Biological chip and manufacturing method therefor

Abstract

A biological chip and a manufacturing method therefor. The biological chip comprises a chip body, the chip body is provided with a plurality of groove structures having openings on the same side, and the plurality of groove structures include a first groove and at least one second groove which is closest to the first groove and has a different depth. The depths of the most nearest groove structures in the biological chip are different, so that a mutual interference between optical signals generated by samples in the adjacent groove structures can be reduced, thereby improving the resolution of a fluorescent optical system and increasing the signal-to-noise ratio of the biological chip during detection processes. Furthermore, a processing flux can be improved by increasing the density of the groove structures, and signal quality can be improved by controlling the depths of the groove structures, thereby achieving the compatibility of high flux and high signal quality. In addition, the biological chip allows repeated use, thereby effectively reducing the use cost.

Description

Biological slide and preparation method thereof Technical Field

[0001] The present application relates to the technical field of biological sample detection, and in particular to a biological slide and a preparation method thereof. Background Art

[0002] In the field of biological sample detection technology, microscopic samples (such as DNA samples in gene sequencing) are usually carried on a biological slide (such as a sequencing chip) and then their fluorescence signals are collected. However, existing biological slides have the problem of mutual interference of optical signals.

[0003] Taking sequencing chips as an example, existing sequencing chips generally use chemically modified surfaces (such as surfaces modified with APTMS (3-aminopropyl-trimethoxysilane)) to adsorb biological samples (for example, DNA (deoxyribonucleic acid) molecular nanospheres (DNBs), etc.). Chemically modified patterned array sites are produced on the surface of a silicon substrate through photolithography and chemical vapor deposition (CVD), and then the surface chemical modification (such as APTMS) is used to adsorb biological samples.

[0004] However, samples at adjacent array sites on existing sequencing chips are at risk of interfering with each other, resulting in a reduced signal-to-noise ratio. In addition, due to the scattering and diffraction of optical signals and the limitations of the optical system, it is difficult for existing sequencing chips to continuously improve chip density or throughput without affecting the quality of sequencing signals.

[0005] Summary of the Invention

[0006] In order to solve at least one of the above defects, it is necessary for the present application to propose a biological slide that is conducive to reducing mutual interference between samples on adjacent array sites and improving the signal-to-noise ratio; and can continuously improve the chip density or throughput without affecting the quality of the detection signal.

[0007] In addition, the present application also provides a method for preparing the aforementioned biological slide.

[0008] In a first aspect, an embodiment of the present application provides a biological slide, comprising a slide body having a plurality of groove structures with openings located on the same side, the plurality of groove structures comprising a first groove and at least one second groove that is closest to the first groove and has a different depth.

[0009] In some possible embodiments, a depth difference between the first groove and the second groove closest to the first groove is greater than or equal to 20 nm.

[0010] In some possible embodiments, the plurality of groove structures further include a third groove adjacent to the first groove, the distance between the third groove and the first groove is greater than the distance between the second groove and the first groove, and the depth of the third groove may be the same as that of the first groove.

[0011] In some possible embodiments, the depth of the groove structure is 20 nm to 150 nm.

[0012] In some possible embodiments, the carrier body includes a substrate and at least one adsorption structure layer located on the substrate, the groove structure is formed on the adsorption structure layer, and the first groove and the second groove extend into the adsorption structure layer to different depths.

[0013] In some possible embodiments, the adsorption structure layer includes: an adsorption layer located on the surface of the substrate and an isolation layer located on the surface of the adsorption layer, wherein the adsorption layer is used to adsorb biological samples, the groove structure at least penetrates the isolation layer, and the adsorption layer is exposed by the groove structure.

[0014] In some possible embodiments, all or part of the groove structure extends into the adsorption layer.

[0015] In some possible embodiments, the wafer carrier body includes multiple layers of the adsorption structure layer stacked together, and the first groove and the second groove extend into different layers of the adsorption structure layer.

[0016] In some possible embodiments, the inner diameter of the groove structure is 100 nm to 300 nm; and / or the distance between the centers of two adjacent groove structures is 200 nm to 1000 nm.

[0017] In some possible embodiments, the adsorption layer includes an adsorption material for adsorbing biological samples, the adsorption material includes at least one of metal nitride, metal oxide and metal, and / or the material of the isolation layer includes at least one of metal nitride and metal oxide.

[0018] In some possible embodiments, a buffer layer is further provided between the substrate and the adsorption structure layer, and a material of the buffer layer includes at least one of oxide and nitride.

[0019] In a second aspect, an embodiment of the present application provides a method for preparing a biological slide, the method comprising:

[0020] A plurality of groove structures with openings located on the same side are formed on the slide body of the biological slide, wherein the plurality of groove structures include a first groove and at least one second groove closest to the first groove and having a different depth.

[0021] In some possible embodiments, the step of forming a plurality of groove structures with openings located on the same side on the slide body of the biological slide includes:

[0022] forming the first groove and the second groove on the slide body at the same time; or

[0023] The first groove and the second groove are respectively formed on the carrier body.

[0024] In some possible embodiments, the step of simultaneously forming the first groove and the second groove on the slide body includes:

[0025] forming a patterned film having a first etching hole and a second etching hole on the surface of the carrier body; and

[0026] The portion of the carrier body corresponding to the first etching hole is removed to form the first groove on the carrier body, and the portion of the carrier body corresponding to the second etching hole is removed to form the second groove on the carrier body.

[0027] In some possible embodiments, the step of forming the first groove and the second groove on the slide body includes:

[0028] forming a patterned film having first etching holes on the surface of the carrier body;

[0029] removing a portion of the wafer carrier body corresponding to the first etching hole to form the first groove on the wafer carrier body;

[0030] forming a patterned film having second etching holes on the surface of the carrier body; and

[0031] A portion of the wafer carrier body corresponding to the second etching hole is removed to form the second groove on the wafer carrier body.

[0032] In some possible embodiments, the preparation method further includes:

[0033] A third groove adjacent to the first groove is formed on the carrier body, the distance between the third groove and the first groove is greater than the distance between the second groove and the first groove, and the depth of the third groove and the first groove can be the same.

[0034] In some possible embodiments, the step of forming a plurality of groove structures with openings located on the same side on the slide body of the biological slide includes:

[0035] forming at least one adsorption structure layer on the surface of the substrate; and

[0036] The groove structure is formed on the adsorption structure layer, and the first groove and the second groove extend into the adsorption structure layer to different depths.

[0037] In some possible embodiments, the step of forming at least one adsorption structure layer on the surface of the substrate includes:

[0038] forming an adsorption layer on the surface of the substrate, wherein the adsorption layer is used to adsorb biological samples; and

[0039] forming an isolation layer on the surface of the adsorption layer,

[0040] In the step of forming the groove structure on the adsorption structure layer, the groove structure at least penetrates the isolation layer, so that the adsorption layer is exposed from the groove structure.

[0041] In some possible embodiments, a depth difference between the first groove and the second groove closest to the first groove is greater than or equal to 20 nm.

[0042] In the bioslide and preparation method provided in the embodiments of the present application, the depths of the first and second grooves closest to each other in the groove structure of the bioslide are set to different levels. This groove structure with different depths can greatly reduce the mutual interference of samples between adjacent groove structures, increase the resolution of the fluorescence optical system, and improve the signal-to-noise ratio of the bioslide during the detection process. The throughput of biochemical reactions can be increased by increasing the density of the groove structure, and the quality of optical signals can be improved by controlling the depth of the groove structure. For example, in the field of sequencing, the compatibility of high sequencing throughput and high sequencing signal quality can be achieved. In addition, the bioslide is reusable, effectively reducing the cost of use. BRIEF DESCRIPTION OF THE DRAWINGS

[0043] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the following is a brief introduction to the drawings required for use in the embodiments of the present application. Obviously, the drawings described below are only some embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without any creative work.

[0044] FIG1A is a top view of a biological slide provided in accordance with an embodiment of the present application.

[0045] FIG. 1B is a cross-sectional view along line AA in FIG. 1A .

[0046] FIG2 is a schematic structural diagram of a biological slide provided in another embodiment of the present application.

[0047] 3A and 3B are schematic structural diagrams of a mask 1 and a mask 2 provided in accordance with an embodiment of the present application, respectively.

[0048] 4A to 4D are schematic structural diagrams of mask plates 3 to 6 provided in an embodiment of the present application, respectively.

[0049] FIG5 is a schematic structural diagram of a biological slide provided in yet another embodiment of the present application.

[0050] FIG6 is a schematic structural diagram of a biological slide provided in yet another embodiment of the present application.

[0051] FIG7 is a schematic structural diagram of a biological slide provided in yet another embodiment of the present application.

[0052] FIG8 is a flow chart of a method for preparing a biological slide provided in one embodiment of the present application.

[0053] FIG9 is a flow chart of the simultaneous forming of the first groove and the second groove according to an embodiment of the present application.

[0054] FIG10 is a flow chart showing the step-by-step formation of the first groove and the second groove according to an embodiment of the present application.

[0055] FIG11 is a flow chart of a method for preparing a biological slide provided in another embodiment of the present application.

[0056] FIG12 is a flow chart of a method for preparing a biological slide provided in yet another embodiment of the present application.

[0057] 13A to 13F are schematic diagrams illustrating a process for preparing a biological slide according to an embodiment of the present application.

[0058] 14A to 14F are schematic diagrams of a preparation process of a biological slide provided in another embodiment of the present application.

[0059] 15A to 15F are schematic diagrams of a preparation process of a biological slide provided in yet another embodiment of the present application.

[0060] Description of main component symbols

[0061] The following specific implementation methods will further illustrate the present application in conjunction with the above-mentioned drawings. DETAILED DESCRIPTION

[0062] The following will be combined with the drawings in the embodiments of this application to clearly and completely describe the technical solutions in the embodiments of this application. Obviously, the embodiments described are only part of the embodiments of this application, not all of the embodiments. Based on the embodiments in this application, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of this application.

[0063] It should be noted that when a component is referred to as being "fixed to" or "mounted on" another component, it may be directly on the other component or there may be a central component. When a component is referred to as being "disposed on" another component, it may be directly on the other component or there may be a central component. As used herein, the term "and / or" includes all and any combinations of one or more of the relevant listed items.

[0064] Referring to Figures 1A, 1B, and 2, an embodiment of the present application provides a biological slide 100. The biological slide 100 includes a slide body 10 having multiple slot structures 4 with openings located on the same side. The multiple slot structures 4 include a first slot 4a and at least one second slot 4b that is closest to the first slot 4a and has a different depth. The slot structures 4 can be used to accommodate biological samples, providing a reaction site for the biological samples. That is, each slot structure 4 can serve as a reaction area of ​​the biological slide 100. The biological sample undergoes a biochemical reaction within the slot structure 4, and then the light signal emitted by the product sample can be detected to achieve product detection. Specifically, the biological slide 100 can be used for gene sequencing, for example, as a sequencing chip. It is understood that the biological slide 100 can also be used for other biochemical applications.

[0065] Among them, each first groove 4a in the biological slide 100 has a different depth from the second groove 4b closest to it, so that the biological samples in each first groove 4a and the second groove 4b closest to the first groove 4a are located in different planes. This can reduce the risk of mutual interference (such as interference of fluorescence signals) between samples in different reaction areas closest to each other, which is conducive to improving the signal-to-noise ratio during the detection (such as sequencing) process.

[0066] In some embodiments, the plurality of groove structures 4 further include a third groove 4g adjacent to the first groove 4a. The distance between the third groove 4g and the first groove 4a is greater than the distance between the second groove 4b and the first groove 4a. The third groove 4g and the first groove 4a may have the same depth. As shown in FIG1A , the first groove 4a is surrounded by a plurality of second grooves 4b and a plurality of third grooves 4g arranged in an array. The plurality of second grooves 4b are located approximately on the four sides of the first groove 4a, and the third groove 4g is located approximately diagonally opposite the first groove 4a. The third groove 4g is further away from the first groove 4a than the second groove 4b. In this case, the third groove 4g is less likely to interfere with the optical signal generated by the reaction of the biological sample in the first groove 4a. The depth of the third groove 4g may be the same as that of the first groove 4a. It is understood that the depth of the third groove 4g may also be different from that of the first groove 4a.

[0067] In some embodiments, the depth relationship between two adjacent groove structures 4 can be defined by a preset distance. That is, when the distance between any two adjacent groove structures 4 (i.e., the distance between the centers of two adjacent grooves) is less than or equal to the preset distance, the two adjacent groove structures 4 have different groove depths. When the distance between any two adjacent groove structures 4 is greater than the preset distance, the two adjacent groove structures 4 can have the same or different groove depths. This preset distance is the maximum distance at which optical signal interference occurs between samples within two adjacent groove structures 4. That is, when the distance between two adjacent groove structures 4 is less than or equal to the preset distance, the optical signals within the two groove structures 4 will interfere. When the distance between two adjacent groove structures 4 is greater than the preset distance, the optical signals within the two groove structures 4 will not interfere or the interference will be very weak. As shown in FIG1A , the preset distance can be, for example, the distance L between the adjacent second groove 4b and the first groove 4a. When additional groove structures are formed between the second groove 4b and the first groove 4a to increase the density of the groove structures 4, the groove depth needs to be set different from that of the first groove 4a and the second groove 4b to reduce optical signal interference. It can be seen that the preset distance can be determined according to the actual situation of light signal interference caused by samples in different groove structures 4.

[0068] In addition, by designing different groove structures 4 on the biological slide 100 to have different groove depths, the interference of scattering and diffraction between adjacent fluorescent signals is reduced. In this way, the number of groove structures 4 can be increased by adjusting the groove depth within the range of light signal interference, thereby increasing the density of the groove structures 4 and obtaining higher signal quality. This achieves compatibility between the density of the groove structure 4 and the signal quality, which is beneficial to improving the throughput of the biological slide 100 during the biochemical processing process.

[0069] The bioslide 100 shown in Figures 1A and 1B has groove structures 4 of two depths, wherein the first groove 4a and the third groove 4g are spaced relatively wide apart and may be grooves of the same depth, while the second groove 4b has a different depth than the first groove 4a and the third groove 4g. The bioslide 100 shown in Figure 2 has groove structures of four depths, wherein the spacing between the groove structures 4 decreases and the density of the groove structures 4 increases. When the dimensions of the slide bodies 10 of the bioslides 100 shown in Figures 1A and 2 are substantially identical, adjusting the depths of adjacent groove structures 4 can effectively increase the density of the groove structures 4. Therefore, by designing the groove depths and spacing between the groove structures 4, both the throughput and detection signal quality of the bioslide 100 can be improved. For example, during sequencing, both high sequencing throughput and high sequencing signal quality can be achieved.

[0070] In some embodiments, the depth difference between the first groove 4a and the second groove 4b can be greater than or equal to 20 nm. By controlling the groove depth difference to be greater than 20 nm, the height difference between the light-emitting surfaces of the samples in the closest adjacent reaction areas can be further increased, thereby further reducing the risk of mutual interference and improving the signal-to-noise ratio. It will be appreciated that the specific depth difference between the two closest groove structures 4 can be designed based on actual conditions, for example, the depth difference can be designed based on the required density of the groove structures 4. For example, when the throughput requirement of the biological slide 100 is high, the density of the groove structures 4 can be designed to be larger. In this case, the distance between adjacent groove structures 4 is close, and the degree of optical signal interference is greater. To reduce optical signal interference, the depth difference between adjacent groove structures 4 can be designed to be larger. When the throughput requirement of the biological slide 100 is low, the density of the groove structures 4 can be designed to be smaller. In this case, the distance between adjacent groove structures 4 is far, and the degree of optical signal interference is relatively low. In this case, the depth difference between adjacent groove structures 4 can be designed to be smaller.

[0071] In some embodiments, the depth of the groove structure 4 can be 20nm to 150nm, further 30nm to 100nm. For example, the depth of the groove structure 4 can be 20nm, 30nm, 40nm, 50nm, 60nm, 70nm, 80nm, 90nm, 100nm, 110nm, 120nm, 130nm, 140nm or 150nm. The depth range of the aforementioned groove structure 4 is relatively large. By controlling the groove depths of different groove structures 4 to be within the above range and controlling the difference in the depths of the two closest groove structures 4 to be greater than or equal to 20nm, more groove structures 4 with different depths can be designed, which is conducive to further improving the density of the groove structure 4 and the processing throughput while ensuring high signal quality, thereby achieving compatibility between high throughput and high signal quality.

[0072] The plurality of groove structures 4 can be arranged in an array, ensuring that the nearest groove structures 4 surrounding each groove structure 4 have different depths. The shape of the groove structure 4 and its radial dimensions can be designed based on the required capacity of each reaction area in the biological slide 100. In some embodiments, the groove structure 4 can be a circular groove with an inner diameter of 100 nm to 300 nm, further 120 nm to 250 nm, and further 150 nm to 200 nm. For example, the inner diameter of the groove structure 4 can be 100 nm, 110 nm, 120 nm, 130 nm, 140 nm, 150 nm, 160 nm, 170 nm, 180 nm, 190 nm, 200 nm, 210 nm, 220 nm, 230 nm, 240 nm, 250 nm, 260 nm, 270 nm, 280 nm, 290 nm, or 300 nm.

[0073] In some embodiments, the distance between the centers of two adjacent groove structures 4 (i.e., the groove spacing) can be 200 nm to 1000 nm, further 250 nm to 800 nm, and further 300 nm to 600 nm. For example, the distance between the centers of two adjacent groove structures 4 can be 200 nm, 250 nm, 300 nm, 350 nm, 400 nm, 450 nm, 500 nm, 550 nm, 600 nm, 650 nm, 700 nm, 750 nm, 800 nm, 850 nm, 900 nm, 950 nm, or 1000 nm. By regulating the groove spacing and groove depth, the density of the groove structures 4 can be increased, thereby improving the biochemical processing throughput. Furthermore, because adjacent groove structures 4 have different depths, the bioslide 100 can still have high signal quality even when the groove spacing is small. Therefore, the density of the groove structures 4 can be maximized to produce a higher-throughput bioslide, achieving compatibility between high throughput and high signal quality.

[0074] When forming groove structures 4 with different depths on the wafer carrier 10, a step-by-step molding method can be used. For example, with respect to the structural form of the groove structure 4 in FIG. 1A , where the first groove 4a and the second groove 4b have different depths and the first groove 4a and the third groove 4g have the same depth, two types of masks can be used to achieve this. The first mask shown in FIG. 3A has a first etched hole corresponding to a groove structure 4 of one depth (e.g., the first groove 4a and the third groove 4g), and the second mask shown in FIG. 3B has a second etched hole corresponding to a groove structure 4 of another depth (e.g., the second groove 4b). The groove structures 4 with two different depths as shown in FIG. 1A can be formed by using two masks with different arrangements, the first mask and the second mask, and adopting a step-by-step patterning method. It is understood that in other embodiments, the groove structures 4 with four different depths as shown in FIG. 2 can be formed by using masks with four different arrangements of grooves, such as the third mask to the sixth mask shown in FIG. 4A to FIG. 4D , and adopting a step-by-step patterning method. It is also understood that more types of masks can be used to achieve the formation of groove structures 4 with more different depths, so as to increase the arrangement density of the groove structures 4. It is also understood that the formation of groove structures 4 with different depths is not limited to the above distributed formation method, and can also be formed in one step using a mask with multiple etching holes.

[0075] 5 and 6 , the slide body 10 in the biological slide 100 includes a substrate 1 and at least one adsorption structure layer 20 stacked on the surface of the substrate 1. The groove structure 4 is formed on the adsorption structure layer 20, wherein the first groove 4a and the second groove 4b extend into the adsorption structure layer 20 to different depths.

[0076] In some embodiments, the slide body 10 includes an adsorption structure layer 20. The adsorption structure layer 20 includes an adsorption layer 2 located on the surface of the substrate 1 and an isolation layer 3 located on the surface of the adsorption layer 2. The substrate 1 can be, for example, a silicon substrate (silicon material plate). The adsorption layer 2 is used to adsorb biological samples. For example, the adsorption layer 2 can be formed by an adsorption material that can adsorb biological samples. The groove structure 4 at least passes through the isolation layer 3 so that the adsorption layer 2 can be exposed by the groove structure 4, that is, the adsorption layer 2 constitutes the bottom of the groove structure 4, thereby achieving adsorption and fixation of the biological sample.

[0077] In some embodiments, when the thickness of different locations within the same isolation layer 3 is equal, i.e., the isolation layer 3 has a uniform thickness, in order to form groove structures 4 of varying depths, all or part of the groove structures 4 need to extend into the adsorption layer 2, but a portion of the adsorption layer 2 needs to be left to adsorb the biological sample. It is understood that the thickness of the isolation layer 3 can also be set to be non-uniform, so that the depth difference between the closest groove structures 4 can be designed by varying the thickness of the isolation layer 3.

[0078] In some embodiments, as shown in FIG5 , the first groove 4a may only penetrate the isolation layer 3, while the second groove 4b may penetrate the isolation layer 3 and then extend into the adsorption layer 2 of a certain thickness, thereby forming groove structures 4 of varying depths. It is understood that the portion of the groove structure 4 extending into the adsorption layer 2 can be extended to different thickness positions of the adsorption layer 2 as needed to form groove structures 4 of various depths. As shown in FIG6 , all groove structures 4 extend into the adsorption layer 2, for example, the first groove 4c and the second groove 4d both extend into the adsorption layer 2, and the first groove 4c and the second groove 4d extend to different depths of the adsorption layer 2 according to the depth difference, thereby forming groove structures 4 of varying depths.

[0079] In some embodiments, the adsorption material may include at least one of a metal nitride (e.g., titanium nitride), a metal oxide (e.g., titanium oxide, zirconium oxide, zinc oxide), and a metal (e.g., silver). For example, the adsorption material may be a metal nitride (e.g., titanium nitride) or a metal oxide (e.g., titanium oxide).

[0080] In some embodiments, the thickness of the adsorption layer 2 may be 20 nm to 500 nm, further 30 nm to 300 nm, further 50 nm to 100 nm. For example, the thickness of the adsorption layer 2 may be 20 nm, 30 nm, 40 nm, 50 nm, 60 nm, 70 nm, 80 nm, 90 nm, 100 nm, 120 nm, 150 nm, 180 nm, 200 nm, 250 nm, 300 nm, 350 nm, 400 nm, 450 nm, or 500 nm. When it is necessary to design groove structures 4 with various groove depths, the thickness of the adsorption layer 2 may be designed to be thicker.

[0081] In some embodiments, the isolation layer 3 is made of silicon oxide, silicon nitride, or other oxides or nitrides.

[0082] In some embodiments, the thickness of the isolation layer 3 may be 20 nm to 150 nm, and the thickness of the isolation layer 3 may be designed according to the actual depth of the groove structure 4 .

[0083] In some embodiments, a buffer layer 5 is provided between the substrate 1 and the adsorption layer 2. The buffer layer 5 may be made of silicon oxide, silicon nitride, or other oxides or nitrides. In some embodiments, a 20 nm to 1000 nm thick buffer layer 5, such as a SiO2 layer, may be deposited on the surface of the substrate 1. Adding the buffer layer 5 to the surface of the substrate 1 improves the adhesion of the adsorption layer 2 to the substrate 1, enhances the stability of the adsorption layer 2, and improves the quality of the biological slide 100.

[0084] The structure of the biological slide 100 provided in the embodiment of the present application has been comprehensively optimized. The first groove 4a and the second groove 4b, which are closest to each other, have different depths. Such groove structures 4 with different depths can greatly reduce the mutual interference between the optical signals of samples in adjacent reaction areas, increase the resolution of the fluorescence optical system, and improve the signal-to-noise ratio of the biological slide 100 during biochemical processing (such as sequencing). Moreover, by adjusting the depth and spacing of different groove structures 4, the density of the groove structures 4 can be increased, which is conducive to the preparation of a higher-throughput biological slide 100 and achieves the compatibility of higher signal quality and higher processing throughput. In addition, the biological slide 100 is reusable, effectively reducing the cost of use.

[0085] Referring to FIG. 7 , another embodiment of the present application provides another biological slide 200. The structure of the biological slide 200 is substantially the same as that of the biological slide 100 in the aforementioned embodiment, except that: the substrate 1 of the biological slide 200 is stacked with multiple adsorption structure layers 20 and 20 ′, and the first grooves 4 e and the second grooves 4 f extend into different layers of the adsorption structure layers 20 and 20 ′. Specifically, the first grooves 4 e extend into the adsorption structure layer 20 ′, and the second grooves 4 f extend into the adsorption structure layer 20, thereby forming first grooves 4 e and second grooves 4 f of different depths.

[0086] Specifically, the groove structures 4 on each adsorption structure layer 20 and 20' penetrate the outermost isolation layer 3', so that the adsorption layers 2 and 2' of different layers are exposed through the corresponding groove structures 4. In other words, the groove structures 4 corresponding to each adsorption structure layer 20 penetrate all other adsorption structure layers 20' located on the side of the adsorption structure layer 20 away from the substrate 1. As shown in Figure 7, the openings of the first groove 4e and the second groove 4f are both located on the surface of the outermost isolation layer 3' away from the substrate 1.

[0087] The biological slide 200 of this embodiment, by increasing the number of adsorption structure layers, facilitates the formation of groove structures 4 of varying depths, particularly groove structures 4 with significantly varying depths. This allows for a higher density of groove structures 4, further increasing biochemical processing throughput. Furthermore, the significant depth differences between groove structures 4 located on different adsorption structure layers further enhance the signal's ability to resist interference during detection, further improving signal quality.

[0088] Referring to FIG. 8 , the present embodiment further provides a method for preparing the aforementioned biological slide, which specifically includes the following steps:

[0089] Step S1 : forming a plurality of groove structures with openings located on the same side on a slide body of a biological slide, wherein the plurality of groove structures include a first groove and at least one second groove which is closest to the first groove and has a different depth from the first groove.

[0090] Step S2, forming a third groove adjacent to the first groove on the carrier body, the distance between the third groove and the first groove is greater than the distance between the second groove and the first groove, and the depth of the third groove can be the same as that of the first groove.

[0091] Wherein, in step S1, according to the synchronous forming and step-by-step forming of the first groove and the second groove, the following two specific implementation methods may be included.

[0092] The synchronous molding method specifically comprises forming the first groove and the second groove on the carrier body at the same time. It is understandable that when a third groove is required, the third groove can also be formed simultaneously with the first and second grooves. When more types of grooves need to be formed, synchronous molding can also be used.

[0093] Referring to FIG. 9 , the steps of simultaneously forming the first groove and the second groove on the wafer body specifically include the following steps:

[0094] In step S11 , a patterned film having a first etching hole and a second etching hole is formed on the surface of the wafer body.

[0095] Step S12, removing a portion of the wafer carrier body corresponding to the first etching hole to form a first groove on the wafer carrier body, and removing a portion of the wafer carrier body corresponding to the second etching hole to form a second groove on the wafer carrier body.

[0096] The step-by-step forming method specifically comprises the following steps: forming a first groove and a second groove on the carrier body respectively.

[0097] Referring to FIG. 10 , forming the first groove and the second groove on the wafer body specifically includes the following steps:

[0098] In step S11 ′, a patterned film having first etching holes is formed on the surface of the wafer body.

[0099] Step S12 ′: removing a portion of the wafer carrier body corresponding to the first etching hole to form a first groove on the wafer carrier body.

[0100] In step S13 ′, a patterned film having second etching holes is formed on the surface of the wafer body.

[0101] Step S14 ′: removing a portion of the wafer carrier body corresponding to the second etching hole to form a second groove on the wafer carrier body.

[0102] Referring to FIG. 11 , in some embodiments, the slide body of the biological slide includes a base layer and at least one adsorption structure layer located on the base layer. The method for preparing the biological slide specifically includes the following steps:

[0103] In step S101 , at least one adsorption structure layer is formed on the surface of a substrate.

[0104] Specifically, the adsorption structure layer includes an adsorption layer located on the surface of the substrate and an isolation layer located on the surface of the adsorption layer, and the adsorption layer is used to adsorb biological samples.

[0105] Step S102 : forming a plurality of groove structures on the adsorption structure layer, and making the first groove and the second groove of the plurality of groove structures extend into the adsorption structure layer at different depths.

[0106] Specifically, when the adsorption structure layer includes an adsorption layer and an isolation layer stacked together, the groove structure at least penetrates the isolation layer so that the adsorption layer is exposed from the groove structure. Specifically, the first groove only penetrates the isolation layer, and the second groove penetrates the isolation layer and then extends to a portion of the adsorption layer.

[0107] Referring to FIG. 12 , in another embodiment, when multiple adsorption structure layers are formed on a substrate, the method for preparing a biological slide includes the following steps:

[0108] Step S101 ′: forming a multi-layer adsorption structure layer on the surface of the substrate.

[0109] Step S102 ′: forming a groove structure on each adsorption structure layer, and each groove structure penetrates the isolation layer in the outermost adsorption structure layer, so that the adsorption layer in each adsorption structure layer is exposed through the corresponding groove structure.

[0110] That is, the plurality of groove structures penetrate the isolation layers in different layers, so that the adsorption layers in different layers are exposed through the corresponding groove structures.

[0111] The preparation method of the biological slide 100 (200) provided in the embodiment of the present application has been comprehensively optimized in terms of the preparation process. By depositing an adsorption layer 2 (2') such as titanium nitride and an isolation layer 3 (3') such as silicon oxide on a substrate 1, and then patterning the isolation layer 3 (3') through mask design and a stepwise exposure, development and etching method to form an array of groove structures 4 with different groove depths, two adjacent groove structures 4 have different groove depths, thereby improving the signal quality of the detection and being able to produce a biological slide with a higher groove density, thereby improving the processing throughput. The preparation method has a simple process, and the dimensional accuracy of the groove structures 4 with different groove depths is high, thereby improving the stability and quality of the biochemical processing process (such as sequencing process) of the biological slide 100 (200).

[0112] The following is a further description of the preparation method of the biological slide through specific examples.

[0113] Example 1

[0114] Please refer to FIG. 13A to FIG. 13F and FIG. 5 . When it is necessary to form an adsorption structure layer 20 , the method for preparing the biological slide 100 includes the following steps:

[0115] In step S11 a , as shown in FIG13A , an adsorption layer 2 is formed on the surface of the substrate 1 .

[0116] In this step, for example, an 8-inch or 12-inch silicon substrate (wafer), for example, a silicon substrate with a thickness of about 725 μm, can be provided. Of course, this embodiment does not specifically limit the substrate type and size, and can be selected and adjusted accordingly according to actual needs.

[0117] In this step, before forming the adsorption layer 2, the preparation method further includes:

[0118] A buffer layer 5 is formed on the surface of the substrate 1 , wherein the adsorption layer 2 is located on the surface of the buffer layer 5 .

[0119] In this embodiment, the buffer layer 5 is formed by depositing a buffer material on the surface of the substrate 1. The buffer material includes silicon oxide, silicon nitride, or other oxides or nitrides. In this step, as an optional embodiment, silicon oxide is deposited on the surface of the silicon substrate 1 by chemical vapor deposition to form a buffer layer 5 with a thickness ranging from 20nm to 1000nm. For example, a silicon dioxide layer with a thickness of about 35nm is deposited as the buffer layer 5. The selection and adjustment can be made accordingly according to actual needs. It is understandable that the buffer layer 5 can also be omitted from the substrate 1.

[0120] In this step, the method for forming the adsorption layer 2 includes:

[0121] An adsorbent material for adsorbing biological samples is deposited on the surface of the substrate 1 to form the adsorption layer 2. The adsorption material includes at least one of titanium nitride, titanium oxide, silver, zirconium oxide, zinc oxide, and the like, and may be, for example, a metal nitride (titanium nitride) or a metal oxide (titanium oxide). In this embodiment, the adsorption material may be TiN, and the adsorption layer is a TiN layer, but this is not limited to this and can be adjusted and selected based on actual needs or potential requirements.

[0122] In this step, an adsorption material for adsorbing biological samples is deposited on the surface of the buffer layer 5 to form an adsorption layer 2 for adsorbing biological samples with a thickness ranging from 20 nm to 500 nm. For example, a layer of titanium nitride adsorption material with a thickness of approximately 60 nm is deposited on the buffer layer 5 by physical vapor deposition.

[0123] In this embodiment, the adsorption layer can be embodied in the form of an adsorption film. If the thickness of the adsorption film is too thin, it will be difficult to implement its deposition process and there will be uniformity problems. If the thickness is too thick, it will affect the quality of the biochemical reaction (such as sequencing). Therefore, the thickness of the adsorption layer 2 can be controlled within the above-mentioned optional thickness range, thereby ensuring the process stability while ensuring the quality of the biochemical reaction.

[0124] In this embodiment, the biological sample may be, for example, DNA nanospheres, etc., but is not limited thereto, and may be selected and adjusted accordingly according to actual needs.

[0125] In step S12 a , as shown in FIG13A , an isolation layer 3 is formed on the surface of the adsorption layer 2 .

[0126] In this step, the isolation layer 3 is formed by depositing an isolation material on the surface of the adsorption layer 2 . The isolation material includes silicon oxide, silicon nitride, or other oxides or other nitrides.

[0127] In some embodiments, a silicon oxide material is deposited on the adsorption layer 2 for adsorbing biological samples to form an isolation layer 3 with a thickness ranging from 20 nm to 150 nm. For example, silicon dioxide with a thickness of about 45 nm is deposited as the isolation layer 3 by chemical vapor deposition. If the thickness of the isolation layer 3 is too thin, it will be easily corroded during the subsequent reaction process, which will affect the stability of the entire process and it will be difficult to prepare groove structures with different groove depths; if the thickness of the isolation layer 3 is too thick, it will affect the fluorescence brightness, resulting in a weak signal and will affect the replacement of reagents in the groove, affecting the quality of biochemical reactions (such as sequencing). Therefore, the thickness of the isolation layer 3 can be controlled within the above-mentioned optional thickness range, thereby ensuring process stability while ensuring the quality of the biochemical reaction.

[0128] In step S13 a , as shown in FIG. 13A and FIG. 13B , a first patterned film 60 having a first etching hole 61 is formed on the surface of the isolation layer 3 .

[0129] There are two patterning methods for the first patterned film 60, namely etching and imprinting. The so-called patterning refers to the arrangement of multiple grooves in a specific pattern. The patterned film is a film layer with a groove structure arranged in a specific pattern.

[0130] The method of forming the first patterned film 60 by etching specifically includes the following steps:

[0131] In the first step, as shown in FIG13A , an anti-reflection layer 40 is formed on the surface of the isolation layer 3 .

[0132] The anti-reflection layer 40 is formed by depositing an anti-reflection material on the surface of the isolation layer 3 . The anti-reflection material includes silicon oxynitride or a bottom anti-reflection coating (BARC).

[0133] In some embodiments, an anti-reflection material is deposited on the isolation layer 3 to form an anti-reflection layer 40 having a thickness ranging from 50 nm to 70 nm. The anti-reflection material may be SiON material, and the anti-reflection layer 40 may be a SiON layer. Of course, this embodiment does not specifically limit the type and thickness of the anti-reflection material layer, and both can be selected and adjusted accordingly according to actual needs.

[0134] In the second step, as shown in FIG. 13A , a photoresist layer 50 is formed on the surface of the anti-reflection layer 40 .

[0135] Before forming the photoresist layer 50 , an adhesion-promoting coating layer (not shown) needs to be formed on the surface of the anti-reflection layer 40 , and the photoresist layer 50 is located on the surface of the adhesion-promoting coating layer.

[0136] Specifically, a layer of HMDS is first spin-coated on the surface of the anti-reflection layer 40 as an adhesion-promoting coating, and then a photoresist is spin-coated on the surface of the adhesion-promoting coating to form the photoresist layer 50 .

[0137] In this step, as an optional implementation, the thickness of the photoresist layer 50 may be in the range of 250 nm to 350 nm.

[0138] In the third step, as shown in FIG. 13B , a first etching hole 61 is formed in the photoresist layer 50 to obtain a first patterned film 60 .

[0139] Specifically, a mask having corresponding first grooves (the structure of the first mask as shown in FIG3A ) is used, and a regularly arranged circular groove array pattern (i.e., the first etching holes 61 ) is formed on the photoresist layer 50 by performing exposure and development operations using a DUV photolithography machine.

[0140] The method for forming the first patterned film by using the imprint method specifically includes the following steps:

[0141] The first step is to form an embossed adhesive layer on the surface of the isolation layer.

[0142] In the second step, a first etching hole is formed in the embossed adhesive layer to obtain a first patterned film.

[0143] In step S14 a , as shown in FIG. 13C , at least the isolation layer 3 corresponding to the first etching hole 61 is removed to expose the adsorption layer 2 , thereby forming a first groove 4 a on the adsorption structure layer 20 .

[0144] In this embodiment, only the isolation layer 3 corresponding to the first etched hole 61 is removed, exposing the adsorption layer 2, thereby forming the first trench 4a. Since the first trench 4a is formed by removing only the isolation layer 3 in this embodiment, the thickness of the isolation layer 3 can be appropriately thickened according to actual needs to obtain a suitable trench depth.

[0145] It is understandable that when photoresist is used to form the first patterned film 60, the adhesion promoting coating and the anti-reflection layer 40 need to be removed when removing the isolation layer 3. In this embodiment, the adhesion promoting coating, the anti-reflection layer 40 and the isolation layer 3 are removed by dry etching.

[0146] In this step, as an optional embodiment, dry etching is performed on the anti-reflective layer 40 and the isolation layer 3 (primarily for forming the sidewalls of the groove structures 4) to form a plurality of groove structures 4 with a predetermined groove diameter on the anti-reflective layer 40 and the isolation layer 3. Specifically, the predetermined inner diameter of the groove structure 4 can range from 100 nm to 300 nm and can be adjusted and set accordingly according to actual needs.

[0147] In step S15 a , as shown in FIG. 13D and FIG. 13E , a second patterned film 30 having a second etching hole 31 is formed on the surface of the isolation layer 3 , and the second patterned film 30 is extended to the first groove 4 a .

[0148] During the second etching process, the second patterned film 30 needs to fill the first groove 4 a , so as to protect the first groove 4 a from being affected.

[0149] The method for forming the second patterned film 30 is the same as the method for forming the first patterned film 60, as described in step S13a above, and will not be described in detail here. The difference is that the grooves on the mask used to form the second patterned film 30 correspond to the second grooves, as shown in the structure of the second mask in FIG3B .

[0150] In step S16 a , as shown in FIG. 13F , the isolation layer 3 and a portion of the adsorption layer 2 corresponding to the second etching hole 31 are removed to expose the adsorption layer 2 , thereby forming a second groove 4 b on the adsorption structure layer 20 .

[0151] In step S17 a , referring to FIG. 5 , the second patterned film 30 is removed, wherein the first groove 4 a and the second groove 4 b have different groove depths, and the first groove 4 a and the second groove 4 b constitute the plurality of groove structures 4 .

[0152] Specifically, the second patterned film 30 and the first patterned film 60 are first removed by dry etching and wet etching, and then the anti-reflection layer 40 is etched away by dry etching. It is understood that the first patterned film 60 can also be removed before step S15. To simplify the process, this embodiment chooses to remove the two patterned films together at the end.

[0153] In this embodiment, the groove diameters of the first groove 4a and the second groove 4b are approximately the same, which can be about 200 nm. The distance between the first groove 4a and the second groove 4b is 715 nm. The depth of the first groove 4a can be 45 nm, and the depth of the second groove 4b can be 65 nm. The bottom of the groove is titanium nitride for adsorbing DNA nanoballs, and the groove spacing is silicon oxide for non-specific adsorption of DNA nanoballs.

[0154] The above steps form a groove structure 4 with two groove depths. It can be understood that when groove structures with more than two groove depths need to be formed, steps S16a to S18a can be repeated.

[0155] Example 2

[0156] The following describes in detail a method for preparing a biological slide using the imprinting method, which specifically includes the following steps:

[0157] Step S11b: providing a substrate.

[0158] Step S12b: depositing an adsorption material for adsorbing the biological sample on the surface of the substrate to form an adsorption layer.

[0159] In this embodiment, the adsorption layer can be directly deposited on the surface of the substrate without depositing a buffer layer. This step is substantially the same as the aforementioned step S11a. For details, please refer to the aforementioned step S11a and will not be described in detail here. It is understandable that this step can also form a buffer layer on the substrate. For details, please refer to the aforementioned step S11a and will not be described in detail here.

[0160] Step S13b: depositing silicon oxide material on the surface of the adsorption layer to form an isolation layer.

[0161] This step is basically the same as the aforementioned step S12a. Please refer to the aforementioned step S12a for details and will not be described in detail here.

[0162] Step S14b: spin-coating a layer of adhesion promoter on the surface of the isolation layer, and then spin-coating a layer of embossing adhesive on the surface.

[0163] In step S15b, a pre-prepared working mold is used to emboss the embossed adhesive layer, and after curing and demolding processes, the pattern of the working mold is transferred to the embossed adhesive layer, thereby forming a first patterned film having a first etched hole.

[0164] In step S16b, the residual adhesive and the isolation layer in the first etching hole are removed by dry etching to form a first groove, and the adsorption layer at the bottom of the first groove is exposed.

[0165] In step S17b, a second patterned film having a second etching hole is formed on the surface of the isolation layer, and the second patterned film is extended to the first groove.

[0166] The method for forming the second patterned film is the same as the method for forming the first patterned film, see steps S14b and S15b above, and will not be described in detail here. The difference is that the grooves on the mask used to form the second patterned film correspond to the second grooves.

[0167] Step S18b, removing the isolation layer and part of the adsorption layer corresponding to the second etching hole to expose the adsorption layer, thereby forming a second groove on the adsorption structure layer, and finally removing the first patterned film and the second patterned film. The groove depths of the first groove and the second groove obtained are different, and the first groove and the second groove constitute the multiple groove structures.

[0168] In this embodiment, the groove diameters of the first groove and the second groove are approximately the same, which can be about 200 nm. The distance between the first groove and the second groove is 715 nm. The depth of the first groove can be 45 nm, and the depth of the second groove can be 65 nm. The bottom of the groove is titanium nitride for adsorbing DNA nanoballs, and the groove spacing is silicon oxide for non-specific adsorption of DNA nanoballs.

[0169] In this embodiment, the structure and size of the biological slide can refer to the structure and size involved in the method for preparing the biological slide in the first embodiment above, and thus will not be described in detail.

[0170] Example 3

[0171] 14A to 14E , together with FIG. 6 , when an adsorption structure layer 20 needs to be formed and the first groove also extends into the adsorption layer 2 , the method for preparing the biological slide 100 includes the following steps:

[0172] Step S21, as shown in FIG14A, is the same as the manufacturing method of the aforementioned step S11a. Please refer to the aforementioned step S11a and will not be described in detail here.

[0173] Step S22, as shown in Figure 14A, is identical to the fabrication method of step S12a described above. (Please refer to step S12a above and will not be further elaborated here.) The difference is that, because the first groove will subsequently extend into a portion of the adsorption layer 2, the thickness of the isolation layer 3 does not need to be excessively thick in this step. In this embodiment, the thickness of the isolation layer 3 can be 25 nm.

[0174] Step S23, as shown in FIG. 14A and FIG. 14B, is the same as the manufacturing method of the aforementioned step S13a. Please refer to the aforementioned step S13a and will not be described in detail here.

[0175] Step S24 , as shown in FIG14C , differs from the aforementioned step S14a in that this step requires removing the isolation layer 3 and also requires removing a portion of the adsorption layer 2 so that the first groove 4 c extends to the adsorption layer 2 .

[0176] In this embodiment, after etching away the isolation layer 3 , the adsorption layer 2 also needs to be etched away by about 20 nm.

[0177] Step S25, as shown in FIG. 14D and FIG. 14E, is the same as the manufacturing method of the aforementioned step S15a. Please refer to the aforementioned step S15a and will not be described in detail here.

[0178] Step S26, as shown in FIG14F, is the same as the manufacturing method of the aforementioned step S16a. Please refer to the aforementioned step S16a and do not repeat it here. It can be understood that the second groove 4d formed in this step has a different groove depth than the first groove 4c. The first groove 4c and the second groove 4d constitute a plurality of groove structures 4.

[0179] In this embodiment, during the formation of the second groove 4 d , after the isolation layer 3 is etched away, at least 40 nm of the adsorption layer 2 needs to be etched away.

[0180] Step S27, please refer to FIG6 , this step is the same as the manufacturing method of the aforementioned step S17a, please refer to the aforementioned step S17a, and will not be described in detail here.

[0181] In this embodiment, the inner diameters of the first groove 4c and the second groove 4d are approximately the same, which can be about 200 nm. The distance between the first groove 4c and the second groove 4d is 500 nm. The depth of the first groove 4a can be 45 nm, and the depth of the second groove 4b can be 65 nm. The bottom of the groove is titanium nitride for adsorbing DNA nanoballs, and the groove spacing is silicon oxide for non-specific adsorption of DNA nanoballs.

[0182] Example 4

[0183] 15A to 15F , when a multi-layer (e.g., two-layer) adsorption structure layer 20 is required, the method for preparing the biological slide 200 includes the following steps:

[0184] In step S31 , as shown in FIG15A , an adsorption layer 2 is formed on the surface of the substrate 1 .

[0185] This step is the same as the production method of the aforementioned step S11a. Please refer to the aforementioned step S11a and will not be described in detail here.

[0186] In this embodiment, a 35 nm thick layer of silicon dioxide is deposited as the buffer layer 5 , and a 60 nm thick layer of titanium nitride is deposited on the buffer layer 5 as the adsorption layer 2 .

[0187] In step S32 , as shown in FIG. 15A , an isolation layer 3 is formed on the surface of the adsorption layer 2 . The adsorption layer 2 and the isolation layer 3 constitute a first adsorption structure layer 20 .

[0188] This step is the same as the production method of the aforementioned step S11a. Please refer to the aforementioned step S11a and will not be described in detail here.

[0189] In this embodiment, the isolation layer 3 may be a 25 nm thick silicon dioxide layer.

[0190] In step S33 , as shown in FIG. 15A , steps S31 and S32 are repeated to form another adsorption layer 2 ′ and another isolation layer 3 ′ on the surface of the isolation layer 3 . The adsorption layer 2 ′ and the isolation layer 3 ′ constitute another adsorption isolation layer 20 ′.

[0191] In this embodiment, the adsorption layer 2 ′ may be a 20 nm thick titanium nitride layer, and the isolation layer 3 ′ may be a 45 nm thick silicon dioxide layer.

[0192] In step S34 , as shown in FIG. 15A and FIG. 15B , a first patterned film 60 having a first etching hole 61 is formed on the surface of the isolation layer 3 ′.

[0193] This step is the same as the production method of the aforementioned step S13a. Please refer to the aforementioned step S13a and will not be described in detail here.

[0194] In step S35 , as shown in FIG. 15C , at least the isolation layer 3 ′ corresponding to the first etching hole 61 is removed to expose the adsorption layer 2 ′, thereby forming a first groove 4 e on the adsorption structure layer 20 ′.

[0195] This step is the same as the production method of the aforementioned step S14a. Please refer to the aforementioned step S14a and will not be described in detail here.

[0196] In step S36 , as shown in FIG. 15D and FIG. 15E , a second patterned film 30 having a second etching hole 31 is formed on the surface of the isolation layer 3 ′, and the second patterned film 30 is extended to the first groove 4 e .

[0197] This step is the same as the production method of the aforementioned step S15a. Please refer to the aforementioned step S15a and will not be described in detail here.

[0198] In step S37 , as shown in FIG. 15F , the isolation layer 3 corresponding to the second etching hole 31 is removed to expose the adsorption layer 2 , thereby forming a second groove 4 f on the adsorption structure layer 20 .

[0199] This step is similar to the aforementioned step S16a, except that: due to the addition of an adsorption structure layer 20', the two groove structures have more room to adjust the groove depth, so this step does not require the removal of part of the adsorption layer 2. For the specific production method, please refer to the aforementioned step S16a and will not be described in detail here.

[0200] It is understandable that in this step, part of the adsorption layer 2 can also be removed simultaneously to produce a deeper second groove 4f.

[0201] In step S38 , referring to FIG. 7 , the first patterned film 60 and the second patterned film 30 are removed, wherein the first groove 4 e and the second groove 4 f have different groove depths, and the first groove 4 e and the second groove 4 f constitute the plurality of groove structures 4 .

[0202] In this embodiment, the groove diameters of the first groove 4e and the second groove 4f are approximately the same, which can be about 200 nm. The distance between the first groove 4e and the second groove 4f is 715 nm. The depth of the first groove 4e can be 45 nm, and the depth of the second groove 4f can be 90 nm. The bottom of the groove is titanium nitride for adsorbing DNA nanoballs, and the groove spacing is silicon oxide for non-specific adsorption of DNA nanoballs.

[0203] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present application and are not intended to limit the present application. Although the present application has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present application may be modified or replaced by equivalents without departing from the spirit and scope of the technical solutions of the present application.

Claims

1. A biological slide, characterized in that: It includes a wafer carrier body, which has a plurality of groove structures with openings located on the same side, and the plurality of groove structures include a first groove and at least one second groove that is closest to the first groove and has a different depth.

2. The biological slide according to claim 1, wherein A depth difference between the first groove and the second groove closest to the first groove is greater than or equal to 20 nm.

3. The biological slide according to claim 1, wherein The plurality of groove structures further include a third groove adjacent to the first groove. A distance between the third groove and the first groove is greater than a distance between the second groove and the first groove. The third groove and the first groove may have the same depth.

4. The biological slide according to any one of claims 1 to 3, wherein The depth of the groove structure is 20 nm to 150 nm.

5. The biological slide according to any one of claims 1 to 3, wherein: The carrier body includes a substrate and at least one adsorption structure layer located on the substrate. The groove structure is formed on the adsorption structure layer. The first groove and the second groove extend into the adsorption structure layer at different depths.

6. The biological slide according to claim 5, wherein The adsorption structure layer includes: an adsorption layer located on the surface of the substrate and an isolation layer located on the surface of the adsorption layer, wherein the adsorption layer is used to adsorb biological samples, the groove structure at least penetrates the isolation layer, and the adsorption layer is exposed by the groove structure.

7. The biological slide according to claim 6, wherein All or part of the groove structure extends into the adsorption layer.

8. The biological slide according to claim 5, wherein The biological slide includes multiple layers of the adsorption structure layer stacked together, and the first groove and the second groove extend into different layers of the adsorption structure layer.

9. The biological slide according to claim 1, wherein The inner diameter of the groove structure is 100 nm to 300 nm; and / or, The distance between the central axes of two adjacent groove structures is 200nm-1000nm.

10. The biological slide according to claim 6, wherein The adsorption layer comprises an adsorption material for adsorbing biological samples, wherein the adsorption material comprises at least one of metal nitride, metal oxide and metal, and / or, The material of the isolation layer includes at least one of metal nitride and metal oxide.

11. The biological slide according to claim 5, wherein A buffer layer is further provided between the substrate and the adsorption structure layer, and the material of the buffer layer includes at least one of oxide and nitride.

12. A method for preparing a biological slide, characterized in that: include: A plurality of groove structures with openings located on the same side are formed on the slide body of the biological slide, wherein the plurality of grooves The structure includes a first groove and at least one second groove which is closest to the first groove and has a different depth.

13. The method for preparing a biological slide according to claim 12, wherein: The step of forming a plurality of groove structures with openings located on the same side on the slide body of the biological slide comprises: forming the first groove and the second groove on the slide body at the same time; or The first groove and the second groove are respectively formed on the carrier body.

14. The method for preparing a biological slide according to claim 13, wherein: The step of simultaneously forming the first groove and the second groove on the wafer carrier body comprises: forming a patterned film having a first etching hole and a second etching hole on the surface of the carrier body; and The portion of the carrier body corresponding to the first etching hole is removed to form the first groove on the carrier body, and the portion of the carrier body corresponding to the second etching hole is removed to form the second groove on the carrier body.

15. The method for preparing a biological slide according to claim 13, wherein: The steps of forming the first groove and the second groove on the wafer carrier body include: forming a patterned film having first etching holes on the surface of the carrier body; removing a portion of the wafer carrier body corresponding to the first etching hole to form the first groove on the wafer carrier body; forming a patterned film having second etching holes on the surface of the carrier body; and A portion of the wafer carrier body corresponding to the second etching hole is removed to form the second groove on the wafer carrier body.

16. The method for preparing a biological slide according to claim 12, wherein: The preparation method further comprises: A third groove adjacent to the first groove is formed on the carrier body, the distance between the third groove and the first groove is greater than the distance between the second groove and the first groove, and the depth of the third groove and the first groove can be the same.

17. The method for preparing a biological slide according to claim 12, wherein: The step of forming a plurality of groove structures with openings located on the same side on the carrier body includes: forming at least one adsorption structure layer on the surface of the substrate; and The groove structure is formed on the adsorption structure layer, and the first groove and the second groove extend into the adsorption structure layer to different depths.

18. The method for preparing a biological slide according to claim 17, wherein: The step of forming at least one adsorption structure layer on the surface of the substrate comprises: forming an adsorption layer on the surface of the substrate, wherein the adsorption layer is used to adsorb biological samples; and forming an isolation layer on the surface of the adsorption layer, In the step of forming the groove structure on the adsorption structure layer, the groove structure at least penetrates the isolation layer, so that the adsorption layer is exposed from the groove structure.

19. The method for preparing a biological slide according to claim 12, wherein: A depth difference between the first groove and the second groove closest to the first groove is greater than or equal to 20 nm.

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