life of this patent
Estimating the term…
Abstract
Method and device to distribute at least a sample of liquid containing molecules in solution, and to deposit and bind the molecules to solid supports. The method provides to use at least two supports (12; 112) which have been previously equipped with at least a through hole (11) and to arrange them one on top of the other so that the respective holes (11) are aligned so as to define a containing channel (13). The method then provides to distribute the sample of liquid inside the containing channel (13) using a surface inside the holes (11) as a site for depositing, localizing and binding the molecules in solution in the sample of liquid.
Claims41
1- (Amended) Method to distribute at least a sample of liquid containing molecules in solution, and to deposit and bind said molecules to solid supports, characterized in that it provides to use at least two supports (12;
- 112) which
- have been previously equipped with at least two through holes (11) spaced but extremely close each other and
- have been previously arranged in direct contact one on top of the other
- said two supports (12;
- 112) being aligned so that the respective holes (11) define a containing channel (13), the method further providing to distribute said at least one sample of liquid inside said containing channel (13) using a surface inside said holes (11) as a direct site to deposit, localize and bind the molecules in solution in said sample of liquid,
- in which, having terminated the step wherein the molecules in solution are bound to the inner surface of the hole (11) of the support (12;
- 112), said supports (12;
- 112) are separated from each other, each defining a collection of. immobilized molecules for use in experiments and tests for diagnosis and research.
2- (New) Method as in any claim hereinbefore, characterized in that said through holes (11) are spaced each other about 1,5 mm or less.
3- Method as in claim 1 or 2, characterized in that the surface on which said molecules in solution in said sample of liquid are deposited, localized and bound forms an angle of less than 90° with the surface of said supports (12;
- 112) on which said holes (11) have been made.
4- Method as in claim 1 or 2, characterized in that each of said supports (12;
- 112) consists of a central layer (19) coated on at least one face with at least a separating layer (20) of different material.
5- Method as in claim 4, characterized in that said hole (11) is formed by the separating layers (20) and is partly obstructed at the center by the central layer (19) in the form of a grid which constitutes the surface on which said molecules in solution in said sample of liquid are deposited, localized and bound.
6- Method as in any claim hereinbefore, characterized in that said surface on which said molecules in solution in said sample of liquid are deposited, localized and bound is coated with a material suitable for allowing or improving the binding of said molecules.
7- Method as in claim 6, characterized in that said material is localized exclusively on said central layer (19) and not on said separating layer (20).
8- Method as in claim 1, characterized in that after the molecules have been bound to said inner surface of the holes (11), it provides that said at least two supports (12) are separated from each other to be used in independent manner.
9- Method as in any claim hereinbefore, characterized in that said molecules consist of polymers of biological interest, or biomolecules.
10- Method as in claim 9, characterized in that said biomolecules are nucleic acids.
11- Method as in claim 9, characterized in that said biomolecules are proteins.
12- Method as in claim 9, characterized in that said biomolecules are substituted by functional groups.
13- Method as in claim 1, characterized in that it provides to use at least selective means (18a;
- 119) arranged below the support (12;112) located at the bottom, to close said containing channel (13) for the sample of liquid defined by the aligned through holes (11) of said at least two supports (12;
- 112).
14- Method as in claim 13, characterized in that said means (18a;
- 119) consist of at least an element made of hydrophobic material.
15- Method as in claim 13, characterized in that said means (18a;
- 119) comprise at least a porous membrane.
16- Method as in claim 13, characterized in that it provides at least a step of applying vacuum at least from below said closing means (18a;
- 119) to obtain a desired filling of the containing channel (13) defined by said aligned through holes (11) of said at least two supports (12;
- 112).
17- Method as in claim 1, characterized in that when said containing channel (13) has been filled with said sample of liquid, it provides that said containing channel (13) is sealed to allow the binding of the biomolecules to the inner surfaces of the hole (11) of the respective support (12;
- 112).
18- Method as in claim 17, characterized in that it provides that, after sealing, the liquid inside said containing channel is subjected to heat, electric or other physical treatment in order to obtain the binding of the biomolecules to the surfaces inside the hole (11) of the respective support (12;
- 112).
19- Method as in claim 1, characterized in that when said containing channel (13) has been filled with said sample of liquid, it provides to apply an electric potential in order to obtain the binding of the biomolecules to the surfaces inside the hole (11) of the respective support (12;
- 112).
20- Method as in any claim hereinbefore, characterized in that after the containing channel (13) has been filled, it provides that said supports (12;
- 112) are arranged in an environment at a controlled temperature to promote the binding of the biomolecules on the inner wall of said through holes (11).
21- Method as in any claim hereinbefore, characterized in that it provides that said containing channels (13) are filled sequentially in manual mode.
22- Method as in any claims from 1 to 20 inclusive, characterized in that it provides that a plurality of said containing channels (13) are filled simultaneously and automatically by means of suitable distributor means (52, 53, 55).
23- Method as in claim 22, characterized in that said simultaneous filling of a plurality of containing channels (13) occurs with a single movement of said distributor means (52, 53, 55).
24- Device to distribute and deposit on solid supports at least a sample of liquid containing molecules in solution, characterized in that it comprises at least two supports (12;
- 112) which have been already previously equipped with at least two through holes (11) spaced but very close each other, by a distance of 1,5 mm or less, which may be partly obstructed, and arranged one on top of the other so that the respective holes (11) are aligned so as to define a containing channel (13), means (18a;
- 119) arranged below the support (12;
- 112) located at the bottom to close said containing channel (13), and means able to distribute said at least one sample of liquid inside said containing channel (13) using a surface inside said holes (11) as a direct site to deposit, localize and bind the molecules in solution in said sample of liquid.
25- Device as in claim 24, characterized in that it comprises means (14;
- 114) able to create a vacuum associated at least with below said supports (12;
- 112).
26- Device as in claim 24, characterized in that it comprises at least a lower base plate (18;
- 118) and at least an upper base plate (18b;
- 120), said plates (18, 18b;
- 118, 120) including respective holes (29, 34) in correspondence with the holes (11) made on the supports (12;
- 112) and able to enclose said superimposed and aligned supports (12) and said closing means (18a) between them.
27- Device as in claim 26, characterized in that said elements (12, 18, 18a, 18b;
- 112, 118, 119, 120) are able to be constrained one to the other by means of joining means (16;
- 116), forming in their entirety a rigid block (15;
- 115) able to be associated at least temporarily to said means (14;
- 114) able to create a vacuum to distribute the samples of liquid inside the holes (11) of said supports (12;
- 112).
28- Device as in claim 25, characterized in that said means able to create a vacuum comprise at least a chamber (14;
- 114) including at least a first aperture (21) for connection with pump means able to create a vacuum and at least a second window-type aperture (23) for positioning said rigid block (15;
- 115) by means of clamping means (17).
29- Device as in claim 28, characterized in that sealing means (25) are interposed between said second window-type aperture (23) and said rigid block (15;
- 115).
30- Device as in any claims from 24 to 29 inclusive, characterized in that each of said supports (12;
- 112) has a thickness comprised between about 0.15 and 0.45 mm.
31- Device as in any claims from 24 to 30 inclusive, characterized in that said through holes (11) made on said supports (12;
- 112) have a maximum diameter of about 1 mm.
32- Device as in any claims from 27 to 31 inclusive, characterized in that it comprises assembly means (36;
- 136a, 136b) able to facilitate the assembly of said rigid block (15;
- 115).
33- Device as in claim 32, characterized in that said assembly means (36;
- 136a, 136b) comprise guide elements (39;
- 43, 44) able to cooperate with mating holes (28, 31, 35) made respectively in said elements (18, 12, 18b;
- 118, 112, 120).
34- Device as in any claims from 24 to 33 inclusive, characterized in that it comprises automatic distribution means for said samples of liquid containing biomolecules in solution inside said holes (11) made on said supports (12;
- 112).
35- Device as in claim 34, characterized in that said automatic distribution means comprise at least plate means (52) able to move at least on a plane parallel to the plane on which said rigid block (15;
- 115) lies, said plate means (52) including through holes (56) able to allow said samples of liquid to pass.
36- Device as in claim 35, characterized in that said through holes (56) have a substantially conical conformation.
37- Device as in claim 35, characterized in that said plate means (52) are associated with cylinder-type movement means (53) and thrust means (55) to transmit motion.
38- Device as in claim 35, characterized in that said movable plate means (52) and said rigid block (15;
- 115) comprising said supports (12) are equipped with a reciprocal vertical movement to be taken into reciprocal contact at least during the step of unloading the samples of liquid from the holes (56) of said movable plate (52) to the holes (11) of said supports (12).
39- Device as in claim 38, characterized in that at least said rigid block (15;
- 115) is mounted on a plane (47) movable at least in height.
40- Device as in claim 35, characterized in that it comprises washing means (57) able to perform a washing cycle to wash the holes (56) of said movable plate means (52) after every cycle to distribute said samples of liquid.
41- Device as in claim 35, characterized in that the number of holes (56) on said movable plate means (52) is equal to a sub-multiple of the number of holes (11) on said supports (12).