TWI253435B - Loop micro fluid system - Google Patents

Abstract

The present invention provides a loop micro fluid system, and the micro fluid therein could be driven to move around the loop passage repeatedly by the air comes from the air holes and through the driving passages. In order to process the polymerase chain reaction (PCR), the micro fluid should bear the temperature changes for three times. Hence, plural temperature controllers are utilized to adjust the temperatures of the sections constituting the loop passage. Moreover, the times of cycles may be determined based on the desired times of reactions. The present invention could also increase or decrease the number of the air holes, the driving passages and the temperature controllers to adjust the times of temperature changes according to the demands of various biochemical reactions.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention This invention relates to a microfluidic motion system, and more particularly to a loop-type microfluidic motion system utilizing a loop-type microfluidic channel. [Prior Art] The development of science and technology has to rely on various biochemical experiments of human operations in the past. It has been able to be carried out by biochips (b i 〇 c h i p ) with computer manipulation, and even complete micro fluid movements.

Polymerase Chain Reaction (PCR) is a common application for microfluidic movement. It can accurately search for specific bases up to hundreds of bp in lengths of tens of millions of base pairs of nucleic acid molecules. Base sequence, and copy this sequence of sequences more than a million times. The PCR is carried out mainly in three successively repeated steps, starting with a double stranding and raising the temperature to ninety-four degrees Celsius, thereby opening the double-strand structure of the D N A template (t e m p 1 a t e ). Inductive pairing is then performed to reduce the temperature to fifty to sixty-five degrees Celsius, allowing a pair of primers to enter the double stranded DNA molecule and bind to the position of the complementary base on the DNA template. The last step in this cycle is DNA synthesis, which raises the temperature to sixty-five to seventy-five degrees Celsius, activates the polymerase, and uses the two primers as a starting point to synthesize a new double-stranded nucleic acid molecular chain.

DNA 5 is carried out by continuously repeating the above three temperature-changing steps in sequence

1253435 The so-called polymerase chain reaction occurs when the melting, binding and chain extension reactions occur to complete the replication. It takes two to three hours for PCR to be carried out in a conventional PCR machine. In recent years, with the development of microelectromechanical process technology, it has been possible to etch a microchannel on a glass substrate and use a constant temperature copper block as a heat source to form a continuous flow. Biochip. The biochip is not only small in size, but also has a low circulation speed and can be connected to a capillary electrophoresis wafer to form an analysis system. In recent years, research has focused only on changing the heating material of the PCR wafer, the size of the microchannel, the material of the wafer, or the number of cycles. The main design principles are roughly the same, and the single direction is still used. This continuous flow mode must be increased in accordance with the number of reactions or the length of the flow path. Generally, the optimal number of cycles for the PCR reaction is about ten to thirty times, so that not only a longer microchannel is required, but also the number of cycles is arbitrarily selected. In the conventional technique, the microfluidic motion device for PCR uses the direction forward and backward as the direction of movement of the microfluid. For example, the "gas microfluidic drive system and method" disclosed in the Patent No. 4 9 9 3 2 of the Republic of China is to drive the microfluids by gas, and the proper operation of the intake and exhaust gases to continuously advance and retreat the microfluids. Reach the effect of reciprocating motion. For example, the Republic of China Patent No. 5 2 8 8 3 6 "Microfluidic Driving Method and Apparatus" provides a pneumatic fluid reciprocating guiding system and a temperature device for biochemical reactions. If this type of method is only suitable for a simple biochemical reaction, the number of PCRs is used to detect the DN A source material, and its freewheeling is reduced to two. Single Chinese dynamic use and post announcement for control without 6

The 1253435 method deals with complex biochemical reactions or chain reactions. In particular, PCR requires constant changes in the temperature of the microfluid, so that the simple reciprocating motion described above requires a variable temperature control, thereby increasing the degree of experimentation or reaction control. In addition, Giordano et al. also published a microcavity polymerization wafer (BC Giordano, J. Ferrance, S. S w AFR Huhmer, and JP Landers, “Pol) Chain Reaction in Polymeric Microchips Amplification in Less Than 2 4 0 Seconds” Biochemistry 291, pp. 124-132, 200 1), the principle is that the sample is placed in the wafer chamber, and the temperature control system is used to raise and lower the temperature to reach the three-temperature cycle, so although it can be arbitrarily determined, it must still be In order to accurately and quickly carry out the temperature recovery, the prior art also provides a microfluidic thermal cycle fluid for reciprocating motion in a single direction as well as thermal cycling. Although the length of the tunnel is reduced and the number of reactions can be arbitrarily controlled, temperature control is still required. ο ο pp et al. continuous flow PCR made of glass substrate using micro-pump unidirectional driving fluid, which continuously flows through the reaction zone of three degrees and fixed for twenty cycles (Kopp, deMello, AJ; Manz, A., Science 1998 pp. 1 Ob eid et al. also use one-way continuous flow to make PCR. There are still only a few fixed cycles to choose from. (If you use the extremely complex enzyme chain edberg, Μη Erase : DN A , Anal will control the reaction quickly and inversely. The system will reduce the flow to change the wafer ^ fixed temperature MU; 04 6 ) ° wafer 5 Ο beid, 7 1253435

Pierre J . ; Christopoulos, Theodore K.

Continuous-flow DNA and RNA amplification chip combined with laser-induced fluorescence detection" A na 1 ytica Chimica Acta Volume: 494, Issue: 1-2, October 8, (2003), pp. 1-9). SchneegaB et al. PCR wafers were also produced in a unidirectional continuous flow mode and provided only thirty-two reaction cycles (Schneega B, Ivonne; Kohler, Johann Michael, "Flow-through polymerase chain reactions in chip thermocyclers M , Molecular Biotechnology Volume: 82, Issue 2, December, 2001, pp. 101-121) ° In summary, the conventional microfluidic driving and motion devices are mainly based on single-direction motion, or the micro-fluid is continuously moving forward and backward. In the case of current PCR biochips, only one-way flow is still used, so a longer flow path is required. The longer the flow path, the higher the frequency of failure, and only a fixed number of reaction cycles can be performed. Micro-chamber Although the PCR wafer and the reciprocating fluid temperature control device can freely change the number of cycles of the reaction, it is still necessary to perform rapid temperature change control to improve the chain reaction. In view of the above problems, the present invention provides a loop-type microfluidic motion system and apparatus for repetitively moving a microfluid in a loop-type microfluidic channel therein, wherein temperature control is utilized. The device adjusts the temperature of each segment of the loop-type microfluidic channel to make the microfluid every 8

1253435 A three-temperature cycle can be completed by a circular motion, and the microfluidic (sample) is optimally reacted by the number of times the coil is moved to obtain an accurate reaction result. SUMMARY OF THE INVENTION It is an object of the present invention to provide a loop-type microfluidic motion system that includes a loop-type microfluidic channel for containing microfluidic motion therein and for gas to enter and exit a plurality of pores. In addition, a plurality of driving microchannels are provided for the microfluidics or gas passages. Each of the driving microchannels is connected to one of the above-mentioned returning microfluidic channels, and the other end is connected to one of the pores. A body supply device is coupled to the plurality of vents for ingress of gas into or out of the vent, thereby driving the microfluidics within the looped microfluidic channel. One less temperature control device is coupled to the looped microfluidic channel to control the temperature of the microfluidic fluid flowing through the looped microfluidic channel. Another object of the present invention is to provide a loop-type microfluidic device comprising a loop-type microfluidic channel for containing microfluidic movement therein and a plurality of pores for gas or fluid to enter and exit. In addition, a plurality of driving microchannels are provided for the passage of fluid or gas, wherein one end of each channel is connected to the loop-type microfluidic channel and the other end is connected to one of the gas channels L. [Embodiment] The present invention will be described in conjunction with the preferred embodiments and the accompanying drawings. It should be understood that all of the preferred embodiments of the present invention are only for the control of the present invention. The body is slightly above the display 9

1253435 is used, so the present invention is applied to other embodiments, except for the preferred embodiment. The present invention is not limited to the embodiment, and the structure of the loop wafer of the preferred embodiment of the present invention is presented in the accompanying patent application and its equivalent. 4) Referring to Figure 1, the loop-type microfluidic channel (10) 1, the main body, and contains several driving micro-flow channels (4, 5, 6) and 2, 3). It should be understood that although only three flow paths and three corresponding air holes are shown in FIG. 1, it is only used to illustrate the preferred embodiment, that is, the application requires three temperature differences, so the present invention can be applied to other implementations. For example, the temperature difference is also in the three stages, and it is necessary to increase or decrease the number of the air holes, in which the number of the air holes and the driving micro flow path, etc., correspond to each other. In addition, the present invention is mainly applied to a sample in the form of a bulk, and therefore the diameter of the micro-circular microfluidic channel driven in the present invention is preferably one hundred micrometers. The description of the embodiment is mainly for the present invention. In PCR), the application of the invention is not limited to PCR. Since one: the sample needs to be repeatedly subjected to the three-temperature cycle, the loop is slightly differentiated into three segments 10 to 12 by the externally driven microchannels 4 to 6. And use three temperatures to 9 to individually control the temperature of each segment, because the body; the body channel is a loop design, so that the microfluid can move in the loop (circumference), and continuously pass three temperatures in sequence > can also be broadly based on any real field. Microchannels, 12) are stomata (1, strip drive micro-example PCR, not only for external channels and meshes with microfluidic flow channels and right. Enzyme linkage, vice versa, & PCR In the example, the fluid channel controller 7 defines the microflow in which the MAN section 10 10 1253435 to 12 is performed, and the effect of the three-temperature cycle is easily obtained. The temperature controllers 7 to 9 can be various heat sources such as metal contacts. Heating, infrared heating, etc., and by the loop design of the present invention, the temperature controllers 7 to 9 can be maintained at a fixed temperature, thereby achieving the three-stage temperature difference without requiring a high degree of difficulty. Variable temperature control. Furthermore, since the inspection system in the form of microfluidics performs a circular motion in the above-mentioned loop-type microfluidic channel, that is, clockwise or reverse

Movement in the direction of the needle, so the number of reactions can be controlled by adjusting the number of turns, whether it is tens or even thousands of reactions, without extending or amplifying the length or volume of the loop microfluidic channel It simplifies the structure of the device, not only saves space, but also reduces the probability of device failure. Further, although the loop-type microfluidic channel exemplified in the present invention is circular, it is not limited thereto, and the present invention can also utilize any microfluidic channel forming a closed loop, such as an elliptical shape or even a multi-turned corner. Fig. 2 is a perspective schematic view showing a loop-type microchannel wafer (loop type microfluidic motion device) of the present invention, which has a size of about three to four centimeters and a thickness of about one millimeter. As shown in FIG. 2, the loop-type microfluidic channel 2 1 of the present invention (that is, the segments 10 to 12 in FIG. 1 ) and the external channels 4 to 6 are all located in the substrate 20 and have the pores 1 to 3 As its entrance and exit with the outside it, it can allow microfluidic (sample) or pressurized gas to enter and exit. In one embodiment of the present invention, the wafer is formed by etching all of the 111535343 tracks (including 1 to 6 and 21) on the underlying substrate and covering it with an upper substrate or other material. It should be understood that the manner in which the loop microfluidic wafer is fabricated is not the focus of the present invention, and thus the present invention should be able to fabricate the wafer using a variety of existing techniques, and is not limited to any particular embodiment. . Since the microfluidic wafer is designed in a loop-like configuration, it will be able to maintain such a small size, regardless of the biochemical reaction within it, which will undergo multiple multiple temperature cycles. Figure 3 presents a microfluidic motion control system architecture diagram that uses a gas to drive the microfluidics within the wafer for loop motion, either clockwise or counterclockwise. The gas source is a gas supply device 32, which is composed of a gas compressor 320, a buffer tank 321 and a plurality of control valves 3 2 2 to 3 2 4, and the number of control valves is at least the number of pores in the wafer 20 Similarly, the three control valves 3 2 2 to 3 2 4 presented in the preferred embodiment of FIG. 3 are connected to the air holes 1 to 3 of the wafer 20, respectively.

The gas compressor 320 further pressurizes the gas to the air pressure required to drive the microfluid, and the pressurized gas is stored in the buffer tank 321, and the buffer tank 3 2 1 is then pressurized. Gas is supplied to the respective control valves 3 2 2 to 3 2 4 . In addition to allowing the pressurized gas to enter the loop-type microfluidic channel via the vent and drive microchannels, the control valve can also vent gas or microfluids in the looped microfluidic channel. The system of Figure 3 can be controlled by the computer 30, and the computer 30 is connected or installed with a data capture card 3 1 . Data capture card 3 1 can transmit 12

The 1253435 drive signal to the gas supply unit 32 to drive fluid movement in the looped microfluidic channel within the wafer 20 by the pressurization of the pressurized gas. When the system is used as a biochemical reaction, 20 is coupled to the heater 34 and the temperature sensor 35, and the heating is transmitted by the computer 30 through the data capture card 31 to provide a heat source. The microfluidic channel (1 〇, 1 1 is heated to various temperatures required for the reaction, and the sensor 35 can detect the temperature in the wafer during the biochemical reaction, and the measured result is transmitted through the data capture card 3 1 It is transmitted to the computer 30 for use as a temperature control. When the biochemical reaction is PCR, the system includes an additional optical detecting device 3, which is directly for detecting the sample in the reaction in the flow channel. Observing the replication efficiency and reaction of the DNA at the time of the observation, this observation is also transmitted to the computer 30 through the data capture card 31, and according to the number of cycles required to determine the reaction, and when the cycle stop body needs to be derived. It should be understood that the configuration of the above-mentioned gas supply device is merely for exemplifying the present invention, and is not intended to limit the use of any gas capable of providing sufficient pressure to enter the air hole and let the gas or microfluid in 20 Further, the microfluidic motion device of the present invention is gas-driven to make the loop type micro; the microfluid in the fluid channel is looped, and the gas is driven by the pores of the wafer 20 The flow path enters the micro-disc device 34, which drives the micro-channels 4 to 6 and the loop-type microfluidic flow into, as shown in Fig. 1, 12) the temperature is measured back to t 3 6 PCR results are determined to be 3 2 The method of the present invention is as follows:

1253435 has an angle of 1 3 to 15 . The angle of this angle will affect the direction of movement of the microfluid in the return microfluidic channel. The configuration in Figure 1 allows the microfluid to be clockwise. However, the present invention is not limited to the configuration of FIG. 1, and the size of the angle can be adjusted to change the direction of motion to counterclockwise motion. The above angle is preferably between zero and forty-five degrees. The angle of the angle is λ!, the driving kinetic energy generated by the gas is larger, but it is also less easy to handle. The excessive angle may cause the injected gas to disperse. The energy of the cycle is insufficient. In addition, the system of the present invention uses a control valve to control the gas or microfluid inlet and outlet, or even closes it. Therefore, the microfluidic channels 4 to 6 of the present invention and the loop-type microfluidic channel have a valveless design to enable the wafer. The internal structure is simplified to save costs and reduce the chance of obstacles. Referring to Fig. 4 and Fig. 5, Fig. 4 shows the driving process of the microfluid in the coil type microfluidic channel of the embodiment, which includes six operating states Α to F, and Fig. 5 shows a flow chart of driving. First, in step 50, the specimen (microfluid) is introduced into the loop-type microfluidic section 46 as shown in state A. According to an embodiment of the present invention, the introduction can be carried out by pushing the sample through-hole 40 and the driving micro-channel 43 into the section 46 by a gas supply means. In step 51, the sample is driven by gas to section 47, as shown in the shape B. During this process, the control valve corresponding to the air hole 40 passes the gas having the appropriate pressure through the driving microchannel 43 into the loop microfluidic channel, and simultaneously closes the control valve corresponding to the air hole 4 1 , and the coil clamp is clamped. The body motion is returned to the road, and the control valve corresponding to the air hole 42 is opened and the air is released from the air hole 42 so that the sample in the section 46 is clockwise. The thrust of the direction is driven to section 47 as shown in state C. Next, in step 52, the sample is driven by gas to section 48. It operates by closing the vent 4 2 so that the pressurized gas enters through the vent 4 1 and opens the vent 40 to allow gas to escape therefrom, as indicated by state D. As a result, a driving force is generated to move the specimen from the segment 47 to the segment 48 as shown in state E.

After the specimen enters section 48, the specimen will be moved to section 46 in step 53 to complete a cycle of clockwise loop motion. It operates by closing the vent 40 so that pressurized gas enters the vent 42 and opens the vent 4 1 to allow gas to escape therefrom, as indicated by state F. Thus, similar to the above, a drive force will be generated to move the specimen from section 48 to section 46, as shown in state A. Since this embodiment is applied to P C R, the respective sections have been adjusted to an appropriate reaction temperature by the temperature control means before the reaction. For example, the temperature in section 46 is raised to 94 degrees Celsius, while section 47 reduces the temperature to fifty degrees, and section 48 again raises the temperature to seventy degrees. Therefore, each round of the loop-type microfluidic channel of the sample can undergo a three-temperature cycle, and through the above repeated operations, the sample can be subjected to the expected number of three-temperature cycles, when the number of cycles is not reached. When it is completed in step 353, it will return to step ί 5 1, and the specimen repeats another clockwise movement. When the desired number of cycles is reached, the sample is exported, as shown in step 5 4, 15

1253435 for precise results. The conventional techniques all use a one-way motion or a reciprocating transporter in a straight line. Although the one-way motion does not require complicated temperature-changing control, the number of times should be limited, and the wafer product may increase and the structure may be complicated due to the increase in the number of times. However, the design of the reciprocating motion can control the number of times and fix the volume, but it requires a relatively difficult temperature control to affect the results of the experiment. The loop type design of the present invention can solve the problems encountered by the above-mentioned prior art, and has the advantages of all the prior art, thereby improving the efficiency of the reaction, and the design of the wide gate flow path reduces the complexity of the wafer process. , increasing the cost advantage of the wafer of the present invention. The present invention has been described by way of example only, and is not intended to limit the scope of the invention. Modifications and similar configurations made without departing from the spirit and scope of the invention are intended to be included in the scope of the appended claims. BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1 is a schematic structural view showing a micro flow path device of a preferred embodiment of the present invention. Figure 2 is a perspective schematic view of a loop-type microfluidic wafer. Figure 3 is a system architecture diagram showing the loop-type microfluidic system of the present invention. Figure 4 shows the driving process of the microfluid in the loop-type microfluidic channel. In order to make the reverse system quick, the time is added. Therefore, the present invention should be similar to the loop body. 16 1253435 Figure 5 shows the flow of the microfluidic driving process. Figure. [Main component symbol description] 1 to 3 Air holes 4 to 6 Drive micro flow path 7 to 9 Temperature control device 1 0 to 12 Loop type microfluidic channel section 1 3 to 1 5 Angle 17

Claims (1)

1253435 X. Patent application scope: A loop-type microfluidic motion system comprising: a substrate having a loop-shaped microfluidic channel, a plurality of pores, and a plurality of driving microchannels, and each of the driving microchannels is connected to the back a ring-shaped microfluidic channel connected to one of the pores, wherein the loop-type microfluidic channel is configured to accommodate movement of microfluids therein for gas or the microfluid to enter and exit, and the plurality a driving microchannel for the microfluid or the gas to pass through; a gas supply device coupled to the plurality of pores for causing the gas to enter or exit the pore to drive the loop micro through the driving microchannel The microfluidic fluid in the fluid channel; and at least one temperature control device coupled to the looped microfluidic channel for controlling the temperature of the microfluidic fluid within the looped microfluidic channel. φ 2. The loop-type microfluidic motion system of claim 1, wherein the loop-type microfluidic channel and the driven microchannel have a diameter of about one hundred micrometers. 3. The loop-type microfluidic motion system of claim 1, wherein the loop-type microfluidic channel has an angle with the driving microchannel for controlling the direction of movement of the microfluid. 4. The loop-type microfluidic motion system as described in claim 3, wherein the direction of motion comprises a clockwise direction and a counterclockwise direction 〇 18 1253435 5 · 6· 8. 9. 1 0. 11. The loop-type microfluidic motion system of claim 3, wherein the angle is between zero and forty-five degrees. The loop-type microfluidic motion system of claim 1, wherein the shape of the loop-type microfluidic channel comprises a circular, elliptical or polygonal shape. The loop-type microfluidic motion system of claim 1, wherein the plurality of gas supply devices are the same as the number of the plurality of pores, and -- corresponding. The loop-type microfluidic motion system of claim 7, wherein at least one of the gas supply devices performs an operation of intake or exhaust through the air hole connected thereto, so that the micro-fluid channel in the loop-type microfluidic channel The fluid moves in one direction. The loop-type microfluidic motion system of claim 8, wherein one of the gas supply devices performs an intake operation and the other of the gas supply devices performs an exhaust operation. The loop-type microfluidic motion system of claim 1, further comprising an optical detecting device coupled to the loop-type microfluidic channel for detecting the microfluid in the loop-type microfluidic channel. The loop-type microfluidic motion system of claim 1, wherein the plurality of temperature controllers respectively control the temperature of each segment in the loop-type microfluidic channel. The loop-type microfluidic motion system of claim 11, wherein at least one of the sections has a temperature different from that of the other sections 19 12. 1253435 1 3. The temperature. A loop-type microfluidic motion device comprising: a substrate having a loop-type microfluidic channel, a plurality of pores, and a plurality of driving microchannels, wherein each of the driving microchannels is connected to the back a ring-shaped microfluidic channel, the other end being connected to one of the air holes; wherein the loop-type microfluidic channel is for accommodating movement of a microfluid for supplying gas or the microfluid in and out, and the plurality Drive microchannels for the microfluid or the gas to pass through. A loop-type microfluidic motion device according to claim 13 wherein the loop-type microfluidic channel and the driven microchannel have a diameter of about one hundred micrometers. The loop-type microfluidic motion device of claim 13, wherein the loop-type microfluidic channel has an angle with the driving microchannel for controlling the microfluid in the loop-type microfluidic channel. The direction of movement within. 16. The loop-type microfluidic motion device of claim 15, wherein the direction of motion comprises a clockwise direction and a counterclockwise direction. 17. The loop-type microfluid as described in claim 15 The motion device, wherein the angle is between zero and forty-five degrees. 18. The loop-type microfluidic motion device of claim 13, wherein the shape of the loop-type microfluidic channel comprises a circle, an ellipse 20 1 9. 2 0. 1253435 Round or polygonal. The loop-type micro-disc according to claim 13 , wherein a plurality of gas supply devices are coupled to the hole for performing an air intake operation through the at least one air hole to make the micro-direction in the loop-type microfluidic channel Perform a loop motion. A loop-type micro-disc as described in claim 19, wherein one of the gas supply and control devices is operated and the other gas supply and control device operates. 2 1. The loop-type micro-disc as described in claim 13 wherein at least one temperature controller is coupled to the body motion device and individually controls the temperature of the loop-type micro-flow section. 22. The loop type micro φ set forth in claim 2, wherein at least one of the sections has a temperature different from the temperature. Fluid movement device The plurality of gas operation or exhaust fluid is supplied to one of the fluid movements to perform the air intake. The exhaust fluid is moved to recirculate the fluid flow in the microfluidic channel.