WO2008143415A1

WO2008143415A1 - Lab-on-a-chip comprising a mixing chamber

Info

Publication number: WO2008143415A1
Application number: PCT/KR2008/002561
Authority: WO
Inventors: Ji Hoon Lee; Young Teck Kim; Hee Sang Suh
Original assignee: Lst Korea. Inc.
Priority date: 2007-05-17
Filing date: 2008-05-07
Publication date: 2008-11-27
Also published as: KR100864635B1

Abstract

A lab-on-a-chip 1 comprising a mixing chamber comprises a first panel 10 and a second panel 30, which are formed of a transparent material, the first panel 10 including: a reagent injection part 11 formed to inject a reagent; a sample injection part 13 formed to inject a sample; a reagent channel 15 extended from the reagent injection part 11 at a predetermined width; a sample injection channel 17 extended from the sample injection part 13 at a predetermined width; a mixing chamber 19 formed at a place where the reagent channel 15 meets the sample channel 17, mixing a reagent with a sample; a mixing channel 21 extended from the mixing chamber 19 at a predetermined angle with respect to the reagent channel 15 and the sample channel 17, and a detection part 14 formed at the end of the mixing channel 21, and the second panel 30 including: a reagent injection hole 31 which is a through hole connected to the reagent injection part 11; a sample injection hole 33 which is a through hole connected to the sample injection part 13; and an exhaust hole 35 which is a through hole connected to the detection part 14, wherein the mixing chamber 19 is wider than the reagent channel 15 or the sample channel 17. In this case, the sample is mixed with the reagent uniformly and promptly without a narrow and complicate mixing channel which is difficult to be processed. The low concentration of sample was quantified with excellent reproducibility within statistical error range using the lab-on-a-chip, 1 with chemiluminescence detection.

Description

Description LAB-ON-A-CHIP COMPRISING A MIXING CHAMBER

Technical Field

[1] The present invention relates to a lab-on-a-chip, and more particularly, to a lab- on-a-chip comprising a mixing chamber at a part where injection channels of a sample and a reagent meet each other, so as to mix the sample with the reagent uniformly and promptly. Background Art

[2] A lab-on-a chip is a chemical microprocessor in which various devices are integrated on a substrate having a size of several cm² using micromachining technique such as photolithography technique used in a process of manufacturing a semiconductor, wherein the substrate is made of glass, silicon, or plastic. An automated experiment of high speed, high efficiency, and low cost can be carried out through the lab- on-a-chip(see reference [Kovacs, Anal. Chem. 68 (1996) 407A-412A]).

[3] FIG. 1 is a plane view illustrating a lab-on-a-chip according to the related art. As shown in FIG. 1, a lab-on-a-chip 100 according to the related art comprises a luminescence zone 130 comprised of a fine channel 131 which induces mixture of a sample and an enzyme mixture (reagent) in a transparent plate material.

[4] The lab-on-a-chip 100 shown in FIG. 1 comprises a reagent injection part 121 injecting an enzyme mixture (reagent), a sample injection part 123 injecting a sample to be measured, a reagent channel 125 extended from the reagent injection part 121 at a predetermined width, a sample channel 127 extended from the sample injection part 123 at a predetermined width and formed in parallel with the reagent channel 125 within a predetermined range, and a luminescence zone 130 formed at the downstream of a mixing channel 141 formed at a place where the reagent channel 125 meets the sample channel 127. The mixing channel 141 is extended to the downstream of the luminescence zone 130, and a mixing solution exhaust part 143 is formed at the end of the mixing channel 141.

[5] The luminescence zone 130 is comprised of a fine channel 131 which is formed by reciprocation of a capillary tube in left and right directions, wherein the capillary tube has a width smaller than that of the reagent channel 125 or the sample channel 127. Since the fine channel 131 constituting the luminescence zone 130 is longer than the other channels 125, 127 and 141, it stays in the luminescence zone 130 for a long time to enable an excellent luminescence function, as disclosed in Laid-Open Publication (Patent Publication 10-2006-102794). A reference numeral 110 of FIG. 1, which is not described, depicts a panel constituting the lab-on-a-chip 100.

[6] FIG. 2 is a plane view illustrating another light-emitting lab-on-a-chip according to the related art. As shown in FIG. 2, a lab-on-a-chip 200 according to the related art includes a reagent injection part 221 injecting a reaction solution, a sample injection part 223 injecting a sample to be measured, a reagent channel 225 extended from the reagent injection part 221 at a predetermined width, a sample channel 227 extended from the sample injection part 223 at a predetermined width, and a column reactor 230 formed at the downstream of a mixing channel formed at a place where the reagent channel 225 meets the sample channel 227. A reaction channel 231 of the column reactor 230 is formed to be bend toward a fine tube, so as to enable prompt mixture and reaction of a sample solution and a reaction solution at high efficiency. In FIG. 2, a reference numeral 210 depicts a panel where the channels are formed, a reference numeral 251 depicts a buffer solution vessel, a reference numeral 253 depicts a reaction solution exhaust part, and a reference numeral 241 depicts a channel connected to a buffer solution exhaust part. Disclosure of Invention Technical Problem

[7] Fbwever, the aforementioned lab-on-a-chips 100 and 200 according to the related art have been developed based on a mixing principle by diffusion such as a diffusion coefficient of reactants and a laminar flow effect. In order to mix the reactants by using the above principle, a fine tube which is narrow and long is required. In other words, in order to mix a reagent (or reaction solution) with a sample and detect luminescence generated by mixing, the fine channel 131 or the reaction channel 231 should be provided, wherein the fine channel 131 is formed by repetition of several times in a vertical direction, and the reactoin channel 231 is formed by a fine tube bent several times. For this reason, it is difficult to manufacture the channel. This results in much time to manufacture the channel. Also, a problem occurs in that the reagent (or reaction solution) is not mixed with the sample promptly as a space for sufficiently and uniformly mixing the sample with the reagent is not provided. Moreover, a problem occurs in that a mixing mechanism is not formed easily (for this reason, the reaction channel or the fine channel comprised of a capillary tube is required) as a flow of the sample or reagent (or reaction solution) and a flow of the mixing solution are formed in the same direction. Technical Solution

[8] Accordingly, the present invention has been made in an effort to solve the problems occurring in the related art, and an object of the present invention is to provide a lab- on-a-chip comprising a mixing chamber, in which a sample is mixed with a reagent uniformly and promptly without a capillary tube which is difficult to be processed, density of the sample can be measured accurately by measuring light emitted when the reagent is reacted with the sample, and reproduction can be achieved certainly.

[9] In order to achieve the above object, according to one aspect of the present invention, there is provided a lab-on-a-chip comprising a mixing chamber comprises a first panel and a second panel, which are formed of a transparent material, the first panel including: a reagent injection part formed to inject a reagent; a sample injection part formed to inject a sample; a reagent channel extended from the reagent injection part at a predetermined width; a sample injection channel extended from the sample injection part at a predetermined width; a mixing chamber formed at a place where the reagent channel meets the sample channel, mixing a reagent with a sample; a mixing channel extended from the mixing chamber at a predetermined angle with respect to the reagent channel and the sample channel; and a detection part formed at the end of the mixing channel, and the second panel including: a reagent injection hole which is a through hole connected to the reagent injection part; a sample injection hole which is a through hole connected to the sample injection part; and an exhaust hole which is a through hole connected to the detection part, wherein the mixing chamber is wider than the reagent channel or the sample channel.

[10] An angle between the reagent channel and the sample channel is smaller than 180°, the mixing channel is formed between the reagent channel and the sample channel, and an angle between the mixing channel and the reagent channel and an angle between the mixing channel and the sample channel are smaller than 90°.

[11] A width of the mixing channel or the mixing chamber is greater than that of the reagent channel or the sample channel.

[12] The width of the mixing channel or the mixing chamber is equal to or greater than a value obtained by adding the width of the reagent channel to the width of the sample channel.

[13] The width of the mixing channel or the mixing chamber is formed in the range of

300 to 900μm, and the width of the reagent channel and the sample channel is formed in the range of 150 to 450μm.

[14] The reagent and the sample flow into the reagent channel and the sample channel in the range of 200 to 760 /i6 per minute.

[15] A flow speed of the reagent and the sample, which flow into the the reagent channel and the sample channel, is in the range of 0.15 to 1.7m/s.

Advantageous Effects

[16] As described above, the lab-on-a-chip comprising a mixing chamber according to the present invention has the following advantages. The mixing chamber 19 is formed to be wider than the reagent channel 15 or the sample channel 17 and mixes the reagent with the sample therein, wherein the reagent and the sample flow into the mixing chamber 19 through the reagent channel 15 and the sample channel 17. In this case, the sample is mixed with the reagent uniformly and promptly without a capillary tube which is difficult to be processed. Also, the angle between the reagent channel 15 and the sample channel 17 is smaller than 180°and the angle between the mixing channel 21 and the reagent channel 15 and between the mixing channel 21 and the sample channel 17 is smaller than 90°, the width of the mixing channel 21 can be widened sufficiently, whereby mixture of the reagent and the sample can be made more certainly. Moreover, the density of the sample can be measured more accurately by more accurately measuring light emitted when the reagent is reacted with the sample, and reproduction can be achieved certainly. Brief Description of the Drawings

[17] FIG. 1 is a plane view illustrating a luminescence detection lab-on-a-chip according to the related art.

[18] FIG. 2 is a plane view illustrating another luminescence detection lab-on-a-chip according to the related art.

[19] FIG. 3 is a schematic plane view illustrating a lab-on-a-chip according to the preferred embodiment of the present invention.

[20] FIG. 4 is a schematic plane view illustrating a first panel of a lab-on-a-chip according to the preferred embodiment of the present invention.

[21] FIG. 5 is a schematic view illustrating a use example of a lab-on-a-chip according to the preferred embodiment of the present invention.

[22] FIG. 6 is a photograph illustrating a luminescence emitted on a lab-on-a-chip according to the preferred embodiment of the present invention.

[23] FIG. 7 is a calibration curve to quantify 1-aminopyrene using a lab-on-a-chip with

ODI CL detection according to the preferred embodiment of the present invention.

[24] FIG. 8 illustrates an installation state to confirm reproducibility of a lab-on-a-chip according to the preferred errbodiment of the present invention.

[25] FIG. 9 is a graph illustrating excellent reproducibility of the lab-on-a-chip with ODI

CL detection according to the preferred errbodiment of the present invention.

[26] *Description of Reference Numbers of Main Parts in the Drawings*

[27] 1: lab-on-a chip 10: first panel

[28] 11: reagent injection part 13: sample injection part

[29] 14: detection part 15: reagent channel

[30] 17: sample channel 19: mixing chamber

[31] 21: mixing channel 30: second panel

[32] 31: reagent injection hole 33: sample injection hole

[33] 35: exhaust hole 51 : reagent injection means

[34] 53: sample injection means 55: reagent injection tube

[35] 57: sample injection tube 59: exhaust tube

[36] 60: analysis system 61: signal line

[37] 63: light-detection means 70: light-shielding box

Best Mode for Carrying Out the Invention

[38] Hereinafter, a lab-on-a-chip comprising a mixing chamber according to the preferred errbodiment of the present invention will be described with reference to the accompanying drawings. Wherever possible, the same description as the related art will be omitted, and the same reference numbers will be used throughout the drawings to refer to the same or like parts.

[39] FIG. 3 is a schematic plane view illustrating a lab-on-a-chip 1 comprising a mixing chamber according to the preferred errbodiment of the present invention, FIG. 4 is a schematic plane view illustrating a first panel 10 of a lab-on-a-chip 1 comprising a mixing chamber according to the preferred embodiment of the present invention, and FIG. 5 is a schematic view illustrating a use example of a lab-on-a-chip 1 according to the preferred errbodiment of the present invention.

[40] As shown in FIGs. 3 and 4, the lab-on-a-chip 1 according to the preferred embodiment of the present invention includes a second panel 30 and a first panel 10, which are made of a transparent material. The first panel 10 includes a reagent injection part 11 formed to inject a reagent reacted with a sample to emit light, a reagent channel 15 extended from the reagent injection part 11 at a predetermined width, a sample injection part 13 formed to inject the sample, and a sample channel 17 extended from the sample injection part 13 at a predetermined width and formed at a predetermined angle with respect to the reagent channel 13. The first panel 10 further includes a mixing chanter 19 formed at a place where the reagent channel 15 meets the sample channel 17. The mixing chamber 19 can have various plane shapes, and according to the preferred embodiment of the present invention, the mixing chamber 19 has a pentagonal shape as shown in FIG. 4.

[41] An angle between the reagent channel 15 and the sample channel 17 extended to the mixing chamber 19 is smaller than 180°, preferably has about 120°. The mixing chamber 19 is downwardly extended from the place where the reagent channel 15 meets the sample channel 17 as shown in FIG. 4. Preferably, a width Ll of the mixing chamber 19 is greater than a width L2 of the reagent channel 15 or the sample channel 17. Preferably, the width Ll of the mixing chamber 19 is greater than a value obtained by adding the width L2 of the reagent channel 15 to the width L2 of the sample channel 17.

[42] By forming the width Ll of the mixing chamber 19 more greatly as above, it is possible to provide a space where the reagent and the sample, which are injected through the reagent channel 15 and the sample channel 17 in both directions, can sufficiently be mixed with each other.

[43] The first panel 10 further includes a mixing channel 21 formed in an opposite direction of the mixing chamber 19, i.e., formed between the reagent channel 15 and the sample channel 17. Preferably, a width of the mixing channel 21 is greater than the width L2 of the reagent channel 15 or the sample channel 17. Preferably, the width of the mixing channel 21 is greater than a value obtained by adding the width L2 of the reagent channel 15 to the width L2 of the sample channel 17.

[44] By forming the mixing channel 21 at a sufficient width as above, it is possible to flow a mixing solution of the sample and the reagent mixed in the mixing chamber 19 at a low flow speed through the mixing channel and reach a detection part 43 with a sufficient time so as to sufficiently observe and detect light emitted from the mixing solution.

[45] It is preferable that an angle between the mixing channel 21 and the reagent channel

15 and an angle θ between the mixing channel 21 and the sample channel 17 are greater than 90°, and are preferably formed at about 60°.

[46] By forming the mixing channel 21 at the angle as above, a flow direction of the reagent or the sample, which flow in the reagent channel 15 and the sample channel 17, is formed in opposite to a flow direction of the mixing solution, which flows in the mixing channel 25, so that the sample is effectively mixed with the reagent in the mixing chamber 19. [47] The second panel 30 is provided with a reagent injection hole 31 which is a through hole connected to the reagent injection part 11, a sample injection hole 33 which is a through hole connected to the sample injection part 13, and an exhaust hole 35 which is a through hole connected to the detection part 14.

[48] In the lab-on-a-chip 1 according to the preferred embodiment of the present invention, the first panel 10 and the second panel 30 have been manufactured using a quartz wafer, and the reagent injection part 11, the sample injection part 13, the detection part 14, the reagent channel 15, the sample channel 17, the mixing chamber 19, and the mixing channel 21 have been formed by and a photolithography technique and a wet chemical-etching process.

[49] The second panel 30 and the first panel 10 are bonded to each other up and down, so that the reagent channel 15, the sample channel 17, and the mixing channel 21 are formed as a flow path between the second panel 30 and the first panel 10, and the mixing chamber 19 is formed as a space where the reagent is mixed with the sample.

[50] The use example of the lab-on-a-chip 1 comprising the mixing chamber which includes the second panel 30 and the first panel 10 will be described with reference to HG. 5.

[51] As shown in FIG. 5, a reagent injection means 51 is connected to the reagent injection hole 31 through a reagent injection tube 55, a sample injection means 53 is connected to the sample injection hole 33 through a sample injection tube 57, and an exhaust tube 59 is connected to the exhaust hole 35, wherein the mixing solution of the reagent and the sample is exhausted to the exhaust tube 59. A light-detection means 63 is provided to be spaced apart from the lower part of the lab-on-a-chip 1 at a predetermined interval, and is connected to a micom which is an analysis system 60, through a signal line 61, whereby photo-detection materials detected from the light-detection means 63 can be stored and analyzed in the micom. The lab-on-a-chip 1 and the light- detection means 63 are provided inside a light- shielding box 70 to certainly detect light emitted from the lab-on-a-chip 1.

[52] An example of the light-detection means 63 includes a photomultiplier tube, and an example of the reagent injection means 51 and the sample injection means 53 includes a syringe pump.

[53] Embodiment 1

[54] A quantitative experiment of 1-Aminopyrene was performed using the lab-on-a-chip

1 comprising a mixing chamber according to the present invention, wherein the lab- on-a-chip 1 is installed as shown in FIG. 5. In order to quantify 1-Aminopyrene, l,l'-oxalydiimidazole (ODI) chemical luminescence method was used (see US Patent No. US7,141,677B2). ODI chemical luminescence material, 1-Aminopyrene, and hydrogen peroxide mixture were injected into two syringes. These materials were injected through the reagent injection hole 31 and the sample injection hole 33 of the lab-on-a-chip 1 at a speed of about 200 /i6 per minute by using a syringe pump. The above materials were flowed to the mixing chamber 19 through the reagent channel 15 and the sample channel 17, were promptly mixed with one another in the mixing chamber 19, and then reached the detection part 14 through the mixing channel 21. The mixture which reached the detection part 14 emitted strong light as shown in FIG. 6. Intensity of this light was measured through the photomultiplier tube, and the measured result was stored and analyzed in the micom 60. In FIG. 5, PEEK tube having an outer diameter of 0.8mm and an inner diameter of 0.25mm was used as the reagent injection tube 55 and the sample injection tube 57.

[55] The following Table illustrates intensity of chemical luminescence measured with respect to various concentrations of 1-Aminopyrene

[56] [57] Table 1 [Table 1] [Table ]

[58] FIG. 7 is a gauging line graph illustrating the result obtained in Table 1. [59] In the above experiment, the width L2 of the reagent channel 15 and the sample channel 17 was in the range of 150 to 450μm and their depth was formed at about 50μm. The width Ll of the mixing chamber 19 and the mixing channel 21 was in the range of 300 to 900μm and their depth was formed at about 50μm. A radius of the detection part was in the range of 1.0mm and its depth was formed at about 50μm, and a length of the mixing channel 21 was in the range of 10mm.

[60] The reagent and the sample were flowed through the reagent channel 15 and the sample channel 17 at a speed of about 200 //# per miniute. As a result of the experiment, in the lab-on-a-chip 1 having the dimension range as above, if the flow speed of the reagent and the sample was greater than 200 /i6 per minute, mixture in the mixing chamber 19 was performed uniformly and promptly. Therefore, in order to mix the reagent with the sample in the mixing chamber 19 promptly and uniformly, the flow speed of the reagent and the sample in the reagent channel 15 and the sample channel 17 is preferably in the range of 0.15 to 1.7m/s, more preferably 0.4 to 0.6m/s.

[61] Embodiment 2

[62] A lab-on-a-chip 30 according to the preferred embodiment of the present invention was installed as shown in FIG. 8, so as to carry out a reproduction experiment of the lab-on-a-chip 30. As a result, the result shown in FIG. 9 was obtained.

[63] In FIG. 8, a reference numeral 52 denotes a solvent injection means, 54a a solvent injection tube, 54b a reagent injection tube, 56 a valve for selectively injecting a solvent and a sample into the lab-on-a-chip 30 through an injection tube, 57a an injection tube into which the sample and the solvent are selectively injected, C a waste water vessel for collecting a waste sample or solvent, and 58 a waste water collecting tube connected between the valve 56 and the waste water vessel C.

[64] In a state that the lab-on-a-chip 30 is installed as shown in FIG. 8, the reagent, the solvent, and the sample are injected from the reagent injection means 51, the solvent injection means 52, and the sample injection means 53 into the lab-on-a-chip 30 at a constant speed, wherein the reagent injection means 51, the solvent injection means 52 and the sample injection means 53 are syringe pumps. At this time, the sample and the solvent are selectively injected into the lab-on-a-chip 30 by the valve 56. When the sample, or the solvent or reagent was injected into the lab-on-a-chip 30, chemical luminescence was not observed before the sample was injected thereinto. Chemical luminescence was promptly observed as shown in FIG. 9 when the sample was injected into the lab-on-a-chip 30.

[65] As noted from FIG. 9, light of the same intensity was detected when the sample of the same concentration was injected into the lab-on-a-chip 30.

[66] Accordingly, when the lab-on-a-chip 30 according to the present invention is used, it is possible to quantify exactly sample concentration, and obtain excellent reproducibility within statistical error range. Detection limit of 1-aminopyrene determined using the lab-on-a-chip 1 with ODI detection according to the present invention was as low as 0.3 nM. Industrial Applicability

[67] It will be apparent to those skilled in the art that the present invention can be embodied in other specific forms without departing from the spirit and essential char- acteristics of the invention. Thus, the above embodiments are to be considered in all respects as illustrative and not restrictive. The scope of the invention should be determined by reasonable interpretation of the appended claims and all change which comes within the equivalent scope of the invention are included in the scope of the invention.

Claims

[1] A lab-on-a-chip 1 comprising a mixing chamber, the lab-on-a-chip comprising a first panel 10 and a second panel 30, which are formed of a transparent material, the first panel 10 including: a reagent injection part 11 formed to inject a reagent; a sample injection part 13 formed to inject a sample; a reagent channel 15 extended from the reagent injection part 11 at a predetermined width; a sample injection channel 17 extended from the sample injection part 13 at a predetermined width; a mixing chamber 19 formed at a place where the reagent channel 15 meets the sample channel 17, mixing a reagent with a sample; a mixing channel 21 extended from the mixing chamber 19 at a predetermined angle with respect to the reagent channel 15 and the sample channel 17, and a detection part 14 formed at the end of the mixing channel 21, and the second panel 30 including: a reagent injection hole 31 which is a through hole connected to the reagent injection part 11; a sample injection hole 33 which is a through hole connected to the sample injection part 13; and an exhaust hole 35 which is a through hole connected to the detection part 14, wherein the mixing chamber 19 is wider than the reagent channel 15 or the sample channel 17.

[2] The lab-on-a-chip 1 comprising a mixing chamber as set forth in claim 1, wherein an angle between the reagent channel 15 and the sample channel 17 is smaller than 180°, the mixing channel 21 is formed between the reagent channel 15 and the sample channel 17, and an angle between the mixing channel 21 and the reagent channel 15 and an angle between the mixing channel 21 and the sample channel 17 are smaller than 90°.

[3] The lab-on-a-chip 1 comprising a mixing chamber as set forth in any one of claim 1 or 2, wherein a width of the mixing channel 21 or the mixing chamber 19 is greater than a width L2 of the reagent channel 15 or the sample channel 17.

[4] The lab-on-a-chip 1 comprising a mixing chamber as set forth in claim 3, wherein the width of the mixing channel 21 or the mixing chamber 19 is equal to or greater than a value obtained by adding the width of the reagent channel 15 to the width of the sample channel 17.

[5] The lab-on-a-chip 1 comprising a mixing chamber as set forth in claim 4, wherein the width Ll of the mixing channel 21 or the mixing chamber 19 is formed in the range of 300 to 900μm, and the width of the reagent channel 15 and the sample channel 17 is formed in the range of 150 to 450μm.

[6] The lab-on-a-chip 1 comprising a mixing chamber as set forth in claim 5, wherein the reagent and the sample flow into the reagent channel 15 and the sample channel 17 in the range of 200 to 760 /i6 per minute.

[7] The lab-on-a-chip 1 comprising a mixing chamber as set forth in claim 5, wherein a flow speed of the reagent and the sample, which flow into the reagent channel 15 and the sample channel 17, in the range of 0.15 to 1.7m/s.