Excitation wavelength dependent protein crystal luminescent material as well as preparation method and application thereof

Abstract

The invention discloses an excitation wavelength-dependent protein crystal luminescent material as well as a preparation method and application thereof, and particularly relates to the field of luminescent materials. Comprising the following steps: mixing a protein solution with a protein crystallization precipitant, and performing crystal incubation to obtain a protein crystal; and adding a glutaraldehyde solution into the protein crystal for cross-linking to obtain the cross-linked protein crystal. The natural product protein is used as a raw material, is wide in source, low in cost, green and renewable, does not need organic synthesis, has excellent biocompatibility and biodegradability, and meets the green and environment-friendly requirements; different from a traditional luminescent material which needs to determine a luminescent group, the luminescent mechanism of the protein crystal luminescent material is that after protein monomers are assembled into a crystal, unsaturated bonds of protein molecules in the crystal form a luminescent center through spatial intermolecular interaction, TSIs with different intensities are formed, and the luminescent wavelengths are different.

Description

Technical Field

[0001] The present application relates to the field of luminescent materials, and in particular to an excitation wavelength-dependent protein crystal luminescent material and a preparation method and application thereof. Background Art

[0002] Multicolor luminescent materials with photoluminescent properties have broad application prospects in the fields of lighting, display, detection, anti-counterfeiting, information storage and bio-imaging, and have received widespread attention. Based on Kasha's rule, traditional fluorescent materials can only emit monochromatic fluorescence under excitation of a specific wavelength. In order to achieve multicolor luminescence, a common strategy is to mix luminescent materials with different luminescent colors. However, this strategy will also lead to problems such as uneven dispersion of fluorescent materials and reduced luminescence performance. Therefore, it is necessary to develop luminescent materials that can achieve multicolor luminescence characteristics in a single component.

[0003] Aggregate-induced emission (AIE) materials have the characteristics of excitation wavelength dependence. Their luminescent color can be tuned by changing the wavelength of the excitation light. They are simple, fast, sensitive, and non-invasive, and are ideal materials for achieving multicolor luminescence in a single component. However, most traditional AIE materials are synthesized through organic synthesis, and the synthesis process is relatively complicated and involves toxic chemical reagents. They are not environmentally friendly and difficult to degrade, which greatly limits their practical application value. Summary of the invention

[0004] The main purpose of this application is to provide an excitation wavelength-dependent protein crystal luminescent material and a preparation method and application, aiming to solve the problem that existing AIE materials are difficult to degrade.

[0005] To achieve the above-mentioned purpose, the present application provides a method for preparing an excitation wavelength-dependent protein crystal luminescent material, comprising: dissolving a salt or a polymer material in a buffer solution to obtain a protein crystal precipitant; mixing the protein solution with the protein crystal precipitant and incubating the crystals to obtain protein crystals; adding a glutaraldehyde solution to the protein crystals for cross-linking to obtain cross-linked protein crystals.

[0006] Optionally, the volume ratio of the protein solution to the protein crystallization precipitant is 1:1.

[0007] Optionally, the salt includes sodium chloride or lithium nitrate, the polymer material includes polyethylene glycol, and the buffer includes acetic acid-sodium acetate, PBS, tris or MES.

[0008] Alternatively, the protein comprises lysozyme, hemoglobin, catalase, chymotrypsin, concanavalin, pepsin or lactoglobulin.

[0009] Optionally, crystal cultivation is carried out in a crystallization incubator at an incubation temperature of 4°C-20°C.

[0010] Optionally, the glutaraldehyde solution includes a protein crystallization precipitant and glutaraldehyde; wherein the volume concentration of glutaraldehyde in the glutaraldehyde solution is 0.5%-2%, and the cross-linking time is 8h-20h.

[0011] Optionally, after crystal cultivation, the method further comprises: washing the cultivated product with a protein crystal precipitant.

[0012] Optionally, after adding glutaraldehyde solution to the protein crystals for cross-linking, the method further comprises: washing and drying the cross-linked product with triple distilled water.

[0013] To achieve the above objectives, the present application also provides an excitation wavelength-dependent protein crystal luminescent material, which is obtained by the above preparation method. The protein crystal luminescent material emits red light, green light or blue light under the excitation of excitation light with a wavelength of 280nm-650nm.

[0014] To achieve the above objectives, the present application also provides an application of the above excitation wavelength-dependent protein crystal luminescent material for cell multicolor imaging, cell multimodal imaging, LED luminescence or anti-counterfeiting detection.

[0015] Compared with the prior art, the present invention has the following beneficial effects: (1) The raw materials are green, renewable and degradable: natural product proteins are used as raw materials, which are widely available, low-cost, green and renewable, do not require organic synthesis, have excellent biocompatibility and biodegradability, and meet the needs of green environmental protection; (2) The preparation method is simple, reproducible, and easy to prepare on a large scale: the sample can be obtained by mixing the protein solution and the protein crystallization precipitant in an aqueous solution, centrifuging, washing, and drying. The overall preparation process is green and pollution-free, can be prepared on a large scale, and is conducive to subsequent industrialization; (3) The luminescence mechanism is clear and easy to control: Different from traditional luminescent materials that require the determination of luminescent groups, the luminescence mechanism of the protein crystal luminescent material of the present application is that after the protein monomers are assembled into crystals, the unsaturated bonds (C═O, C═C, C═N and C≡N) of the protein molecules in the crystals form luminescence centers through spatial intermolecular interactions (TSI). Different intensities of TSI have different luminescence wavelengths. Stronger TSI emits red-shifted light, while weaker TSI emits blue-shifted light. Therefore, the luminescence color can be regulated by adjusting the strength of TSI, which is conducive to the precise control of the luminescence color; (4) Wide range of applications: Protein crystals can adjust their luminescence color by excitation wavelength and can be used in multicolor cell imaging, multicolor LED, anti-counterfeiting and other fields. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 This is a multicolor luminescence image of the tetragonal lysozyme crystal obtained in Example 1 of the present application excited at different wavelengths under a confocal microscope; Figure 2 This is the emission spectrum of the tetragonal lysozyme crystals obtained in Example 1 of the present application; Figure 3 This is a multicolor luminescence image of the orthorhombic lysozyme crystal obtained in Example 2 of the present application excited at different wavelengths under a confocal microscope; Figure 4 This is the emission spectrum of the orthorhombic lysozyme crystals obtained in Example 2 of the present application; Figure 5 This is a multicolor luminescence image of the triclinic lysozyme crystal obtained in Example 3 of the present application under different wavelengths of excitation under a confocal microscope; Figure 6 This is the emission spectrum of the triclinic lysozyme crystals obtained in Example 3 of the present application; Figure 7 This is a multicolor luminescence image of the catalase crystal obtained in Example 4 of the present application under different wavelengths of excitation under a confocal microscope; Figure 8 This is a multicolor luminescence image of the concanavalin crystals obtained in Example 5 of the present application when excited at different wavelengths under a confocal microscope.

[0017] Fig. 9 This is the CIE colorimetric system of the tetragonal lysozyme crystals obtained in Example 1 of the present application.

[0018] The realization of the purpose, functional features and advantages of this application will be further explained in conjunction with embodiments and with reference to the accompanying drawings. DETAILED DESCRIPTION

[0019] In order to make the purpose, technical solutions and advantages of this application clearer, the technical solutions in this application will be clearly and completely described below in conjunction with the drawings in this application. Obviously, the described embodiments are part of the embodiments of this application, not all of them. Based on the embodiments in this application, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of this application.

[0020] The first embodiment of the present invention provides a method for preparing a protein crystal luminescent material that is dependent on excitation wavelength, such as Figure 1 As shown, the specific steps include: Step S1, mixing a protein solution with a protein crystal precipitant, and incubating the crystals to obtain protein crystals; The volume ratio of the protein solution to the protein crystallization precipitant is 1: 1. The protein includes lysozyme, hemoglobin, catalase, chymotrypsin, concanavalin, pepsin or lactoglobulin.

[0021] Specifically, in step S11, the protein powder is dissolved in a protein crystallization buffer to obtain a protein solution; wherein the concentration of the protein crystallization buffer is 0.1-0.5 M, the pH is 4-10, and it is far away from the isoelectric point of the protein; Step S12, using a protein crystallization kit to screen the crystallization conditions of the target protein, or using a protein crystallization database (PDB database: https: / / www.rcsb.org / ) to search for protein crystallization conditions, to obtain a crystallization precipitant for the target protein; specifically, the preparation method of the protein crystallization precipitant is: dissolving a salt or a polymer material in a buffer to obtain a protein crystallization precipitant; wherein the salt includes sodium chloride or lithium nitrate, and the polymer material includes polyethylene glycol; the buffer includes acetic acid-sodium acetate, PBS (phosphate buffered saline), tris (trishydroxymethylaminomethane) or MES (2-morpholinoethanesulfonic acid, 2-Morpholinoethanesulfonic Acid).

[0022] Among them, the preparation method of acetic acid-sodium acetate buffer solution (pH 4.5) is: take 18g of sodium acetate, add 9.8mL of glacial acetic acid, and then add water to dilute to 1000mL to obtain pH 4.5 acetic acid-sodium acetate buffer solution.

[0023] Step S13, the protein solution and the protein crystallization precipitant are mixed in equal volumes, and the mixture is centrifuged at 8000 rpm for 5 minutes (or filtered using a 0.22 μm filter membrane) to obtain a protein crystallization solution.

[0024] Step S14, placing the protein crystallization solution into a crystallization incubator and incubating it at 4°C-20°C to promote crystal growth; and washing the incubated product multiple times with a protein crystallization precipitant to remove uncrystallized protein monomers to obtain target protein crystals.

[0025] Step S2, adding glutaraldehyde solution to the protein crystals for cross-linking to obtain cross-linked protein crystals.

[0026] Specifically, glutaraldehyde solution is added to the protein crystals for cross-linking for 8h-20h; the cross-linked product is washed multiple times with triple distilled water to remove uncross-linked glutaraldehyde, and freeze-dried to obtain the final cross-linked protein crystals. The glutaraldehyde solution is obtained by diluting glutaraldehyde with a protein crystallization precipitant; wherein the volume concentration of glutaraldehyde in the glutaraldehyde solution is 0.5%-2%. The protein crystallization precipitant can promote protein crystallization to prepare protein crystals; glutaraldehyde can cross-link protein crystals to improve the stability of the crystals; the concentration of glutaraldehyde used for cross-linking is relatively low, 1%. If it is added directly, it will be locally cross-linked, so it is diluted to a low concentration by a protein crystallization precipitant.

[0027] A protein crystal luminescent material dependent on excitation wavelength is obtained by the above-mentioned preparation method. The protein crystal luminescent material emits red light, green light or blue light under the excitation of excitation light with a wavelength of 280nm-650nm.

[0028] An application of the above-mentioned excitation wavelength-dependent protein crystal luminescent material is used for cell multicolor imaging, cell multimodal imaging, LED luminescence or anti-counterfeiting detection.

[0029] Example 1 Preparation of tetragonal lysozyme (protein) crystal multicolor luminescent material Step S1, dissolving 40 mg of lysozyme powder in 1 mL, 0.1 M acetic acid-sodium acetate buffer (pH 4.6) to obtain a lysozyme solution; dissolving 80 mg of sodium chloride in 1 mL, 0.1 M acetic acid-sodium acetate buffer (pH 4.6) to obtain a lysozyme crystal precipitant; mixing the lysozyme solution and the lysozyme crystal precipitant in equal volumes, filtering through a 0.22 μm filter membrane to obtain a lysozyme crystal solution; placing the lysozyme crystal solution in a crystallization incubator, incubating at 20° C. to promote crystal growth for 48 hours, and obtaining tetragonal lysozyme crystals; collecting tetragonal lysozyme crystals by centrifugation (room temperature, 5000 rpm, 5 min), and washing the tetragonal lysozyme crystals three times with 0.1 M acetic acid-sodium acetate buffer (pH 4.6) containing 80 mg / mL sodium chloride to remove uncrystallized lysozyme monomers.

[0030] Step S2, adding 0.5% by volume of glutaraldehyde lysozyme crystal precipitant to the washed tetragonal lysozyme crystals for cross-linking for 8 hours to obtain cross-linked tetragonal lysozyme crystals; washing the cross-linked tetragonal lysozyme crystals three times with triple distilled water to remove uncross-linked glutaraldehyde, and freeze-drying to obtain the final tetragonal lysozyme crystals.

[0031] The multicolor luminescence properties of the tetragonal lysozyme crystals obtained in this embodiment were characterized using a confocal microscope and a fluorescence spectrometer. Figure 1 and Figure 2 As shown. Figure 1 It can be seen that the obtained tetragonal lysozyme crystals can be detected with bright blue, green and red fluorescence emissions in four fluorescence channels of the confocal microscope (λex=405nm, λem=425-475 nm; λex=488nm, λem=500-550nm; λex=561nm, λem=570-620 nm; λex=640 nm, λem=660-680nm), indicating that the synthesized lysozyme crystals have multicolor luminescence characteristics; Figure 2 Fluorescence spectra show that the prepared lysozyme crystals have excitation wavelength-dependent luminescence properties, and can emit red, green, and blue full-color fluorescence under the excitation of 280-550 nm excitation light. Therefore, excitation wavelength-dependent multicolor fluorescence emission is achieved in a single-component lysozyme crystal.

[0032] like Figure 3 As shown, the specific luminescent colors of tetragonal lysozyme crystals are: under the excitation of lasers with an excitation wavelength of 280nm-400nm, blue light can be emitted; under the excitation of lasers with an excitation wavelength of 410nm-500nm, green light can be emitted; and under the excitation of lasers with an excitation wavelength of 510nm-550nm, red light can be emitted. Example 2 Preparation of orthorhombic lysozyme crystal multicolor luminescent material Step S1, dissolving 20 mg of lysozyme powder in 1 mL, 0.1 M acetic acid-sodium acetate buffer (pH 4.6) to obtain a lysozyme solution; dissolving 80 mg of lithium nitrate in 1 mL, 0.1 M acetic acid-sodium acetate buffer (pH 4.6) to obtain a lysozyme crystal precipitant; mixing the lysozyme solution and the lysozyme crystal precipitant in equal volumes, filtering through a 0.22 μm filter membrane to obtain a lysozyme crystal solution; placing the lysozyme crystal solution in a crystallization incubator, incubating at 20° C. to promote crystal growth for 48 hours, and obtaining orthorhombic lysozyme crystals; collecting orthorhombic lysozyme crystals by centrifugation (room temperature, 5000 rpm, 5 min), and washing the orthorhombic lysozyme crystals three times with 0.1 M acetic acid-sodium acetate buffer (pH 4.6) containing 20 mg / mL lithium nitrate to remove uncrystallized lysozyme monomers.

[0033] Step S2, adding 0.5% by volume of glutaraldehyde lysozyme crystal precipitant to the washed orthorhombic lysozyme crystals for cross-linking for 8 hours to obtain cross-linked orthorhombic lysozyme crystals; using triple distilled water to wash the cross-linked orthorhombic lysozyme crystals three times, removing uncross-linked glutaraldehyde, and freeze-drying to obtain the final orthorhombic lysozyme crystals.

[0034] The multicolor luminescence properties of the orthorhombic lysozyme crystals obtained in this embodiment were characterized using a confocal microscope and a fluorescence spectrometer. Figure 4 and Figure 5 As shown. Figure 4 It can be seen that the obtained lysozyme crystals can be detected with bright blue, green and red fluorescence emissions in four fluorescence channels of the confocal microscope (λex=405nm, λem=425-475 nm; λex=488nm, λem=500-550nm; λex=561nm, λem=570-620 nm; λex=640 nm, λem=660-680nm), indicating that the synthesized lysozyme crystals have multicolor luminescence characteristics; Figure 5 Fluorescence spectra show that the prepared orthorhombic lysozyme crystals have luminescence characteristics that are dependent on the excitation wavelength, and can emit red, green, and blue full-color fluorescence under the excitation of 280-550 nm excitation light. Therefore, multicolor fluorescence emission dependent on the excitation wavelength is achieved in a single-component lysozyme crystal.

[0035] Example 3 Preparation of Triclinic Lysozyme Crystal Multicolor Luminescent Material Step S1, dissolving 40 mg of lysozyme powder in 1 mL, 0.1 M acetic acid-sodium acetate buffer (pH 4.6) to obtain a lysozyme solution; dissolving 80 mg of sodium chloride in 1 mL, 0.1 M acetic acid-sodium acetate buffer (pH 4.6) to obtain a lysozyme crystal precipitant; mixing the lysozyme solution and the lysozyme crystal precipitant in equal volumes, filtering through a 0.22 μm filter membrane to obtain a lysozyme crystal solution; placing the lysozyme crystal solution in a crystallization incubator, incubating at 40° C. to promote crystal growth for 48 hours, and obtaining triclinic lysozyme crystals; collecting triclinic lysozyme crystals by centrifugation (room temperature, 5000 rpm, 5 min), and washing the triclinic lysozyme crystals three times with 0.1 M acetic acid-sodium acetate buffer (pH 4.6) containing 80 mg / mL sodium chloride to remove uncrystallized lysozyme monomers.

[0036] Step S2, adding 0.5% by volume of glutaraldehyde lysozyme crystal precipitant to the cleaned triclinic lysozyme crystals for cross-linking for 8 hours to obtain cross-linked triclinic lysozyme crystals; using triple distilled water to wash the cross-linked triclinic lysozyme crystals three times, removing uncross-linked glutaraldehyde, and freeze-drying to obtain the final triclinic lysozyme crystals.

[0037] The multicolor luminescence properties of the obtained triclinic lysozyme crystals were characterized by confocal microscopy and fluorescence spectroscopy, such as Figure 6 and Figure 7 As shown. Figure 6It can be seen that the obtained lysozyme crystals can be detected with bright blue, green and red fluorescence emissions in four fluorescence channels of the confocal microscope (λex=405nm, λem=425-475 nm; λex=488nm, λem=500-550nm; λex=561nm, λem=570-620nm; λex=640 nm, λem=660-680nm), indicating that the synthesized lysozyme crystals have multicolor luminescence characteristics; Figure 7 Fluorescence spectra show that the prepared lysozyme crystals have excitation wavelength-dependent luminescence properties, and can emit red, green, and blue full-color fluorescence under the excitation of 280-550 nm excitation light. Therefore, excitation wavelength-dependent multicolor fluorescence emission is achieved in a single-component lysozyme crystal.

[0038] Example 4: Preparation of catalase crystal multicolor luminescent material Step S1, dissolving 8 mg / mL catalase powder in 1 mL, 0.1 M MES buffer (pH 6.3) to obtain a catalase solution; dissolving 20% ​​PEG 3350 in 1 mL, 0.1 M MES buffer (pH 6.3) to obtain a catalase crystallization precipitant; mixing the catalase solution and the catalase crystallization precipitant in equal volumes, filtering through a 0.22 μm filter membrane to obtain a catalase crystallization solution; placing the catalase crystallization solution in a crystallization incubator (20° C.) to promote crystal growth for 72 hours to obtain catalase crystals; collecting the catalase crystals by centrifugation (room temperature, 5000 rpm, 5 min), and washing the catalase crystals three times with 0.1 M MES buffer (pH 6.3) containing 20% ​​PEG 3350 to remove uncrystallized catalase monomers.

[0039] Step S2, adding a 0.5% by volume glutaraldehyde catalase crystal precipitant solution to the washed catalase crystals, cross-linking for 12 hours to obtain cross-linked catalase crystals; washing the cross-linked catalase crystals three times with triple distilled water to remove uncross-linked glutaraldehyde, and freeze-drying to obtain the final catalase crystals.

[0040] The multicolor luminescence properties of the catalase crystals obtained in this example were characterized using a confocal microscope. Figure 8 As shown. Figure 8It can be seen that the obtained catalase crystals can be detected in four fluorescence channels of the confocal microscope (λex=405nm, λem=425-475 nm; λex=488nm, λem=500-550nm; λex=561nm, λem=570-620 nm; λex=640 nm, λem=660-680nm) with bright blue, green, and red fluorescence emission, indicating that the synthesized catalase crystals have multicolor luminescence characteristics. Therefore, excitation wavelength-dependent multicolor fluorescence emission is achieved in a single-component catalase crystal.

[0041] Example 5 Preparation of Concanavalin Crystal Multicolor Luminescent Material Step S1, dissolving 20 mg / mL of concanavalin powder in 1 mL, 0.1 M acetic acid-sodium acetate buffer (pH 5.0) to obtain a concanavalin solution; dissolving 20% ​​PEG 3350 in 1 mL, 0.1 M acetic acid-sodium acetate buffer (pH 5.0) to obtain a concanavalin crystal precipitant; mixing the concanavalin solution and the concanavalin crystal precipitant in equal volumes, filtering through a 0.22 μm filter membrane to obtain a concanavalin crystal solution; placing the concanavalin crystal solution in a crystallization incubator (20°C) to promote crystal growth for 48 hours to obtain concanavalin crystals; collecting concanavalin crystals by centrifugation (room temperature, 5000 rpm, 5 min), and washing the concanavalin crystals three times with 0.1 M acetic acid-sodium acetate buffer (pH 4.6) containing 20% ​​PEG 3350 to remove uncrystallized concanavalin monomers.

[0042] Step S2, adding a 0.5% by volume glutaraldehyde concanavalin crystal precipitant solution to the washed concanavalin crystals, cross-linking for 12 hours to obtain cross-linked concanavalin crystals; washing the cross-linked concanavalin crystals three times with triple distilled water to remove uncross-linked glutaraldehyde, and freeze-drying to obtain the final concanavalin crystals.

[0043] The multicolor luminescence properties of the concanavalin crystals obtained in this example were characterized using a confocal microscope. Fig. 9 As shown. Fig. 9It can be seen that the obtained concanavalin crystals can be detected with bright blue, green, and red fluorescence emission in the four fluorescence channels of the confocal microscope ((λex=405nm, λem=425-475 nm; λex=488nm, λem=500-550nm; λex=561nm, λem=570-620 nm; λex=640 nm, λem=660-680nm)), indicating that the synthesized concanavalin crystals have multicolor luminescence characteristics. Therefore, excitation wavelength-dependent multicolor fluorescence emission is achieved in a single-component concanavalin crystal.

[0044] The above are only preferred embodiments of the present application, and are not intended to limit the patent scope of the present application. Any equivalent structure or equivalent process transformation made using the contents of the present application specification and drawings, or directly or indirectly applied in other related technical fields, are also included in the patent protection scope of the present application.

Claims

1. A method for preparing a protein crystal luminescent material that is dependent on excitation wavelength, characterized in that: include: Dissolving salt or polymer material in a buffer to obtain a protein crystal precipitant; mixing the protein solution with a protein crystallization precipitant and incubating the crystals to obtain protein crystals; A glutaraldehyde solution is added to the protein crystals for cross-linking to obtain cross-linked protein crystals.

2. The method for preparing the excitation wavelength-dependent protein crystal luminescent material according to claim 1, characterized in that: The volume ratio of the protein solution to the protein crystallization precipitant is 1:

1.

3. The method for preparing the excitation wavelength-dependent protein crystal luminescent material according to claim 1, characterized in that: The salt includes sodium chloride or lithium nitrate, and the polymer material includes polyethylene glycol; The buffer includes acetic acid-sodium acetate, PBS, tris or MES.

4. The method for preparing the excitation wavelength-dependent protein crystal luminescent material according to claim 1, characterized in that: The protein includes lysozyme, hemoglobin, catalase, chymotrypsin, concanavalin, pepsin or lactoglobulin.

5. The method for preparing the excitation wavelength-dependent protein crystal luminescent material according to claim 1, characterized in that: The crystal culture is carried out in a crystallization incubator at a temperature of 4°C-20°C.

6. The method for preparing the excitation wavelength-dependent protein crystal luminescent material according to claim 1, characterized in that: The glutaraldehyde solution comprises a protein crystallization precipitant and glutaraldehyde; Wherein, the volume concentration of glutaraldehyde in the glutaraldehyde solution is 0.5%-2%, and the cross-linking time is 8h-20h.

7. The method for preparing the excitation wavelength-dependent protein crystal luminescent material according to claim 1, characterized in that: After the crystal cultivation, the method further comprises: The cultured product is washed with a protein crystal precipitant.

8. The method for preparing the excitation wavelength-dependent protein crystal luminescent material according to claim 1, characterized in that: After adding glutaraldehyde solution to the protein crystals for cross-linking, the method further comprises: The cross-linked product was washed and dried with triple distilled water.

9. A protein crystal luminescent material that is dependent on excitation wavelength, characterized in that: The protein crystal luminescent material is obtained by the preparation method according to any one of claims 1 to 8, and emits red light, green light or blue light under the excitation of excitation light with a wavelength of 280nm-650nm.

10. A use of the excitation wavelength-dependent protein crystal luminescent material as claimed in claim 9, characterized in that: Used for multi-color cell imaging, multi-modal cell imaging, LED luminescence or anti-counterfeiting detection.

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