Structure for introducing a plurality of solutions, micro fluidic device having said structure and method for introducing solution

1. A method for separating a complex between an analyte or an analogue thereof in a sample, and a substance binding to said analyte or analogue thereof, which comprises:

(a) providing a micro fluidic device comprising at least one structure for introducing a sample or a reagent solution to a first channel ( 1 ) in a substrate, said structure comprising;

(i) the first channel ( 1 ), said first channel ( 1 ) having an upstream reservoir ( 6 ) at the end of the upstream side thereof and a downstream reservoir (7) at the other end of the downstream side thereof,

(ii) not less than 3 side channels ( 2 ) for drain, each said side channel ( 2 ) for drain having a opening ( 2 - 1 ) opened on a wall of said first channel ( 1 ), and said openings ( 2 - 1 ) arranged in spaced relation to one another, wherein said side channel ( 2 ) for drain is connected to a waste reservoir ( 4 );

(iii) not less than 2 side channels ( 3 ) for introducing a sample or a reagent solution, each said side channel ( 3 ) for introducing a sample or a reagent solution having a opening ( 3 - 1 ) opened on the wall of said first channel ( 1 ) at the different position (part) from said openings ( 2 - 1 ) of said side channels ( 2 ) for drain, and each said opening ( 3 - 1 ) arranged between adjacent 2 openings ( 2 - 1 ) of side channels ( 2 ) for drain, wherein each said opening ( 3 - 1 ) of these not less than 2 side channels ( 3 ) for introducing a sample or a reagent solution are each arranged at the different portion between adjacent 2 openings ( 2 - 1 ) of side channels ( 2 ) for drain, and side channel ( 3 ) for introducing a sample or a reagent solution is connected to a sample or a reagent solution reservoir ( 5 );

(iv) a side channel ( 8 ) for introducing an electrophoresis medium communicating with the first channel region between a side channel ( 2 n ) for drain located at the most downstream among said not less than 3 side channels ( 2 ) for drain and a downstream reservoir ( 7 ), and said side channel ( 8 ) for introducing an electrophoresis medium connecting to an electrophoresis medium reservoir ( 11 );

(b) putting a sample into at least one sample or reagent solution reservoir (5) connecting to said side channel ( 3 ) for introducing a sample or a reagent solution;

(c) putting a reagent solution into at least one sample or reagent solution reservoir ( 5 ) connecting to said side channel ( 3 ) for introducing a sample or a reagent solution, wherein said sample or reagent reservoir ( 5 ) being different from one used in said step (b);

(d) introducing said sample in said side channel ( 3 ) for introducing a sample or a reagent solution connected with said sample or reagent solution reservoir ( 5 ) used in said step (b), 2 side channels ( 2 ) for drain adjacent to said side channel ( 3 ) for introducing a sample or a reagent solution in an axis direction and a first channel region between said 2 side channels ( 2 ) for drain, by moving said sample from said sample or reagent solution reservoir ( 5 ) where said sample is put in said step (b) to said 2 side channels ( 2 ) for drain adjacent to said side channel ( 3 ) for introducing a sample or a reagent solution connected with said sample or reagent solution reservoir ( 5 ) in an axis direction, via said side channel ( 3 ) for introducing a sample or a reagent solution connected with said reservoir ( 5 );

(e) introducing said reagent solution in said side channel ( 3 ) for introducing a sample or a reagent solution connected with said sample or reagent solution reservoir ( 5 ) used in said step (c), 2 side channels ( 2 ) for drain adjacent to said side channel ( 3 ) for introducing a sample or a reagent solution in an axis direction and a first channel region between said 2 side channels ( 2 ) for drain, by moving said reagent solution from said sample or reagent solution reservoir ( 5 ) where said reagent solution is put in said step (c) to said 2 side channels ( 2 ) for drain adjacent to said side channel ( 3 ) for introducing a sample or a reagent solution connected with said sample or reagent solution reservoir ( 5 ) in an axis direction, via said side channel ( 3 ) for introducing a sample or a reagent solution connected with said reservoir ( 5 );

(f) reacting an analyte or an analogue thereof in a sample with at least one kind of a substances binding to said analyte or analogue thereof while concentrating said analyte or said analogue thereof in the sample and/or at least one kind of the substances binding to said analyte or analogue thereof in the reagent solution to form the complex between said analyte or said analogue thereof and the substances binding to said analyte or analogue thereof, at a first channel region ( 9 a ) between a side channel ( 2 a ) for drain located at the most upstream among said not less than 3 side channels ( 2 ) for drain and a side channel ( 2 n ) for drain located at the most downstream among said not less than 3 side channels ( 2 ) for drain, or a first channel region ( 9 b ) between a side channel ( 2 a ) for drain located at the most upstream among said not less than 3 side channels ( 2 ) for drain and said side channel ( 8 ) for introducing an electrophoresis medium by applying a voltage to said upstream reservoir ( 6 ) part at the end of said upstream side of said first channel ( 1 ) and said downstream reservoir ( 7 ) part at the other end of said downstream side of said first channel ( 1 ); and

(g) separating said complex and said substance binding to said analyte not involved in the formation of said complex or said analogue not involved in the formation of said complex, by applying a voltage to said electrophoresis medium reservoir ( 11 ) part connecting to said side channel ( 8 ) for introducing an electrophoresis medium and said downstream reservoir ( 7 ) part.

2. A method for measuring an analyte or an analogue thereof in a sample, which comprises:

(a) providing a micro fluidic device comprising at least one structure for introducing a sample or a reagent solution to a first channel ( 1 ) in a substrate, said structure comprising;

(i) the first channel ( 1 ), said first channel ( 1 ) having an upstream reservoir ( 6 ) at the end of the upstream side thereof and a downstream reservoir ( 7 ) at the other end of the downstream side thereof,

(ii) not less than 3 side channels ( 2 ) for drain, each said side channel ( 2 ) for drain having a opening ( 2 - 1 ) opened on a wall of said first channel ( 1 ), and said openings ( 2 - 1 ) arranged in spaced relation to one another, wherein said side channel ( 2 ) for drain is connected to a waste reservoir ( 4 );

(iii) not less than 2 side channels ( 3 ) for introducing a sample or a reagent solution, each said side channel ( 3 ) for introducing a sample or a reagent solution having a opening ( 3 - 1 ) opened on the wall of said first channel ( 1 ) at the different position (part) from said openings ( 2 - 1 ) of said side channels ( 2 ) for drain, and each said opening ( 3 - 1 ) arranged between adjacent 2 openings ( 2 - 1 ) of side channels ( 2 ) for drain, wherein each said opening ( 3 - 1 ) of these not less than 2 side channels ( 3 ) for introducing a sample or a reagent solution are each arranged at the different portion between adjacent 2 openings ( 2 - 1 ) of side channels ( 2 ) for drain, and side channel ( 3 ) for introducing a sample or a reagent solution is connected to a sample or a reagent solution reservoir ( 5 );

(iv) a side channel ( 8 ) for introducing an electrophoresis medium, said side channel ( 8 ) for introducing an electrophoresis medium communicating with the first channel region between a side channel ( 2 n ) for drain located at the most downstream among said not less than 3 side channels ( 2 ) for drain and a downstream reservoir ( 7 ), and said side channel ( 8 ) for introducing an electrophoresis medium connecting to an electrophoresis medium reservoir ( 11 );

(b) putting a sample into at least one sample or reagent solution reservoir ( 5 ) connecting to said side channel ( 3 ) for introducing a sample or a reagent solution;

(c) putting a reagent solution into at least one sample or reagent solution reservoir ( 5 ) connecting to said side channel ( 3 ) for introducing a sample or a reagent solution, wherein said sample or reagent reservoir ( 5 ) being different from one used in said step (b);

(d) introducing said sample in said side channel ( 3 ) for introducing a sample or a reagent solution connected with said sample or reagent solution reservoir ( 5 ) used in said step (b), 2 side channels ( 2 ) for drain adjacent to said side channel ( 3 ) for introducing a sample or a reagent solution in an axis direction and a first channel region between said 2 side channels ( 2 ) for drain, by moving said sample from said sample or reagent solution reservoir ( 5 ) where said sample is put in said step (b) to said 2 side channels ( 2 ) for drain adjacent to said side channel ( 3 ) for introducing a sample or a reagent solution connected with said sample or reagent solution reservoir ( 5 ) in an axis direction, via said side channel ( 3 ) for introducing a sample or a reagent solution connected with said reservoir ( 5 );

(e) introducing said reagent solution in said side channel ( 3 ) for introducing a sample or a reagent solution connected with said sample or reagent solution reservoir ( 5 ) used in said step (c), 2 side channels ( 2 ) for drain adjacent to said side channel ( 3 ) for introducing a sample or a reagent solution in an axis direction and a first channel region between said 2 side channels ( 2 ) for drain, by moving said reagent solution from said sample or reagent solution reservoir ( 5 ) where said reagent solution is put in said step (c) to said 2 side channels ( 2 ) for drain adjacent to said side channel ( 3 ) for introducing a sample or a reagent solution connected with said sample or reagent solution reservoir ( 5 ) in an axis direction, via said side channel ( 3 ) for introducing a sample or a reagent solution connected with said reservoir ( 5 );

(f) reacting an analyte or an analogue thereof in a sample with at least one kind of a substances binding to said analyte or analogue thereof while concentrating said analyte or said analogue thereof in the sample and/or at least one kind of the substances binding to said analyte or analogue thereof in the reagent solution to form the complex between said analyte or said analogue thereof and the substances binding to said analyte or analogue thereof, at a first channel region ( 9 a ) between a side channel ( 2 a ) for drain located at the most upstream among said not less than 3 side channels ( 2 ) for drain and a side channel ( 2 n ) for drain located at the most downstream among said not less than 3 side channels ( 2 ) for drain, or a first channel region ( 9 b ) between a side channel ( 2 a ) for drain located at the most upstream among said not less than 3 side channels ( 2 ) for drain and said side channel ( 8 ) for introducing an electrophoresis medium by applying a voltage to said upstream reservoir ( 6 ) part at the end of said upstream side of said first channel ( 1 ) and said downstream reservoir ( 7 ) part at the other end of said downstream side of said first channel ( 1 );

(g) separating said complex and said substance binding to said analyte or said analogue thereof not involved in the formation of said complex, or said analogue not involved in the formation of said complex, by applying a voltage to said electrophoresis medium reservoir ( 11 ) part connecting to said side channel ( 8 ) for introducing an electrophoresis medium and said downstream reservoir ( 7 ) part; and

(h) measuring the amount of thus separated complex, the amount of said substance binding to said analyte or said analogue not involved in the formation of said complex, or the amount of said analogue not involved in the formation of said complex to determine the amount of said analyte or said analogue thereof in a sample based on the result.