CN112316994B - Integrated detection chip and method for saccharomycetes - Google Patents
Integrated detection chip and method for saccharomycetes Download PDFInfo
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Abstract
The invention relates to an integrated detection chip and a method for saccharomycetes, and the integrated detection chip comprises a light-induced dielectrophoresis micro-fluidic chip, wherein the light-induced dielectrophoresis micro-fluidic chip comprises first ITO glass, a first channel layer and second ITO glass coated with hydrogenated amorphous silicon on the surface, and the first ITO glass is provided with a first through hole for injecting a cell sample, a second through hole for flowing out waste liquid and a third through hole for transferring strains; the graphene transistor detection chip comprises a substrate layer, a graphene electrode, a second channel layer and a cover plate layer, wherein the graphene electrode is arranged on the substrate layer, the second channel layer is fixed on the substrate layer by the cover plate layer, and a fourth through hole and a fifth through hole are formed in the cover plate layer; the third through hole is connected with the fourth through hole through a connecting pipe. The invention can realize the integrated separation and detection of the fungi and can realize the rapid and accurate detection of the fungi.
Description
Technical Field
The invention relates to the technical field of fungus detection, in particular to an integrated detection chip and method for saccharomycetes.
Background
Yeast is a widely distributed unicellular fungus, and is often used in food fermentation industries such as bread, white spirit, beer, fruit juice and the like. The saccharomyces cerevisiae is a common part of human intestinal flora and is planted on the skin, vaginal mucosa, digestive tract, respiratory tract and other parts of a human body. Saccharomyces cerevisiae is generally considered to be a non-pathogenic microorganism and can be used as a probiotic for preventing and treating diseases such as diarrhea caused by antibiotics. However, over the last 20 years, cases of invasive Saccharomyces cerevisiae infection have increased year by year, particularly in patient populations with immune disorders. Saccharomyces cerevisiae is considered a novel opportunistic pathogen that is generally nonpathogenic, but often becomes a pathogenic species when the patient's immunity is low. The current case report shows that the saccharomyces cerevisiae is closely related to diseases such as dysentery, acute dysentery, urogenital system infection, esophagitis, pneumonia and the like caused by antibiotics. Invasive fungal infections often take several days from bacterial invasion to infection, and in the clinic, the isolation, incubation and detection of cells by biochemical methods often takes several days, which easily misses the optimal diagnosis and early prevention of the associated disease.
In addition to the conventional biochemical diagnostic methods, several advanced Lab on a chip-based methods are used for rapid isolation and precise detection of microorganisms for the purpose of rapid diagnosis. For example, high throughput droplet microfluidic systems are used to isolate, culture and screen anaerobes (w.j.watts, Elife,2020,9, e56998), microfluidic integrated biosensor platforms, including microfluidic paper-based analytical devices, surface plasmon resonance and surface acoustic waves, are used for in situ detection of food-borne pathogens (m.puiu, TrAC trend.anal.chem.,2020,125,115831; a.p.craig, annu.rev.food sci.technol.,2013,4, 369-. However, these methods are technically very challenging, mainly because the concentration of the species to be detected in the sample is much lower than that of other species or cells, which affects the detection accuracy. In addition, the dielectrophoresis method by applying a non-uniform electric field to polarized cells is also frequently used for separation and detection of a small number of cells and microorganisms (E.O.Adekanmbi and S.K.Srivastava, Lab Chip,2016,16, 2148-. However, dielectrophoresis requires fixed physical electrodes, and has the defects of special electrode physical structure design and inflexibility in cell operation. Therefore, as an innovative version of dielectrophoresis, light-induced dielectrophoresis (ODEP) achieves the separation and characterization of different cells by a dielectrophoresis method in which virtual electrodes are formed by light patterning. Studies have reported that the isolation and detection of gastric cancer cells using ODEP (y.z.zhang, sci.advances,2020,6, eaba9628.) are not suitable for the detection of microbial cells because of the great difference in the purity of microbial cells and mammalian cell structures.
Disclosure of Invention
Therefore, the technical problem to be solved by the invention is to overcome the problems that the rapid screening of the yeast sample cannot be carried out and the accurate detection cannot be ensured in the prior art, thereby providing the integrated detection chip and the method for the yeast, which can realize the rapid screening of the yeast sample and can ensure the accurate detection.
In order to solve the technical problems, the integrated detection chip for the yeast comprises a light-induced dielectrophoresis micro-fluidic chip, wherein the light-induced dielectrophoresis micro-fluidic chip comprises first ITO glass, a first channel layer and second ITO glass with hydrogenated amorphous silicon plated on the surface, the first ITO glass is connected with the second ITO glass through the first channel layer, a first channel and a second channel connected with the first channel are arranged on the first channel layer, a first through hole for injecting a cell sample, a second through hole for flowing out waste liquid and a third through hole for transferring strains are arranged on the first ITO glass, the first through hole is communicated with an inlet of the first channel, the second through hole is communicated with an outlet of the first channel, and the third through hole is communicated with an outlet of the second channel; the graphene transistor detection chip comprises a substrate layer, a graphene electrode, a second channel layer and a cover plate layer, wherein the graphene electrode is arranged on the substrate layer, the second channel layer is fixed on the substrate layer by the cover plate layer, a fourth through hole and a fifth through hole are arranged on the cover plate layer, a third channel is arranged on the second channel layer, the fourth through hole is communicated with an inlet of the third channel, and the fifth through hole is communicated with an outlet of the third channel; the third through hole is connected with the fourth through hole through a connecting pipe.
In one embodiment of the invention, the direction of extension of the second channel intersects the direction of extension of the first channel.
In one embodiment of the present invention, an angle formed between the extending direction of the second channel and the extending direction of the first channel is an acute angle.
In one embodiment of the invention, the first ITO glass and the second ITO glass are staggered from each other.
In one embodiment of the present invention, the graphene electrode comprises a substrate, an electrode layer, and single-layer graphene, wherein the electrode layer is located between the substrate and the single-layer graphene.
In one embodiment of the invention, the electrode layers are uniformly distributed by pairs of electrodes.
In one embodiment of the present invention, a groove is disposed on the substrate, and the graphene electrode is disposed in the groove.
In an embodiment of the present invention, the third channel is parallel to a length extending direction of the graphene electrode along a length direction, and a central axis of the third channel coincides with a central axis of the substrate layer.
In an embodiment of the present invention, a third port is disposed in the third through hole, a fourth port is disposed in the fourth through hole, and the third port is connected to the fourth port through the connecting pipe.
The invention also provides an integrated detection method of the yeast, which is used for detecting the yeast based on any one of the integrated detection chips of the yeast and comprises the following steps: step S1: arranging a static light pattern array at the crossing position of the light-induced dielectrophoresis microfluidic chip and forming a virtual electrode; step S2: injecting a sample solution at an inlet of the light-induced dielectrophoresis microfluidic chip; step S3: resetting a dynamic light pattern array at the crossing position of the light-induced dielectrophoresis microfluidic chip, and moving the screened yeast cells to the outlet of the second channel; step S4: yeast cells in the light-induced dielectrophoresis micro-fluidic chip are transferred to a third channel of the graphene transistor detection chip through a connecting pipe; step S5: and collecting graphene transistor signals, and analyzing and processing to obtain strain characteristics.
Compared with the prior art, the technical scheme of the invention has the following advantages:
compared with a clinical biochemical detection method, the integrated detection chip and the method for the saccharomycetes can realize integrated separation and detection of the fungi, can realize quick and accurate detection of the fungi, and provide a new scheme for timely diagnosis and early prevention of invasive yeast infection diseases.
Drawings
In order that the present disclosure may be more readily and clearly understood, reference is now made to the following detailed description of the embodiments of the present disclosure taken in conjunction with the accompanying drawings, in which
FIG. 1 is a schematic diagram of an integrated detection chip for yeast of the present invention;
FIG. 2 is a schematic diagram of a light-induced dielectrophoresis microfluidic chip according to the invention;
FIG. 3 is a schematic diagram of a graphene transistor detection chip according to the present invention;
FIG. 4 is a schematic diagram of the rapid strain isolation process of the present invention;
FIG. 5a is a graph showing the result of Saccharomyces cerevisiae separation at low capture rate according to the present invention;
FIG. 5b is a schematic diagram showing the result of Saccharomyces cerevisiae separation at high-speed capture according to the present invention;
FIG. 6a is a schematic diagram of the change of the conductance amplitude of the graphene transistor in the presence and absence of yeast cells, respectively, according to the present invention;
FIG. 6b shows the conductance variation of the graphene transistor caused by Saccharomyces cerevisiae with different concentrations.
The specification reference numbers indicate: 10-light-induced dielectrophoresis microfluidic chip, 11-first ITO glass, 111-first through hole, 112-second through hole, 113-third through hole, 12-first channel layer, 121-first channel, 122-second channel, 13-second ITO glass, 141-first interface, 142-second interface, 143-third interface, 20-graphene transistor detection chip, 21-base layer, 22-graphene electrode, 23-second channel layer, 231-third channel, 24-cover plate layer, 241-fourth through hole, 242-fifth through hole, 251-fourth interface, 252-fifth interface and 30-connecting pipe.
Detailed Description
Example one
As shown in fig. 1, 2 and 3, the present embodiment provides an integrated detection chip for yeast, including: a light-induced dielectrophoresis microfluidic chip 10, wherein the light-induced dielectrophoresis microfluidic chip 10 includes a first ITO (indium tin oxide) glass 11, a first channel layer 12, and a second ITO glass 13 coated with hydrogenated amorphous silicon on the surface, the first ITO glass 11 is connected to the second ITO glass 13 through the first channel layer 12, the first channel layer 12 is provided with a first channel 121 and a second channel 122 connected to the first channel 121, the first ITO glass 11 is provided with a first through hole 111 for injecting a cell sample, a second through hole 112 for discharging a waste liquid, and a third through hole 113 for transferring a strain, the first through hole 111 is communicated with an inlet of the first channel 121, the second through hole 112 is communicated with an outlet of the first channel 121, and the third through hole 113 is communicated with an outlet of the second channel 122; the graphene transistor detection chip 20 comprises a substrate layer 21, a graphene electrode 22, a second channel layer 23 and a cover plate layer 24, wherein the substrate layer 21 is provided with the graphene electrode 22, the cover plate layer 24 fixes the second channel layer 23 on the substrate layer 21, the cover plate layer 24 is provided with a fourth through hole 241 and a fifth through hole 242, the second channel layer 23 is provided with a third channel 231, the fourth through hole 241 is communicated with an inlet of the third channel 231, and the fifth through hole 242 is communicated with an outlet of the third channel 231; the third through hole 113 is connected to the fourth through hole 241 by a connection pipe 30.
This embodiment the integrated detection chip of yeast includes: a light-induced dielectrophoresis microfluidic chip 10, wherein the light-induced dielectrophoresis microfluidic chip 10 includes a first ITO glass 11, a first channel layer 12, and a second ITO glass 13 coated with hydrogenated amorphous silicon on the surface, the first ITO glass 11 is connected to the second ITO glass 13 through the first channel layer 12, the first channel layer 12 is provided with a first channel 121 and a second channel 122 connected to the first channel 121, the first ITO glass 11 is provided with a first through hole 111 for injecting a cell sample, a second through hole 112 for flowing out waste liquid, and a third through hole 113 for transferring a strain, the first through hole 111 is communicated with an inlet of the first channel 121, the second through hole 112 is communicated with an outlet of the first channel 121, the third through hole 113 is communicated with an outlet of the second channel 122, so that the first channel 121 is a main channel, and the second channel 122 is a slave channel, is beneficial to screening the saccharomycetes; the graphene transistor detection chip 20 comprises a substrate layer 21, a graphene electrode 22, a second channel layer 23 and a cover plate layer 24, wherein the substrate layer 21 is provided with the graphene electrode 22, the cover plate layer 24 fixes the second channel layer 23 on the substrate layer 21, the cover plate layer 24 is provided with a fourth through hole 241 and a fifth through hole 242, the second channel layer 23 is provided with a third channel 231, the fourth through hole 241 is communicated with an inlet of the third channel 231, the fifth through hole 242 is communicated with an outlet of the third channel 231, and the graphene transistor detection chip 20 can obviously identify different concentration characteristics of strains; the third through hole 113 is connected to the fourth through hole 241 through a connecting tube 30, so that the photoinduction dielectrophoresis microfluidic chip 10 and the graphene transistor detection chip 20 can be connected to each other, the connection of samples is realized, the rapid screening of yeast samples is guaranteed, and the accurate detection is guaranteed.
The extension direction of the second channel 122 and the extension direction of the first channel 121 are crossed with each other, thereby facilitating the screening of strains.
In order to prevent particles or cells in the liquid in the first channel 121 from directly flowing to the second channel 122, an angle formed between the extending direction of the second channel 122 and the extending direction of the first channel 121 is an acute angle. That is to say: the second channel 122 forms a first straight line from an inlet to an outlet, a second straight line is formed from a position where the second channel and the first channel are intersected to the outlet of the second channel 122, and an included angle between the first straight line and the second straight line is an acute angle, so that particles or cells in liquid can be prevented from directly flowing to the second channel 122. When the included angle formed between the extending direction of the second channel 122 and the extending direction of the first channel 121 is 45 °, not only particles or cells in the liquid can be prevented from directly flowing to the second channel 122, but also the processing and manufacturing are convenient.
The first ITO glass 11 and the second ITO glass 13 are mutually staggered and used for connecting an external alternating voltage. Specifically, the first ITO glass 11 and the second ITO glass 13 are staggered by 2mm, so that an external alternating voltage can be conveniently connected.
The first ITO glass and the second ITO glass are formed by plating a layer of ITO material with the thickness of 1um on the upper surface of transparent glass with the length, width and height of 2.5cm, 2.5cm and 0.1cm respectively. The second ITO glass 13 is plated with hydrogenated amorphous silicon (a-H: Si). And one surface of the second ITO glass 13 plated with hydrogenated amorphous silicon is connected with the first channel layer 12. Specifically, the ITO glass coated with hydrogenated amorphous silicon is formed by coating a layer of a-H: Si with the thickness of 1um on the upper surface of ITO on the ITO glass.
The first channel layer 12 is a channel structure processed by a laser engraving machine using a double-sided adhesive tape. The layer comprises two channels, the first channel 121 being a master channel and the second channel 122 being a slave channel. The main channel is 20mm long and 0.5mm wide, and the auxiliary channel is 12mm long and 0.4mm wide. A segment of the slave channel intersects the master channel at 1/3 and is angled at 45 deg..
The photoinduction dielectrophoresis microfluidic chip 10 is composed of ITO glass plated with a-H: Si, a microchannel layer and ITO glass from bottom to top, wherein the a-H: Si layer faces upwards, the ITO surface of the ITO glass faces downwards, the upper layer and the lower layer are bonded through double-sided adhesive of the microchannel layer, and the thickness of the whole channel layer is about 80 micrometers. And the ITO glass on the upper layer is provided with a through hole of 0.8mm at each of the three ends of the main channel and the auxiliary channel, and the through holes are used for the liquid to enter and exit the micro-channel. The two through holes of the main channel are respectively used for injecting samples and flowing out waste liquid, and the through holes of the auxiliary channel are used for transferring screened strains.
In order to ensure the sealing performance in the liquid flowing process, a first interface 141 is arranged in the first through hole 111, a second interface 142 is arranged in the second through hole 112, and a third interface 143 is arranged in the third through hole 113, so that the cell sample can be injected into the first through hole 111, waste liquid can be discharged from the second through hole 112, and a strain can be transferred to the graphene transistor detection chip 20 through the third through hole for testing.
As shown in fig. 3, a groove is formed on the base layer 21, and the graphene electrode 22 is disposed in the groove. Specifically, the base layer 21 is located at the lowest position, and is made of PMMA material with the length, width and height of 2.5cm, 2.5cm and 0.5cm respectively. In the middle of which a groove of 1cm by 0.5cm by 0.06cm is arranged.
The graphene electrode 22 includes a substrate, an electrode layer, and single-layer graphene, where the electrode layer is located between the substrate and the single-layer graphene. Specifically, the graphene electrode comprises three layers, wherein the bottom layer is a 1cm × 0.5cm × 0.06cm Si/SiO2 substrate, the middle layer is an electrode layer with the thickness of 400nm, and the top layer is single-layer graphene.
The electrode layer is uniformly distributed by a plurality of pairs of electrodes. Specifically, the electrode layer is by 9 pairs of electrodes evenly distributed, and the position width 10um that is opposite for every pair of electrode is 2um apart.
The third channel 231 is parallel to the length extending direction of the graphene electrode 22 along the length direction, so that the channels can penetrate through all the electrode pairs; since the middle of each pair of electrodes is connected by graphene, the solution flows over the graphene, and the central axis of the third channel 231 coincides with the central axis of the substrate layer 21. The second channel layer 23 is made of double-sided adhesive tape, the length and width of the second channel layer are 2.5cm and 2.5cm respectively, and the middle of the second channel layer is formed into a third channel 231 with the length of 2cm multiplied by 0.4mm through a laser engraving machine.
A third interface 143 is arranged in the third through hole 113, a fourth interface 251 is arranged in the fourth through hole 241, and the third interface 143 is connected with the fourth interface 251 through the connecting pipe 30, so that the flowing tightness of the liquid is ensured.
The cover plate layer 24 is also made of PMMA with the thickness of 2.5cm multiplied by 0.1cm, a pair of through holes with the central symmetry and the distance of 2cm are arranged on the central axis, and the aperture is 0.8 mm.
The graphene transistor detection chip 20 is mounted in the following manner: coating a thin layer of silica gel in the groove of the substrate layer 21, and then placing the graphene electrode 22 in the groove to play a role in water prevention and sealing; a second channel layer 23 is adhered on the upper surface of the base layer 21, the length direction of the second channel layer 23 is parallel to the length extending direction of the graphene electrode 22, and the central axis of the second channel layer coincides with the central axis of the base layer; the cover plate layer 24 is adhered to the other surface of the second channel layer 23, the position of the cover plate layer is overlapped with that of the substrate layer 21, the positions of the fourth through hole 241 and the fifth through hole 242 are overlapped with the two ends of the second channel layer 23, and a fifth interface 252 is arranged in the fifth through hole 242; the base layer 21 and the cover layer 24 are lightly pressed to ensure a thickness of about 80um of the intermediate layer.
In the invention, corresponding plastic pipe joints are respectively connected to the through holes on the uppermost layers of two independent chips, and the light-induced dielectrophoresis microfluidic chip 10 is communicated with a fourth through hole 241 of the graphene transistor detection chip 20 from the outlet of the second channel 122, so that the sample communication can be realized, the rapid screening and the accurate detection of a yeast sample can be realized, the rapid diagnosis and the early prevention of invasive fungal infection can be realized, the strain separation purity can reach 95%, and the detection chip can obviously identify different concentration characteristics of strains.
Example two
The embodiment provides an integrated detection method of yeasts, which is based on the integrated detection chip of yeasts of embodiment one and comprises the following steps: step S1: arranging a static light pattern array at the crossing position of the light-induced dielectrophoresis microfluidic chip 10 and forming a virtual electrode; step S2: injecting a sample solution at an inlet of the light-induced dielectrophoresis microfluidic chip 10; step S3: resetting a dynamic light pattern array at the crossing position of the light-induced dielectrophoresis microfluidic chip 10, and moving the screened yeast cells to the outlet of the second channel 122; step S4: the yeast cells in the light-induced dielectrophoresis microfluidic chip 10 are transferred to the third channel 231 of the graphene transistor detection chip 20 through the connecting tube 30; step S5: and collecting graphene transistor signals, and analyzing and processing to obtain strain characteristics.
In the integrated detection method for yeast described in this embodiment, in step S1, a static light pattern array is disposed at a crossing of the light-induced dielectrophoresis microfluidic chip, and a virtual electrode is formed to block yeast cells; in the step S2, a sample solution is injected into the inlet of the light-induced dielectrophoresis microfluidic chip 10, which is beneficial to detection; in step S3, a dynamic light pattern array is newly disposed at the intersection of the light-induced dielectrophoresis microfluidic chip 10, cells are captured by the dielectrophoresis force generated by light, and the screened yeast cells are moved to the outlet of the second channel 122; in the step S4, the yeast cells in the photo-induced dielectrophoresis microfluidic chip 10 are transferred to the third channel 231 of the graphene transistor detection chip 20 through the connecting tube 30, so that different concentration characteristics of the strains can be identified through the graphene transistor detection chip 20, and the separation purity of the strains can be improved; in step S5, the graphene transistor signal is collected, and the characteristics of the strain are obtained by analysis and processing, and since the integrated detection chip for the yeast detects the yeast according to the first embodiment, the rapid screening of the yeast sample can be realized, and the accurate detection can be ensured.
The following is a detailed description of the embodiments:
in one embodiment, a schematic diagram of a rapid seed separation process based on a light-induced dielectrophoresis chip is shown in FIG. 4. The strain separation process comprises 4 steps: 1) fixing a static light pattern array at the intersection of the first channel 121 and the second channel 122, and injecting a cell sample at the entrance of the first channel 121; 2) applying dielectrophoresis force on the light pattern area through light-induced dielectrophoresis to block the saccharomyces cerevisiae cells in the light pattern area; 3) after the sample completely passes through, removing the static light pattern array, and replacing the static light pattern array with a dynamic light pattern array, wherein the light pattern moves repeatedly according to the corresponding arrow in the figure, and the cell is dragged to the outlet of the second channel 122; 4) and collecting the separated cells at an outlet or directly transferring the cells to the graphene transistor detection chip 20 through liquid.
In yet another embodiment, a yeast solution and a polystyrene bead (diameter 1um) solution were mixed in an amount of 1:1000 as a sample solution and injected into the light-induced dielectrophoresis microfluidic chip 10, as shown in fig. 5a and 5b, showing the separation results for saccharomyces cerevisiae at different flow rates and different spot widths. The light spots of the static light pattern array were set to 20um wide by 30um intervals with an alternating voltage set to 20V and a frequency of 30 kHz. The Saccharomyces cerevisiae capture rate was counted at flow rates of 0.5, 1.0, 1.5 and 2uL/min, respectively, as shown in FIG. 5 a. The result shows that higher yeast capture rate can be obtained at low speed, and the capture rate of the saccharomyces cerevisiae reaches 58.8% at the flow rate of 0.5uL/min, while the capture rate of the polystyrene pellets is 0.7%. At this time, the separation purity of saccharomyces cerevisiae could reach 84% at high capture rate, and fig. 5b shows the effect of different spot widths on the capture rate. The pattern array interval was 30um, the alternating voltage was set at 20V, frequency 30kHz, and solution flow rate was 0.5 uL/min. The result shows that when the width of the light spot is 30um (equal to the light spot interval), the saccharomyces cerevisiae can obtain the maximum capture rate which is about 68.9%; and the trapping rate for polystyrene beads was 0.72%. At the moment, the separation purity of the saccharomyces cerevisiae can reach more than 95%.
In another embodiment, the detection function of the graphene transistor detection chip on the saccharomyces cerevisiae characteristics is verified. Figure 6a shows the change in conductance amplitude of a graphene transistor with and without yeast cells, respectively. The results show that although solutions with and without yeast cells cause a change in conductance, the rate of change in conductance with yeast cells is greater. This also verifies that the graphene transistor is capable of detecting yeast cells. FIG. 6b shows the conductance changes of graphene transistors caused by Saccharomyces cerevisiae with different concentrations. The result shows that the graphene transistor can obviously distinguish saccharomyces cerevisiae with different concentrations, and the conductivity change rate is larger when the concentration is lower. The result shows that the graphene transistor detection chip can clearly distinguish the concentration characteristics of saccharomyces cerevisiae, and provides a new method for rapid diagnosis of invasive fungal infection.
The terms first, second and the like in the description and in the claims of the present application are used for distinguishing between similar elements and not necessarily for describing a particular sequential or chronological order. It is to be understood that the data so used is interchangeable under appropriate circumstances such that the embodiments of the application are capable of operation in sequences other than those illustrated or described herein.
It should be understood that the above examples are only for clarity of illustration and are not intended to limit the embodiments. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. And obvious variations or modifications therefrom are within the scope of the invention.
Claims (8)
1. An integrated detection chip for yeasts is characterized by comprising:
the light-induced dielectrophoresis micro-fluidic chip comprises first ITO glass, a first channel layer and second ITO glass with hydrogenated amorphous silicon plated on the surface, wherein the first ITO glass is connected with the second ITO glass through the first channel layer, a first channel and a second channel connected with the first channel are arranged on the first channel layer, a first through hole for injecting a cell sample, a second through hole for flowing out waste liquid and a third through hole for transferring strains are arranged on the first ITO glass, the first through hole is communicated with an inlet of the first channel, the second through hole is communicated with an outlet of the first channel, the third through hole is communicated with an outlet of the second channel, and an included angle formed between the extension direction of the second channel and the extension direction of the first channel is an acute angle;
the graphene transistor detection chip comprises a substrate layer, a graphene electrode, a second channel layer and a cover plate layer, wherein the graphene electrode is arranged on the substrate layer, the second channel layer is fixed on the substrate layer by the cover plate layer, a fourth through hole and a fifth through hole are arranged on the cover plate layer, a third channel is arranged on the second channel layer, the fourth through hole is communicated with an inlet of the third channel, and the fifth through hole is communicated with an outlet of the third channel;
the third through hole is connected with the fourth through hole through a connecting pipe.
2. The integrated yeast detection chip according to claim 1, wherein: the first ITO glass and the second ITO glass are mutually staggered.
3. The integrated yeast detection chip according to claim 1, wherein: the graphene electrode comprises a substrate, an electrode layer and single-layer graphene, wherein the electrode layer is located between the substrate and the single-layer graphene.
4. The integrated yeast detection chip according to claim 3, wherein: the electrode layer is uniformly distributed by a plurality of pairs of electrodes.
5. The integrated yeast detection chip according to claim 3, wherein: the substrate is provided with a groove, and the graphene electrode is arranged in the groove.
6. The integrated yeast detection chip according to claim 1, wherein: the third channel is parallel to the length extending direction of the graphene electrode along the length direction, and the central axis of the third channel is overlapped with the central axis of the substrate layer.
7. The integrated yeast detection chip according to claim 1, wherein: and a third interface is arranged in the third through hole, a fourth interface is arranged in the fourth through hole, and the third interface is connected with the fourth interface through the connecting pipe.
8. An integrated detection method of yeast, which is based on the integrated detection chip of yeast of any one of claims 1 to 7, and is characterized by comprising the following steps:
step S1: arranging a static light pattern array at the crossing position of the light-induced dielectrophoresis microfluidic chip and forming a virtual electrode;
step S2: injecting a sample solution at an inlet of the light-induced dielectrophoresis microfluidic chip;
step S3: resetting a dynamic light pattern array at the crossing position of the light-induced dielectrophoresis microfluidic chip, and moving the screened yeast cells to the outlet of the second channel;
step S4: yeast cells in the light-induced dielectrophoresis micro-fluidic chip are transferred to a third channel of the graphene transistor detection chip through a connecting pipe;
step S5: and collecting graphene transistor signals, and analyzing and processing to obtain strain characteristics.
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