Multicolor resonant cavity light emitting diode based bioprobes and methods of making the same

Abstract

The application discloses a preparation method of a biological probe based on a multi-color resonant cavity light emitting diode, and comprises the following steps: bonding a plurality of single-color resonant cavity light emitting diodes with different light emitting wavelengths on a substrate to form a biological probe precursor, wherein the bonding of each single-color resonant cavity light emitting diode comprises the following steps: forming a first bonding metal layer on the substrate; sequentially forming a first reflecting layer and a second bonding metal layer on a sapphire epitaxial wafer; inverting the sapphire epitaxial wafer and bonding the first bonding metal layer and the second bonding metal layer to form a third bonding metal layer; etching the inverted sapphire epitaxial wafer to obtain an epitaxial layer; forming an insulating layer on the epitaxial layer; forming an N electrode and a P electrode on the insulating layer, which are suitable for applying voltage to the epitaxial layer to generate laser; forming a second reflecting layer on the insulating layer, which, together with the first reflecting layer, forms a resonant cavity for controlling the laser light emitting angle; sharpening the tip of the biological probe precursor to obtain the biological probe; and forming a biological encapsulating layer on the surface of the biological probe.

Description

Technical Field

[0001] This invention relates to a biological probe, and more particularly to a multicolor resonant cavity light-emitting diode-based biological probe for neural modulation applied in the field of optical genetics, and its preparation method. Background Technology

[0002] The basic principle of optogenetics is to extract light-sensitive gene fragments from prokaryotes, algae, fungi, bacteria, or some eukaryotes and transfer them into target cells via methods such as viruses, transgenes, or microinjection. After gene expression, these fragments generate opsin, also known as light-sensitive protein, on the cell membrane. Opsin is a photosensitive ion channel or pump that can regulate the concentration of ions across the cell membrane under different wavelengths of light stimulation, thereby changing the cell membrane potential and achieving the effects of cell activation (depolarization) or inactivation (polarization). In other words, by externally transfecting light-sensitive proteins onto the neurons to be studied, neuronal excitation or inhibition can be achieved through light irradiation. Because optogenetics avoids the disadvantages of electrical stimulation (low spatiotemporal precision, poor specificity for stimulating nerve cells, and lack of inhibitory function) and drug stimulation (long onset time and significant side effects), it currently has significant advantages in treating brain diseases such as depression, epilepsy, Alzheimer's disease, and anxiety.

[0003] Currently, LEDs are commonly used as stimulation light sources in the field of optogenetics. Typically, LED devices have a large emission angle, following a Lambertian distribution. With the help of a substrate, the emission angle of a conventional LED can reach 180°. This large emission angle can easily affect neighboring neurons during neuronal stimulation, and when performing multi-point stimulation, it can easily lead to optical crosstalk, thus causing deviations in the response of photosensitive proteins and resulting in inaccurate experimental results. Summary of the Invention

[0004] In view of this, the present invention provides a biological probe based on a multicolor resonant cavity light-emitting diode to effectively control the light emission angle of the light-emitting point and avoid optical crosstalk problems, thereby achieving precise control of neurons.

[0005] This invention provides a method for preparing a biological probe based on a multicolor resonant cavity light-emitting diode, comprising:

[0006] Multiple monochromatic resonant cavity light-emitting diodes with different emission wavelengths are bonded on a substrate to form a biological probe precursor. Bonding each monochromatic resonant cavity light-emitting diode includes: forming a first bonding metal layer on the substrate; forming a first reflective layer on a sapphire epitaxial wafer, the sapphire epitaxial wafer comprising, from bottom to top, a sapphire substrate, a buffer layer, a u-GaN layer, an n-GaN layer, a multiple quantum well active layer, an electron blocking layer, and a p-GaN layer; forming a second bonding metal layer on the first reflective layer; and bonding the sapphire epitaxial wafer with the second bonding metal layer to the substrate. The sapphire epitaxial wafer is flipped, and the first and second bonding metal layers are bonded together to form a third bonding metal layer; the flipped sapphire epitaxial wafer is etched to obtain an epitaxial layer formed on the first reflective layer; an insulating layer is formed on the epitaxial layer; an electrode layer is formed on the insulating layer, the electrode layer including N-electrodes and P-electrodes, the N-electrodes and P-electrodes being suitable for applying voltage to the epitaxial layer to generate laser light; a second reflective layer is formed on the insulating layer; wherein, the first reflective layer and the second reflective layer constitute a Fabry-Perot resonator suitable for controlling the emission angle of the laser light;

[0007] The tip of the biological probe precursor was sharpened to obtain a biological probe based on a multicolor resonant cavity light-emitting diode; and

[0008] A bio-encapsulation layer is formed on the surface of a bioprobe based on a multicolor resonant cavity light-emitting diode.

[0009] The present invention also provides a biological probe obtained by the above preparation method, comprising:

[0010] Substrate;

[0011] Multiple monochromatic resonant cavity light-emitting diodes with different emission wavelengths are formed on a substrate. The monochromatic resonant cavity light-emitting diodes include: a third bonding metal layer formed on the substrate; a first reflective layer formed on the third bonding metal layer; an epitaxial layer comprising a p-GaN layer, an electron blocking layer, a multi-quantum well active layer, and an n-GaN layer sequentially superimposed on the first reflective layer, wherein the electron blocking layer, the multi-quantum well active layer, and the n-GaN layer form a boss structure on the p-GaN layer; an insulating layer formed on the epitaxial layer, with an N-electrode via on the insulating layer located on the n-GaN layer and a P-electrode via on the insulating layer located on the p-GaN layer; an electrode layer including an N-electrode and a P-electrode, the N-electrode being formed within the N-electrode via and the P-electrode being formed within the P-electrode via, the N-electrode and P-electrode being adapted to apply a voltage to the epitaxial layer to generate laser light; and a second reflective layer formed on the insulating layer, wherein the second reflective layer is adapted to form a Fabry-Perot resonant cavity with the first reflective layer; and...

[0012] A bio-encapsulation layer is applied to multiple monochromatic resonant cavity light-emitting diodes with different emission wavelengths.

[0013] The biological probe based on a multicolor resonant cavity light-emitting diode provided in the above embodiments of the present invention employs an epitaxial layer to form the light-emitting region, and a first reflective layer and a second reflective layer are formed around the light-emitting region to form an optical resonant cavity. By adjusting the structure and thickness of the first reflective layer and the second reflective layer, the structure of the optical resonant cavity is changed, thereby controlling the light emission angle of the light-emitting region. By effectively controlling the light emission angle of the light-emitting region, optical crosstalk problems can be avoided, thereby achieving precise control of neurons. Attached Figure Description

[0014] Figure 1 This is a flowchart illustrating a method for preparing a biological probe based on a multicolor resonant cavity light-emitting diode according to an embodiment of the present invention.

[0015] Figures 2(a) to 2(c) are schematic diagrams of the preparation process of the epitaxial layer according to an embodiment of the present invention;

[0016] Figure 3 This is a cross-sectional schematic diagram of a monochromatic resonant cavity light-emitting diode according to an embodiment of the present invention;

[0017] Figure 4 This is a cross-sectional schematic diagram of the epitaxial layer according to an embodiment of the present invention;

[0018] Figure 5 A three-dimensional schematic diagram of a biological probe based on a multicolor resonant cavity light-emitting diode according to an embodiment of the present invention; and

[0019] Figure 6 This is a schematic diagram of the light emission angle of a biological probe based on a multicolor resonant cavity light-emitting diode according to an embodiment of the present invention.

[0020] [Explanation of Labels in the Attached Image]

[0021] 1-Substrate;

[0022] 2-Third bonding metal layer;

[0023] 3-First reflective layer;

[0024] 4-Epipolar layer;

[0025] 5-Insulation layer;

[0026] 6-Electrode layer;

[0027] 61-N electrode;

[0028] 62-P electrode;

[0029] 7-Second reflective layer;

[0030] 8-Bio-encapsulation layer;

[0031] 91-Sapphire substrate;

[0032] 92-Buffer layer;

[0033] 93-u-GaN layer;

[0034] 94-n-GaN layer;

[0035] 95-Multiple quantum well active layer;

[0036] 96-Electron blocking layer;

[0037] 97-GaN layer;

[0038] 110 - First monochromatic resonant cavity light-emitting diode;

[0039] 120 - Second monochromatic resonant cavity light-emitting diode. Detailed Implementation

[0040] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to specific embodiments and the accompanying drawings. However, this invention can be implemented in different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided to make the invention thorough and complete, and to fully convey the scope of the invention to those skilled in the art. In the accompanying drawings, for clarity, the dimensions and relative dimensions of layers and regions may be exaggerated, and the same reference numerals denote the same elements throughout.

[0041] The terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the invention. The terms “comprising,” “including,” etc., as used herein indicate the presence of the stated features, steps, operations, and / or components, but do not exclude the presence or addition of one or more other features, steps, operations, or components.

[0042] In the field of optogenetics, LEDs offer many advantages, such as millisecond-level (ms) switching, long lifespan, and the ability to emit light across the entire visible spectrum. Therefore, they are frequently used as stimulation sources for neuronal modulation. However, currently used LED-integrated biological probes all exhibit a Lambertian light emission angle that follows a Lambertian distribution. For multi-point neuronal modulation, the array of light-emitting points easily leads to optical crosstalk. Furthermore, before light stimulation, the neurons under study must be transfected with proteins sensitive to specific wavelengths of light (photosensitive proteins). These photosensitive proteins typically have a broad response wavelength range and tend to attach to adjacent neurons during transfection. In such cases, the excessively large emission angle of traditional array-type biological probes can easily cause inaccurate experimental results.

[0043] In view of this, the present invention provides a biological probe based on a multicolor resonant cavity light-emitting diode to effectively control the light emission angle of the light-emitting point, thereby avoiding the optical crosstalk problem present in biological probes with array-type LED light-emitting points, and thus achieving precise control of neurons.

[0044] The structure of a resonant-cavity light emitting diode (RCLED) mainly includes a first reflective layer at the bottom and a second reflective layer at the top, formed by metal or a distributed Bragg reflector (DBR), as well as an active region located between the first and second reflective layers.

[0045] Figure 1 This is a flowchart illustrating a method for preparing a biological probe based on a multicolor resonant cavity light-emitting diode according to an embodiment of the present invention.

[0046] According to an exemplary embodiment of the present invention, the present invention provides a method for preparing a biological probe based on a multicolor resonant cavity light-emitting diode, referring to... Figure 1 As shown, it includes steps S01 to S10.

[0047] In step S01, a first bonding metal layer is formed on substrate 1.

[0048] According to an embodiment of the present invention, the material of the substrate 1 includes at least one of the following: silicon, SiC, diamond, polyethylene terephthalate, polyimide, parylene, polyurethane, and polydimethylsiloxane.

[0049] According to an embodiment of the present invention, a first bonding metal layer can be formed on a substrate 1 by means of electron beam evaporation, thermal evaporation or sputtering. The material of the first bonding metal layer includes at least one of the following: Au, Sn, In; for example, the first bonding metal layer can be formed by electron beam evaporation of Au / Sn.

[0050] In step S02, a first reflective layer 3 is formed on the sapphire epitaxial wafer.

[0051] According to an embodiment of the present invention, the sapphire epitaxial wafer includes a sapphire substrate 91, a buffer layer 92, a u-GaN layer 93, an n-GaN layer 94, a multi-quantum-well active layer 95, an electron blocking layer 96, and a p-GaN layer 97.

[0052] According to an embodiment of the present invention, the n-GaN layer 94 can be Si-doped AlGaN, wherein the Si doping concentration is approximately 2 × 10⁻⁶. 19 cm -3 The multi-quantum-well active layer is made of Al. x Ga1-x N / GaN or In y Ga 1-y N / GaN, 0 < x < 1, 0 < y < 1. For example, the multi - quantum well active layer 95 can be 9 pairs of alternately arranged In 0.17 Ga 0.83 N / GaN; the material of the electron blocking layer 96 can be p - Al 0.1 Ga 0.9 N. For example, the material of the electron blocking layer 96 can be Mg - doped AlGaN; the p - GaN layer 97 can be Mg - doped AlGaN or Mg - doped GaN, where the doping concentration of Mg is about 3.6×10 19 cm -3 .

[0053] According to an embodiment of the present invention, a first reflective layer 3 is formed on the p - GaN layer 97 of the sapphire epitaxial wafer; the first reflective layer 3 is a distributed Bragg reflector formed by periodically stacking high and low refractive index materials in an alternating form. For example, the first reflective layer 3 includes periodically stacked TiO2 and SiO2, and the number of stacking periods can be 15, but is not limited to 15; the number of stacking periods of the first reflective layer 3 can be adjusted according to actual needs.

[0054] In step S03, a second bonding metal layer is formed on the first reflective layer 3.

[0055] According to an embodiment of the present invention, the second bonding metal layer is formed of the same material as the first bonding metal layer.

[0056] In step S04, the sapphire epitaxial wafer formed with the second bonding metal layer is flipped, and the first bonding metal layer and the second bonding metal layer are bonded to form a third bonding metal layer 2.

[0057] According to an embodiment of the present invention, the first bonding metal layer formed on the substrate 1 and the second bonding metal layer formed on the first reflective layer 3 are bonded to form a third bonding metal layer 2, so that the first reflective layer 3 is transferred to the substrate 1. The methods for realizing the bonding of the first bonding metal layer and the second bonding metal layer include but are not limited to thermocompression bonding, diffusion bonding, and eutectic bonding.

[0058] In step S05, the flipped sapphire epitaxial wafer is etched to obtain the epitaxial layer 4 formed on the first reflective layer 3.

[0059] Figures 2(a) - 2(c) are schematic diagrams of the preparation process of the epitaxial layer according to an embodiment of the present invention.

[0060] According to an embodiment of the present invention, a laser is used to peel off the sapphire substrate 91 and buffer layer 92 of the sapphire epitaxial wafer. Referring to Figures 2(a) to 2(b), the sapphire substrate 91 is first peeled off using a laser, such as a 248nm KrF excimer laser or a 355nm third-harmonic Nd:YVO4 laser. It should be noted that during the laser peeling of the sapphire substrate 91, other layers besides the sapphire substrate 91 are sacrificed, which also leads to the peeling off of the buffer layer 92, exposing the u-GaN layer 93.

[0061] According to an embodiment of the present invention, the u-GaN layer 93 is treated with chemical mechanical polishing. Specifically, the u-GaN layer 93 is removed by chemical mechanical polishing to expose the n-GaN layer 94 and to make the surface of the n-GaN layer 94 smoother.

[0062] According to an embodiment of the present invention, referring to Figures 2(b) to 2(c), the n-GaN layer 94, the multi-quantum well active layer 95, the electron blocking layer 96, and the p-GaN layer 97 are etched by ICP (inductively coupled plasma etching) to expose the first reflective layer 3, and the electron blocking layer 96, the multi-quantum well active layer 95, and the n-GaN layer 94 form a boss structure on the p-GaN layer 97.

[0063] In step S06, an insulating layer 5 is formed on the epitaxial layer 4.

[0064] According to an embodiment of the present invention, SiO2 or Si3N4 is deposited on the epitaxial layer 4 as an insulating layer 5 using PECVD (plasma-enhanced chemical vapor deposition), and the insulating layer 5 can serve as a sidewall protective layer.

[0065] In step S07, an electrode layer 6 is formed on the insulating layer 5. The electrode layer 6 includes an N electrode 61 and a P electrode 62.

[0066] According to an embodiment of the present invention, firstly, an electrode pattern is defined on the insulating layer 5 using photolithography. Then, an N-electrode via is formed on the insulating layer 5 located on the n-GaN layer 94 using a BOE etching solution (a mixed solution of hydrofluoric acid and ammonium fluoride), and a P-electrode via is formed on the insulating layer 5 located on the p-GaN layer 97. Then, an N-electrode 61 is formed in the N-electrode via using electron beam evaporation, thermal evaporation, or sputtering, and a P-electrode 62 is formed in the P-electrode via. The material of the electrode layer 6 includes at least one of the following: Au, Ag, Cu, Pt, Cr, Ni, Al, Ti. For example, the electrode layer 6 can be a Cr / Al / Ti / Au alloy.

[0067] In step S08, a second reflective layer 7 is formed on the insulating layer 5 to obtain a monochromatic resonant cavity light-emitting diode.

[0068] According to an embodiment of the present invention, the second reflective layer 7 is a distributed Bragg reflector formed by periodically stacking high and low refractive index materials in an alternating manner. The materials of the first reflective layer 3 and the second reflective layer 7 may be the same or different, and the thicknesses of the first reflective layer 3 and the second reflective layer 7 may be the same or different. For example, the second reflective layer 7 includes periodically stacked TiO2 and SiO2, and the number of stacking periods may be 15, but is not limited to 15. The number of stacking periods of the second reflective layer 7 can be adjusted according to actual needs.

[0069] According to an embodiment of the present invention, the first reflective layer 3 and the second reflective layer 7 constitute a Fabry-Perot resonant cavity to regulate the light emission angle of the light emitted from the light-emitting region formed by the epitaxial layer 4. By effectively controlling the light emission angle of the light-emitting point, the light crosstalk problem present in the biological probe of the array LED light-emitting point can be avoided, thereby achieving precise regulation of neurons. In addition, the second reflective layer 7 can also serve as an insulating protective layer for the electrode layer 6.

[0070] In step S09, the tip of the biological probe precursor is sharpened to obtain a biological probe based on a multicolor resonant cavity light-emitting diode.

[0071] It should be noted that multiple monochromatic resonant cavity light-emitting diodes with different emission wavelengths bonded to substrate 1 constitute the biological probe precursor.

[0072] According to an embodiment of the present invention, a laser is used to define the shape of the biological probe precursor and to sharpen the tip of the biological probe precursor.

[0073] In step S10, a bio-encapsulation layer 8 is formed on the surface of the bio-probe based on a multicolor resonant cavity light-emitting diode.

[0074] According to an embodiment of the present invention, before forming the bio-encapsulation layer 8, the substrate is thinned by polishing, and then the bio-probe is bonded to the reserved metal pad by bonding technology. The bonding technology used includes, but is not limited to, solder paste, silver paste or In pillars.

[0075] According to embodiments of the present invention, the bio-encapsulation layer 8 is formed by spin coating or deposition. The material of the bio-encapsulation layer 8 includes at least one of the following: polydimethylsiloxane, polyethylene terephthalate, parylene, negative epoxy photoresist, epoxy resin, polyisobutylene, silicone, and hydrogel.

[0076] Figure 3 This is a cross-sectional schematic diagram of a monochromatic resonant cavity light-emitting diode according to an embodiment of the present invention. Figure 4 This is a cross-sectional schematic diagram of the epitaxial layer according to an embodiment of the present invention. Figure 5This is a three-dimensional schematic diagram of a biological probe based on a multicolor resonant cavity light-emitting diode according to an embodiment of the present invention.

[0077] According to an exemplary embodiment of the present invention, the present invention provides a biological probe based on a multicolor resonant cavity light-emitting diode, with reference to... Figures 3-5 As shown, it includes:

[0078] Substrate 1;

[0079] Multiple monochromatic resonant cavity light-emitting diodes with different emission wavelengths are formed on a substrate 1. Each monochromatic resonant cavity light-emitting diode includes: a third bonding metal layer 2 formed on the substrate 1; a first reflective layer 3 formed on the third bonding metal layer 2; and an epitaxial layer 4, which includes a p-GaN layer 97, an electron blocking layer 96, a multi-quantum well active layer 95, and an n-GaN layer 94 sequentially covering the first reflective layer 3. The electron blocking layer 96, the multi-quantum well active layer 95, and the n-GaN layer 94 are located on the p-GaN layer 97. A boss structure is formed on the first reflective layer 3; an insulating layer 5 is formed on the epitaxial layer 4, with an N-electrode via on the insulating layer 5 located on the n-GaN layer 94 and a P-electrode via on the insulating layer 5 located on the p-GaN layer 97; an electrode layer 6 includes an N-electrode 61 and a P-electrode 62, with the N-electrode 61 formed in the N-electrode via and the P-electrode 62 formed in the P-electrode via; a second reflective layer 7 is formed on the insulating layer 5, wherein the second reflective layer 7 is adapted to form a Fabry-Perot resonant cavity with the first reflective layer 3; and

[0080] The bio-encapsulation layer 8 covers multiple monochromatic resonant cavity light-emitting diodes (RCLEDs) on the bio-probe with different emission wavelengths.

[0081] According to an embodiment of the present invention, the emission wavelength of the monochromatic resonant cavity light-emitting diode is 350~650nm. For example, the emission wavelength can be one of the following: 450nm, 505nm, 580nm, 640nm.

[0082] Figure 6 This is a schematic diagram of the light emission angle of a biological probe based on a multicolor resonant cavity light-emitting diode according to an embodiment of the present invention.

[0083] According to an embodiment of the present invention, reference Figure 6 As shown, the biological probe based on multicolor resonant cavity light-emitting diodes includes multiple monochromatic resonant cavity light-emitting diodes with different emission wavelengths, such as a first monochromatic resonant cavity light-emitting diode 110 and a second monochromatic resonant cavity light-emitting diode 120.

[0084] The biological probe based on a multicolor resonant cavity light-emitting diode provided in the above embodiments of the present invention employs an epitaxial layer to form the light-emitting region, and a first reflective layer and a second reflective layer are formed around the light-emitting region to form an optical resonant cavity. By adjusting the structure and thickness of the first reflective layer and the second reflective layer, the structure of the optical resonant cavity is changed, thereby controlling the light emission angle of the light-emitting region. By effectively controlling the light emission angle of the light-emitting region, the optical crosstalk problem present in array-type light-emitting probes can be avoided, thereby achieving precise control of neurons.

[0085] The biological probe based on multicolor resonant cavity light-emitting diodes provided by the above embodiments of the present invention includes multiple monochromatic resonant cavity light-emitting diodes with different emission wavelengths. The emission wavelength and number of the stimulation light source can be selected according to actual needs, so as to study the effects of different wavelengths of light on animal neurons while precisely controlling neurons, thus providing a method for precise research on neural activity.

[0086] The use of ordinal numbers such as "first," "second," "third," etc., in the specification and claims to modify the corresponding elements does not imply that the element has any ordinal number, nor does it represent the order of one element with another element, or the order of manufacturing methods. The use of these ordinal numbers is only to enable a named element to be clearly distinguished from another element with the same name.

[0087] The specific embodiments described above further illustrate the purpose, technical solution, and beneficial effects of the present invention. It should be understood that the above descriptions are merely specific embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A method for preparing a biological probe based on a multicolor resonant cavity light-emitting diode, characterized in that, include: Multiple monochromatic resonant cavity light-emitting diodes with different emission wavelengths are bonded on the substrate (1) to form a biological probe precursor. The bonding of each monochromatic resonant cavity light-emitting diode includes: A first bonding metal layer is formed on the substrate (1); A first reflective layer (3) is formed on a sapphire epitaxial wafer, which, from bottom to top, includes a sapphire substrate (91), a buffer layer (92), a u-GaN layer (93), an n-GaN layer (94), a multi-quantum well active layer (95), an electron blocking layer (96), and a p-GaN layer (97). A second bonding metal layer is formed on the first reflective layer (3); The sapphire epitaxial wafer with the second bonding metal layer is flipped, and the first bonding metal layer and the second bonding metal layer are bonded to form a third bonding metal layer (2); The sapphire epitaxial wafer after being etched is used to obtain an epitaxial layer (4) formed on the first reflective layer (3); An insulating layer (5) is formed on the epitaxial layer (4); An electrode layer (6) is formed on the insulating layer (5), the electrode layer (6) including an N electrode (61) and a P electrode (62), the N electrode (61) and the P electrode (62) being adapted to apply a voltage to the epitaxial layer (4) to generate laser light; A second reflective layer (7) is formed on the insulating layer (5); wherein the first reflective layer (3) and the second reflective layer (7) constitute a Fabry-Perot resonant cavity, the Fabry-Perot resonant cavity being suitable for controlling the emission angle of the laser to avoid optical crosstalk between multiple monochromatic resonant cavity light-emitting diodes, wherein the first reflective layer (3) and the second reflective layer (7) are configured to be periodically stacked in an alternating manner of high and low refractive index materials; The tip of the biological probe precursor is sharpened to obtain a biological probe based on a multicolor resonant cavity light-emitting diode; and A bio-encapsulation layer (8) is formed on the surface of the bio-probe based on a multicolor resonant cavity light-emitting diode.

2. The preparation method according to claim 1, characterized in that, The sapphire epitaxial wafer after etching and flipping yields an epitaxial layer (4) formed on the first reflective layer (3), comprising: The sapphire substrate (91) and the buffer layer (92) of the sapphire epitaxial wafer are separated by a laser. The u-GaN layer was treated with chemical mechanical polishing (93); The n-GaN layer (94), the multi-quantum well active layer (95), the electron blocking layer (96), and the p-GaN layer (97) are etched to expose the first reflective layer (3), and the electron blocking layer (96), the multi-quantum well active layer (95), and the n-GaN layer (94) are formed into a boss structure on the p-GaN layer (97).

3. The preparation method according to claim 1, characterized in that, The bonding methods for the first bonding metal layer and the second bonding metal layer include one of hot-press bonding, diffusion bonding, or eutectic bonding.

4. A biological probe based on a multicolor resonant cavity light-emitting diode obtained by the preparation method according to any one of claims 1 to 3, characterized in that, include: Substrate (1); Multiple monochromatic resonant cavity light-emitting diodes with different emission wavelengths are formed on the substrate (1), the monochromatic resonant cavity light-emitting diodes comprising: A third bonding metal layer (2) is formed on the substrate (1); A first reflective layer (3) is formed on the third bonded metal layer (2); Epitaxial layer (4), the epitaxial layer (4) includes a p-GaN layer (97), an electron blocking layer (96), a multi-quantum well active layer (95) and an n-GaN layer (94) sequentially covering the first reflective layer (3), wherein the electron blocking layer (96), the multi-quantum well active layer (95) and the n-GaN layer (94) form a boss structure on the p-GaN layer (97); An insulating layer (5) is formed on the epitaxial layer (4). An N-electrode via is provided on the insulating layer (5) located on the n-GaN layer (94), and a P-electrode via is provided on the insulating layer (5) located on the p-GaN layer (97). The electrode layer (6) includes an N electrode (61) and a P electrode (62), wherein the N electrode (61) is formed in the N electrode via and the P electrode (62) is formed in the P electrode via, and the N electrode (61) and the P electrode (62) are adapted to apply a voltage to the epitaxial layer (4) to generate laser light; A second reflective layer (7) is formed on the insulating layer (5), wherein the second reflective layer (7) is adapted to form a Fabry-Perot resonant cavity with the first reflective layer (3); wherein the first reflective layer (3) and the second reflective layer (7) are configured to be periodically stacked from alternating high and low refractive index materials; and A bio-encapsulation layer (8) is applied over multiple monochromatic resonant cavity light-emitting diodes with different emission wavelengths.

5. The biological probe based on a multicolor resonant cavity light-emitting diode according to claim 4, characterized in that, The emission wavelength of the monochromatic resonant cavity light-emitting diode is 350~650nm.

6. The biological probe based on a multicolor resonant cavity light-emitting diode according to claim 4, characterized in that, The material of the substrate (1) includes at least one of the following: Silicon, SiC, diamond, polyethylene terephthalate, polyimide, poly(p-xylene), polyurethane, polydimethylsiloxane.

7. The biological probe based on a multicolor resonant cavity light-emitting diode according to claim 4, characterized in that, The materials of the first reflective layer (3) and the second reflective layer (7) may be the same or different; The thickness of the first reflective layer (3) and the second reflective layer (7) may be the same or different.

8. The biological probe based on a multicolor resonant cavity light-emitting diode according to claim 4, characterized in that, The material of the multi-quantum-well active layer (95) is Al. x Ga 1-x N / GaN or In y Ga 1-y N / GaN, 0 <x<1,0<y<1。 9. The biological probe based on a multicolor resonant cavity light-emitting diode according to claim 4, characterized in that, The materials of the N electrode (61) and the P electrode (62) include at least one of the following: Au, Ag, Cu, Pt, Cr, Ni, Al, Ti; The bio-encapsulation layer (8) is made of at least one of the following materials: polydimethylsiloxane, polyethylene terephthalate, poly(p-xylene), negative epoxy photoresist, epoxy resin, polyisobutylene, silicone, or hydrogel.

10. The biological probe based on a multicolor resonant cavity light-emitting diode according to claim 4, characterized in that, The bioprobe based on a multicolor resonant cavity light-emitting diode is suitable for detecting photosensitive proteins.

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