Embodiments described herein relate generally to an analysis chip for detection of particles in a sample liquid.
Recently, in the field of biotechnology and healthcare, attention has been focused on semiconductor micro-analysis chips on which microfluidic elements such as micro flow channels and detection mechanisms are integrated. The analysis chip of this type can detect particles and biopolymers contained in the sample liquid flowing in a micro flow channel by means of measuring an electrical signal change which occurs when the particles in the sample liquid pass through micropores formed in the flow channel.
In general, according to one embodiment, there is provided an analysis chip for detection of particles in a sample liquid, comprising: a substrate; a channel provided on a surface portion of the substrate; a liquid storage portion provided on a part of the channel to store the sample liquid, holes being provided at a bottom portion of the liquid storage portion to connect the liquid storage portion and the channel; and first electrodes provided in the channel or the liquid storage portion.
A semiconductor micro-analysis chip of the embodiments will be explained hereinafter with reference to the accompanying drawings.
The semiconductor micro-analysis chip of the present embodiment comprises a first microchannel 20 provided on a surface portion of a substrate 10, an insulating film 31 provided on the substrate 10 to cover an upper surface of the channel 20, micropores (holes) 50 provided in the insulating film 31 on the same end side of the channel 20, and detection electrodes (first electrodes) 60 provided on a bottom portion of the channel 20 on the same end side of the channel 20. The channel 20 is a groove shaped channel laying in the X direction of the surface portion of the substrate 10. A bank 32 formed of an insulating film is provided on the insulating film 31 on an end side of the channel 20 to surround the micropores 50, and a liquid storage portion 40 is thereby formed.
In addition, a liquid introduction reservoir 21 is provided on the other end side of the channel 20. In other words, the insulating film 31 is opened on the other end side of the channel 20 and a bank 33 formed of an insulating film is provided to surround the opened portion.
The end side of the channel 20 and the liquid storage portion 40 are adjacent to each other with the insulating film 31 interposed there between. The micropores 50 are provided in the insulating film 31 as particle detecting portions, and the channel 20 and the liquid storage portion 40 are spatially connected via the micropores 50. The micropores 50 are provided at regular intervals in the X direction and the Y direction. The detection electrodes 60 are provided on the bottom surface of the channel 20 to face to the respective micropores 50.
A diameter of each micropore 50 is desirably larger than detected particles. From the viewpoint of detection accuracy, the diameter of the micropore 50 is desirably, slightly larger than the size of the particles to be detected.
The substrate 10 is obtained by forming an insulating film 12 on a Si substrate 11 and further forming an insulating film 13 on the insulating film 12, and the channel 20 is produced by subjecting the insulating film 13 to selective etching and forming a groove. Then, the insulating film 31 of SiO2 or the like is provided on the insulating film 13 to cover the channel 20. Amplifiers 14 and their contact electrodes 15 are provided on the Si substrate 11. In addition, through electrodes 16 penetrating the insulating film 12 are provided to connect with the contact electrodes 15. The detection electrodes 60 are connected to the respective contact electrodes 15 of the amplifiers 14 via the through electrodes 16.
In this structure, the channel 20 is filled with an electrolyte 301 and then a sample liquid 302 is introduced into the liquid storage portion 40 as shown in
In this situation, a GND electrode (second electrode) 70 is set to be in contact with the sample liquid 302 in the liquid storage portion 40. As regards the GND electrode 70, an electrode rod may be inserted from an upper part of the liquid storage portion 40 or an electrode plate may be arranged to be in contact with the sample liquid at an upper part of the liquid storage portion 40 as shown in
In a case where, for example, particles in the sample liquid 302 introduced into the liquid storage portion 40 are negatively charged, the particles contained in the sample liquid 302 are electrophoresed by an electric field generated between the detection electrodes 60 and the GND electrode 70, under the condition that the electric potential of the detection electrodes 60 is set higher than that of the GND electrode 70. Then, the particles move into the channel 20 through the micropores 50. When the particles in the liquid storage portion 40 pass through the micropores 50, the electric resistance at the micropores is increased and the ion current is varied in accordance with the size of the particles. By detecting variation in the ion current, the particles can be detected. The ion current variation detected at the detection electrodes 60 arranged just under the micropores 50 is input to the amplifiers 14 through the through electrodes 16 and the contact electrodes 15. In general, the ion current variation being small, the signals detected at the detection electrode 60 need to be amplified. Arranging the detection electrodes 60 at the bottom of the channel 20 as in the present embodiment, the shortest connection between the detection electrodes 60 and the amplifier 14 can be established via the through electrodes 16 and the contact electrodes 15. That is, it is possible to avoid signal attenuation and the like due to routing of the electrodes and the like. Therefore, the particles can be detected with high accuracy.
In the present embodiment, the particles can be thus detected only by introduction of the sample liquid and the electric observation. For this reason, high-accuracy detection of bacteria, viruses, and the like can easily be implemented. The present embodiment can therefore contribute to technical fields of prevention of spreading of epidemic diseases and food safety by application to simple detection of infectious disease pathogens, food poisoning bacteria, and the like. The present embodiment can also be applied to monitoring harmful substances such as particulate matters in a sample obtained by collecting particles suspended in the air and subjecting the particles to submerged dispersion.
In addition, by arranging a plurality of micropores 50 in the present embodiment, the frequency of passage of the particles through the micropores 50 can be efficiently increased and the detection efficiency can be enhanced. The micropores 50 through which the particles have passed can be specified by providing the detection electrodes 60 corresponding to the respective micropores 50. Furthermore, even if the particles simultaneously pass through different micropores 50, events can be detected separately.
The detection electrodes 60 are drawn to an underlayer through the insulating film 12 forming the bottom surface of the channel 20 and connected to the amplifiers 14 provided just under the insulating film 12. For this reason, the detection signals can be amplified by the amplifiers 14 without increasing noise due to routing of the electrodes, and the like. Inspection can be therefore performed with good accuracy with faint detection signals.
Next, a method of manufacturing the analysis chip of the present embodiment shown in
First, as shown in
Next, as shown in
Next, as shown in
Next, as shown in
Next, the insulating film 13 is formed on the insulating film 12 by CVD or the like to cover the detection electrodes 60. Subsequently with this, as shown in
Next, as shown in
Next, as shown in
Next, as shown in
Finally, as shown in
According to the present embodiment, as described above, the analysis chip can be manufactured in a general semiconductor device manufacturing process using the Si substrate 11. Therefore, in addition to that the analysis chip of this embodiment can detect the particles with high sensitivity micromachining and mass production of the semiconductor technology can be applied to the analysis chip. For this reason, the analysis chip can be manufactured in a very small size, at low costs.
The detection electrodes 60 are provided for the respective micropores 50 in the present embodiment but one detection electrode 60 may be provided for the micropores 50. As shown in
In addition, the liquid introduced into the channel 20 is not limited to the electrolyte but the channel 20 may be filled with the sample liquid.
The present embodiment is different from the first embodiment with respect to a feature that a liquid discharge reservoir 22 is provided in a microchannel 20. In other words, the liquid discharge reservoir 22 is produced by opening the insulating film 31 on one of end sides of the channel 20 and providing a bank 34 so as to surround the opening portion. A liquid introduction reservoir 21 is provided on the other end side of the channel 20, similarly to the first embodiment. Furthermore, a liquid storage portion 40 is provided on a central portion of the channel 20.
In such a structure, a sample liquid or an electrolyte can be introduced from the liquid introduction reservoir 21 and discharged from the liquid discharge reservoir 22, and a smooth flow of the sample liquid or the electrolyte in the channel 20 can be implemented. A risk of taking in air bubbles through micropores 50 when the sample liquid is dropped into the liquid storage portion 40 can be reduced. In addition, if particles moving from the liquid storage portion 40 into the channel 20 through the micropores 50 are retained in the channel 20, the particles may become a cause of noise in an ion current. However, the particles can be discharged efficiently by implementing a smooth flow of the electrolyte in the channel 20 by the above-described structure of the present embodiment. In other words, high-accuracy measurement reducing noise can be performed. Therefore, according to the present embodiment, in addition that the same advantages as those of the first embodiment can be naturally obtained, the reliability can be increased and the accuracy can be made higher by a smooth flow of the electrolyte in the channel 20.
In the present embodiment, banks 33 and 34 for the respective reservoirs 21 and 22, and a bank 32 for the liquid storage portion 40 are formed separately, but may be formed simultaneously. As shown in
The semiconductor micro-analysis chip of the present embodiment comprises a first microchannel (first channel) 20 provided on a surface portion of a substrate 10, an insulating film 31 covering an upper surface of the channel 20, a second microchannel (second channel) 80 provided on the insulating film 31 so as to make overhead crossing with the channel 20, micropores 50 provided in the insulating film 31 at the portion of the overhead crossing of the channels 20 and 80, detection electrodes (first electrodes) 60 provided at the bottom of the channel 20, and a GND electrode (second electrode) 70 provided on a part of the channel 80.
The channel 20 and the channel 80 make overhead crossing at a central portion of the surface of the substrate 10. The channel 20 is produced by processing the surface portion of the substrate 10 so as to be in a groove shape by selective etching. The channel 80 is formed in tunnel shape obtained by surrounding a space which is to be a channel by an insulating film 85.
The liquid introduction reservoir 21 to introduce the sample liquid or the electrolyte is provided on one of end sides of the channel 20, and the liquid discharge reservoir 22 to discharge the sample liquid or the electrolyte is provided on the other end side of the channel 20. The reservoirs 21 and 22 are produced by opening the insulating film 31 on one end side and the other end side of the channel 20 and providing banks 36 and 37 so as to surround the opened portions.
A liquid introduction reservoir 81 to introduce the sample liquid or the electrolyte is provided on one of end sides of the channel 80, and a liquid discharge reservoir 82 to discharge the sample liquid or the electrolyte is provided on the other end side of the channel 80. The liquid introduction reservoir 81 is produced by providing the bank 36 so as to surround a space connecting to one of ends of the channel 80. The liquid discharge reservoir 82 is produced by providing the bank 37 so as to surround a space connecting to the other end of the channel 80. In other words, the bank 36 is common to the liquid introduction reservoirs 21 and 81 and the bank 37 is common to the liquid discharge reservoirs 22 and 82. In addition, the GND electrode 70 is provided on the liquid introduction reservoir 81.
As shown in
Amplifiers 14 and their contact electrodes 15 are provided at corresponding positions just under the detection electrodes 60, in the Si substrate 11. In addition, through electrodes 16 penetrating the insulating film 12 are provided to be connected to the contact electrodes 15, and the through electrodes 16 are connected to the detection electrodes 60.
In this structure, when the sample liquid containing the particles dispersed is introduced from the liquid introduction reservoir 81 of the channel 80, the sample liquid flows in the channel 80. The channel 20 is preliminarily filled with the electrolyte in advance. The liquid in the channel 80 and the liquid in the channel 20 thereby contact via the micropores 50. The liquid introduced into the channel 20 is not limited to the electrolyte but the channel 20 may be filled with the sample liquid.
When a potential difference is made between the detection electrodes 60 and the GND electrode 70 in this state, an ion current flows through the micropores 50. In addition, setting the electric potential of the detection electrodes 60 to be higher than the electric potential of the GND electrode 70, the particles in the sample liquid introduced into the liquid introduction portion 81 are electrophoresed to move into the channel 20 through the micropores 50 by an electric field generated between the detection electrodes 60 and the GND electrode 70 in a case where the particles are negatively charged. When the particles flowing in the channel 80 pass through the micropores 50, the ion current is varied in accordance with the size of the particles. By detecting the ion current variation, the particles can be detected. The ion current variation is input from the detection electrodes 60 arranged just under the micropores 50 to the amplifiers 14 through the through electrodes 16 and the contact electrodes 15. The particles can be therefore detected with high accuracy by amplifying the variation in the ion current value by the amplifiers 14.
In the case that the particles in the sample liquid are positively charged, the sample liquid may be introduced into the channel 20, the electrolyte may be introduced into the channel 80, and the electric potential of the detection electrodes 60 may be set to be higher than the electric potential of the GND electrode 70. In this case, the particles in the sample liquid move from the channel 20 to the channel 80 through the micropores 50. When the particles flowing in the channel 20 pass through the micropores 50, the ion current is varied in accordance with the size of the particles. Alternatively, in the case that the particles in the sample liquid are positively charged, the sample liquid may be introduced into the channel 80, the electrolyte may be introduced into the channel 20, and the electric potential of the detection electrodes 60 may be set to be lower than the electric potential of the GND electrode 70. Thus, in the structure of the present embodiment, the positive or negative charge of the particles and the electric potentials of the detection electrodes 60 and the GND electrode 70 can be combined freely in accordance with the purposes.
In the present embodiment, the particles can be thus detected by introduction of the sample liquid and the electric observation alone. In addition, the detection efficiency can be enhanced by arranging a plurality of micropores 50, because the frequency of passage of the particles through the micropores 50 can be efficiently increased. The same advantages as those of the first embodiment can be therefore obtained.
In addition, the present embodiment also has an advantage that the intersecting portion of the channels 20 and 80 can be smoothly filled with the sample liquid and the electrolyte since two channels 20 and 80 are used and the liquid is allowed to flow in each of the channels.
Next, as shown in
Next, as shown in
According to the present embodiment, as described above, the analysis chip can be manufactured in a general semiconductor device manufacturing process using the Si substrate 11. Therefore, in addition to that the analysis chip of this embodiment can detect the particles with high sensitivity, micromachining and mass production of the semiconductor technology can be applied to the analysis chip. The same advantages as those of the first embodiment can be therefore obtained.
The GND electrode 70 does not need to be formed on the liquid introduction reservoir 81 but may be formed on the liquid discharge reservoir 82. The GND electrode 70 may be provided at a position in contact with the sample liquid or the electrolyte in the channel 80. As shown in the cross-sectional view of
In addition, as shown in
Furthermore, a particle trap mechanism formed of micropillars 26 may be provided on a downstream side of the channel 20. The micropillars 26 are aligned at intervals slightly smaller than a diameter of the particles to be detected.
The division walls 25 are produced by leaving the insulating film 13 in a plate shape with a line-shaped mask when the insulating film 13 is process in a groove shape. Moreover, the micropillars 26 are produced by leaving the insulating film 13 in a pillar shape with a circular mask when the insulating film 13 is process in a groove shape.
The basic structure of the present embodiment is the same as that of the third embodiment, in the viewpoint that the channels 20 and 80 make overhead crossing. The present embodiment is different from the third embodiment with respect to a feature that microchannels 20 and 80 are produced by bonding two substrates 100 and 200 to each other.
The first microchannel 20 is provided on a surface portion of the first substrate 100. The first substrate 100 is substantially the same as the substrate 10 of the first embodiment. More specifically, insulating films 12 and 13, amplifier 14, contact electrodes 15, through electrodes 16, detection electrodes 60 and the like are formed on a Si substrate 11.
The material of the second substrate 200 is, for example, plastic or quartz, and the microchannel 80 is provided by forming a groove on its lower surface. Furthermore, openings for formation of reservoirs are provided in the second substrate 200. Two channels 20 and 80 make overhead crossing by bonding the substrates 100 and 200 interposing an insulating film 31.
If a Si substrate is used as a second substrate 200′, the channel surface is desirably subjected to thermal oxidation and an oxidized film 201 is formed as shown in a cross-sectional view of
Next, as shown in
Next, as shown in
On the other hand, as shown in
Then, by bonding the substrates 100 and 200 interposing the insulating film 31 as shown in
In the present embodiment, the insulating film 31 is provided on the first substrate 100 side before bonding the substrates 100 and 200 but the insulating film 31 may be provided on the second substrate 200 side. In addition, the sacrificial layer 18 may be removed after bonding the substrates 100 and 200 via the holes 86 to 89.
The final structure of the present embodiment is substantially the same as that of the third embodiment and the same advantages as those of the third embodiment can be therefore obtained. In addition to this, the manufacturing process can be simplified and the manufacturing costs can be reduced since the present embodiment can be implemented by bonding the substrates 100 and 200 to each other.
The invention is not limited to the above-described embodiments. The first channel and the second channel in the third and fourth embodiments do not need to intersect but may be partially adjacent to each other as shown in a plan view of
In this structure, too, the particles can be detected by introduction of the sample liquid and the electric observation alone and, furthermore, the frequency of passage of the fine particles through the micropores 50 can be efficiently increased by arranging a plurality of micropores 50. The same advantages as those of the third embodiment can be therefore obtained.
In addition, both the channels 20 and 80 may be channels in the groove shape as shown in a plan view of
The micropores 50 may be shape in a circle or may be formed in a slit shape at the adjacent portion of the channels 20 and 80. Furthermore, the detection electrodes 60 are formed on a side wall of the channel 20 so as to be opposed to the micropores 50, but may be formed on the bottom surface of the channel 20.
In this structure, too, the particles can be detected by introduction of the sample liquid and the electric observation alone and, furthermore, the frequency of passage of the fine particles through the micropores 50 can be efficiently increased by arranging a plurality of micropores 50. The same advantages as those of the third embodiment can be therefore obtained.
In addition, the first electrodes are arranged on the first channel side and the second electrode is arranged on the second channel side or the liquid storage side in the embodiments, but these electrodes may be arranged on opposite sides. Moreover, the number of holes and the number of detection electrodes can be arbitrarily changed in accordance with specifications.
While certain embodiments have been described, these embodiments have been presented by way of example only, and are not intended to limit the scope of the inventions. Indeed, the novel embodiments described herein may be embodied in a variety of other forms; furthermore, various omissions, substitutions and changes in the form of the embodiments described herein may be made without departing from the spirit of the inventions. The accompanying claims and their equivalents are intended to cover such forms or modifications as would fall within the scope and spirit of the inventions.
Number | Date | Country | Kind |
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2016-185493 | Sep 2016 | JP | national |
This application is a divisional of application Ser. No. 15/461,891 filed on Mar. 17, 2017 and is based upon and claims the benefit of priority from Japanese Patent Application No. 2016-185493, filed Sep. 23, 2016, the entire contents of which are incorporated herein by reference.
Number | Date | Country | |
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Parent | 15461891 | Mar 2017 | US |
Child | 17113710 | US |