Method for manufacturing sporopollen phase identification slice under fluorescence
By mixing organic matter extract with glycerol during the preparation of pollen fossil thin sections and coating the outside with UV-curable adhesive to form a sealed space, the problem of fluorescence failure in traditional methods is solved, achieving efficient, economical and stable pollen phase analysis.
Patent Information
- Application Number
- CN202512039209.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-12-31
- Publication Date
- 2026-03-03
AI Technical Summary
Traditional thin-section preparation methods cannot simultaneously ensure the efficiency, safety, and economy of slide preparation while also taking into account the routine identification of pollen fossils and the analysis of pollen phases under fluorescent conditions. In particular, the organic components lose their fluorescent effect due to the absorption of ultraviolet light by the UV-cured adhesive.
An organic extract rich in fossil spores was mixed with glycerin and then coated with UV-curable adhesive along the outline of the coverslip to form a sealed space. The outer adhesive ring was cured by UV irradiation, avoiding direct contact between the UV-curable adhesive and the organic matter, thus ensuring that the fluorescence properties were not affected.
This method enables clear visualization of the morphology and structure of pollen fossils under a fluorescence microscope, meeting routine identification requirements. At the same time, it does not affect pollen phase analysis under a fluorescence microscope, exhibiting long-term stability and low cost, thus reducing material costs and operational risks.
Smart Images
Figure CN121595285A_ABST
Abstract
Description
Technical Field
[0001] This application relates to the field of microfossil analysis technology, and in particular to a method for preparing thin sections for identification under pollen fluorescence. Background Technology
[0002] In micropaleontological research, the identification of palynological fossils requires thin sections under an optical microscope. The preparation of palynological thin sections is fundamental to palynological phase (kerogen microstructure) analysis. This analysis uses transmitted light and fluorescence microscopy to observe the morphology, structure, and distribution characteristics of organic matter such as palynites to classify kerogen types. It is a key technology for evaluating the hydrocarbon generation potential and sedimentary environment of source rocks, with applications spanning multiple fields including oil and gas exploration, environmental science, and agricultural research.
[0003] However, traditional thin-section preparation methods have significant limitations: First, using glycerol gelatin, which requires heating to melt, as a medium results in a long preparation cycle, necessitates specific experimental facilities, incurs high material costs, and requires refrigeration. Furthermore, the prepared thin sections are prone to remelting and damage at higher ambient temperatures. Second, while using UV-curable adhesives directly mixed with organic matter extracts is efficient and low-cost, the UV-curable adhesive absorbs ultraviolet light, causing the organic components to lose their fluorescence, thus failing to meet the requirements for fluorescence analysis of the pollen phase (kerogen microstructure). Therefore, existing technologies cannot simultaneously ensure efficiency, safety, and economy in preparation while also accommodating routine identification of pollen fossils and precise analysis of the pollen phase (kerogen microstructure) under fluorescence conditions. A new, highly applicable, economical, and convenient process for preparing pollen fossil thin sections is urgently needed. Summary of the Invention
[0004] This application provides a method for preparing thin sections for fluorescent identification of pollen phases, which not only meets the requirements for the identification of pollen fossils, but also meets the requirements for the analysis of pollen phases (kerogen micro-components) under fluorescent conditions.
[0005] This application provides a method for preparing thin sections for pollen phase fluorescence identification, the method comprising: The organic matter extract rich in spore fossils was dropped onto a glass slide, and then glycerol was added to the organic matter extract on the glass slide and mixed to form a mixture. UV-curable adhesive is applied around the outline of the cover glass slide, and then the cover glass slide is placed over the predetermined area on the slide coated with the mixture, so that the mixture is confined within a sealed space formed by the slide, the cover glass slide and the surrounding UV-curable adhesive, to obtain a sheet to be cured. The sheet to be cured is subjected to ultraviolet irradiation to cure the ultraviolet curing adhesive, thereby obtaining the identification sheet.
[0006] Optionally, the amount of organic matter extract added is 1.0 μL to 2.0 μL.
[0007] Optionally, the amount of glycerol added is 0.03 mL to 0.07 mL.
[0008] Optionally, the amount of organic matter extract added is 1.5 μL, and the amount of glycerol added is 0.05 mL.
[0009] Optionally, the mixing is performed using a dissecting needle to ensure thorough blending of the glycerol with the organic extract and to remove air bubbles.
[0010] Optionally, the irradiation power of the ultraviolet light is 150W, and the wavelength range of the ultraviolet light is 365nm to 405nm.
[0011] Optionally, the ultraviolet irradiation time is 90s to 110s.
[0012] Optionally, the ultraviolet irradiation time is 99 seconds.
[0013] Optionally, the UV-curable adhesive comprises: isoborneol acrylate, hydroxypropyl methacrylate, acrylic acid, photoinitiator A, photoinitiator B, and silane.
[0014] Optionally, the ultraviolet-curable adhesive is UV adhesive K-306.
[0015] The technical solutions provided in this application have the following advantages compared with the prior art: This application provides a method for preparing a thin section for identifying pollen phases under fluorescence, achieving the dual requirements of pollen fossil identification and pollen phase analysis under fluorescence conditions through the following steps: First, an organic extract rich in pollen fossils is mixed with glycerol on a glass slide to form a mixture. Glycerol serves as a transparent medium, ensuring that the morphology and structure of the pollen fossils are clearly visible under an optical microscope, meeting the requirements for conventional identification. Second, an ultraviolet-curable adhesive is coated around the outline of a coverslip and then covered with the coverslip, confining the mixture within a sealed space formed by the glass slide, coverslip, and the surrounding ultraviolet-curable adhesive. This design strictly isolates the ultraviolet-curable adhesive from the outside of the mixture, preventing direct contact between it and the organic extract. This prevents the ultraviolet-curable adhesive from absorbing ultraviolet light and interfering with the fluorescence characteristics of the organic matter in the mixture during subsequent ultraviolet curing. Finally, the surrounding ultraviolet-curable adhesive is cured by ultraviolet irradiation, forming a firmly sealed identification thin section. This thin section ensures both the identification conditions of pollen fossils under transmitted light and the unaffected fluorescence display of the pollen phase (kerogen micro-components) under a fluorescence microscope, meeting the requirements for fluorescence analysis. Therefore, this process, through step design and material space separation, simultaneously meets the technical requirements for pollen fossil identification and pollen phase analysis under fluorescence conditions. Attached Figure Description
[0016] The accompanying drawings, which are incorporated in and form part of this specification, illustrate embodiments consistent with this application and, together with the description, serve to explain the principles of this application.
[0017] To more clearly illustrate the technical solutions in the embodiments of this application or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, for those skilled in the art, other drawings can be obtained based on these drawings without creative effort.
[0018] Figure 1 A schematic diagram of a thin section for a method of preparing a pollen phase fluorescence identification thin section provided in an embodiment of this application; Figure 2 Microscopic images (a) and (b) of the pollen fossils provided in Example 1 of this application, taken under transmitted light. Figure 3 Microscopic images of wood chips under transmitted light (a) and fluorescence (b) provided in Embodiment 2 of this application; Figure 4 Microscopic images (a) and (b) of organic matter fragments provided in Example 3 of this application under transmitted light; Figure 5 Microscopic images (a) and (b) of the pollen fossils provided in Comparative Example 1 of this application, taken under transmitted light. Detailed Implementation
[0019] To make the objectives, technical solutions, and advantages of the embodiments of this application clearer, the technical solutions of the embodiments of this application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this application, not all embodiments. Based on the embodiments of this application, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this application.
[0020] The range descriptions used herein, such as numerical ranges and proportional ranges, include all possible sub-ranges and single numerical values within that range. For example, the range descriptions of "1 to 6" or "1 to 6" cover all sub-ranges (such as 1 to 3, 2 to 5, etc.) and single numbers (such as 1, 2, 3, 4, 5, 6) between 1 and 6. Unless otherwise specified, the terms "including" and "contains" as used herein mean "including but not limited to"; relational terms such as "first" and "second" are used only to distinguish different entities or operations and do not imply an actual order or relationship; "and / or" indicates that multiple situations can exist individually or simultaneously; expressions such as "at least one," "multiple," and "at least one" refer to any combination of the corresponding objects, including combinations of single or multiple objects. The proportional relationships mentioned herein, such as mass ratios and molar ratios, should be understood as the correspondence between the first and second terms of a proportional formula, according to the order of description. The raw materials, reagents, instruments, and equipment used herein can all be obtained through commercial purchase or prepared using existing methods.
[0021] Figure 1 This is a schematic diagram of a thin section for a method of preparing a pollen-phase fluorescence identification thin section provided in an embodiment of this application.
[0022] like Figure 1 As shown in the embodiments of this application, a method for preparing thin sections for identification under pollen fluorescence is provided, the method comprising: S1. Add the organic matter extract rich in spore fossils to a glass slide, then add glycerol to the organic matter extract on the glass slide and mix to form a mixture; S2. Apply UV-curable adhesive around the outline of the coverslip, and then cover the predetermined area on the slide coated with the mixture with the coverslip, so that the mixture is confined within the sealed space formed by the slide, the coverslip and the surrounding UV-curable adhesive, to obtain the sheet to be cured. S3. Expose the film to be cured to ultraviolet light to cure the ultraviolet curing adhesive and obtain the test film.
[0023] The core of this application lies in providing a method for preparing pollen-phase fluorescence identification slides that integrates the advantages of sample fluorescence protection, rapid curing, and long-term stability. This method systematically solves the drawbacks of traditional processes through unique step design and parameter control.
[0024] Step S1: The preparation of the mixture aims to create a stable and fluorescence-compatible dispersion and observation medium for the pollen samples. Mixing the organic extract rich in pollen fossils with glycerol is crucial. Glycerol, a transparent liquid with moderate viscosity and suitable refractive index, serves several main functions: 1) As a dispersion medium, it ensures uniform suspension of pollen and other organic particles, preventing aggregation and sedimentation, facilitating microscopic observation of their morphology and distribution; 2) As a fluorescence observation medium, glycerol has extremely low fluorescence background and does not absorb excitation light, ensuring clear and interference-free visualization of the sample's fluorescence signal; 3) During mixing, stirring (e.g., using a dissecting needle) removes tiny air bubbles from the solution, preventing them from hindering microscopic observation and promoting thorough and uniform fusion of glycerol and the sample solution to form a stable mixture system.
[0025] Step S2: The core principle of constructing the curable sealing sheet is "physical isolation and peripheral sealing" to simultaneously achieve the dual goals of rapid curing and fluorescence protection. Specifically, UV-curable adhesive is first applied along the predetermined outline of the coverslip, and then the coverslip is placed over the area of the slide containing the mixture. This operation creates a sophisticated structure: the mixture is confined within a central sealed space formed by the slide, coverslip, and the uncured UV-curable adhesive on the periphery. Its core functions are: 1) Achieving fluorescence protection: By strictly confining the UV-curable adhesive to the periphery of the coverslip, it is completely physically isolated from the central glycerol-sample mixture. During subsequent curing, the UV light only acts on the peripheral colloid, avoiding direct contact between the curing adhesive components (especially photoinitiators) and the sample, thereby completely eliminating the fluorescence quenching effect caused by the mixing of colloid and sample in traditional methods, ensuring the clarity and accuracy of pollen fluorescence observation. 2) Constructing a sealed structure: This structure creates conditions for subsequent instantaneous curing and pre-forms a prototype of a sealed cavity that can isolate air and moisture for a long time, which is beneficial for the long-term preservation of the sample.
[0026] Step S3: The UV curing step is crucial for achieving high efficiency and stability in slide preparation. The prepared slide is then subjected to UV irradiation under specific conditions. The UV energy is absorbed by the outer UV-curing adhesive, triggering a rapid photopolymerization reaction within it, transforming it from a liquid to a solid state within tens of seconds. The purpose of this step is: 1) To achieve rapid and robust encapsulation: The UV curing process is rapid, significantly shortening the slide preparation time; the cured adhesive ring firmly bonds the coverslip to the slide and permanently seals the central mixture inside. 2) To impart environmental stability to the slide: The cured adhesive forms a hard sealing ring, preventing the prepared slide from being affected by changes in ambient temperature and humidity, avoiding the remelting or drying problems seen with glycerin gelatin slides, thus achieving long-term stable preservation of the slide.
[0027] In some embodiments, the amount of organic matter extract added is 1.0 μL to 2.0 μL.
[0028] In some embodiments, the amount of glycerol added is 0.03 mL to 0.07 mL.
[0029] The volume of organic matter extract added is limited to 1.0 μL to 2.0 μL, and the volume of glycerol added is limited to 0.03 mL to 0.07 mL. The volume of organic matter extract (e.g., 1.5 μL) ensures a sufficient and statistically significant number of pollen particles per unit area on the slide for identification and analysis. The volume of glycerol (e.g., 0.05 mL) needs to be sufficient to disperse the sample solution and form a liquid layer of suitable thickness to balance the requirements of microscope transmittance and three-dimensional observation of the sample. The set range represents an optimized balance between ensuring the quality of sample observation and avoiding excessive liquid leading to diffusion or excessive thickness affecting imaging. For example, the volume of organic matter extract added can be 1.0 μL, 1.25 μL, 1.5 μL, 1.75 μL, 2.0 μL, etc., and the volume of glycerol added can be 0.03 mL, 0.04 mL, 0.05 mL, 0.06 mL, 0.07 mL, etc.
[0030] In some embodiments, the amount of organic matter extract added is 1.5 μL, and the amount of glycerol added is 0.05 mL.
[0031] In some embodiments, a dissecting needle is used for stirring to ensure thorough mixing of the glycerol with the organic extract and to remove air bubbles.
[0032] In some implementations, the irradiation power of the ultraviolet light is 150W, and the wavelength range of the ultraviolet light is 365nm to 405nm.
[0033] In some implementations, the ultraviolet irradiation time is 90s to 110s.
[0034] In some implementations, the ultraviolet irradiation time is 99 seconds.
[0035] The UV irradiation power is limited to 150W, and the wavelength range is 365nm–405nm. These irradiation power (150W) and wavelength range (365–405nm) are efficient excitation conditions set for the photoinitiation characteristics of the selected UV-curable adhesive, providing sufficient and suitable light energy to initiate a rapid and complete polymerization reaction. The irradiation time (e.g., 99s) is a preferred value within the shortest effective time range (90–110s) required to ensure complete curing of the outer adhesive ring to achieve optimal adhesion and sealing strength, based on the above power and wavelength, balancing curing effect and sheet-making efficiency. For example, the UV irradiation time can be 90s, 95s, 100s, 105s, 110s, etc.
[0036] In some embodiments, the UV-curable adhesive comprises: isoborneol acrylate, hydroxypropyl methacrylate, acrylic acid, photoinitiator A, photoinitiator B, and silane.
[0037] In some implementations, the UV-curable adhesive is UV adhesive K-306.
[0038] This application uses acrylate resins such as isoborneol acrylate and hydroxypropyl methacrylate, combined with specific photoinitiators (A, B) and silanes to form a UV-curable adhesive (such as K-306) because such adhesives have the following characteristics: 1) strong adhesion to glass substrates (glass slides, coverslips); 2) fast curing speed and high hardness after curing under UV conditions; 3) their composition and curing characteristics are suitable for achieving peripheral coating and rapid photocuring processes, thus successfully constructing a thin sheet structure that is both firmly sealed and does not interfere with the fluorescence of the internal sample.
[0039] The method for preparing thin sections for pollen phase fluorescence identification proposed in this application exhibits a series of integrated and groundbreaking advantages compared to existing technologies. These advantages are rooted in its innovative core principle of "glycerol inner core - UV glue outer sealing" and are fully reflected in three aspects: technical performance, economic benefits, and operational practicality.
[0040] In terms of technical performance, this method achieves a balance between fluorescence fidelity, slide robustness, and long-term stability, overcoming the technical contradictions that traditional methods cannot achieve simultaneously. Traditional methods either use media that do not interfere with fluorescence (such as glycerol gelatin) but have slow curing and poor stability, or use fast-curing adhesives (such as UV-curing adhesives) but quench fluorescence due to direct contact. This application creatively achieves physical isolation through a step-by-step design that pre-mixes the sample with glycerol to form an observation core, and then applies UV-curing adhesive only to the periphery for sealing and curing. This core design completely avoids the influence of UV-curing adhesive and its curing process, thus perfectly preserving the inherent fluorescence characteristics of pollen and other organic matter. This allows for clear identification of the morphology, boundaries, and fluorescence reactions of microscopic components under a fluorescence microscope, laying a reliable foundation for accurate pollen phase analysis and kerogen type classification. At the same time, the outer rubber ring structure formed by rapid UV curing is robust and has excellent sealing properties, ensuring that the final sheet is completely resistant to changes in environmental temperature and humidity, never melting or drying out, and possesses the potential for permanent preservation. Its stability is far superior to traditional glycerol gelatin sheets that are easily melted by heat.
[0041] From an economic and operational perspective, this method combines low cost, high efficiency, high safety, and convenience, significantly lowering the barrier to professional analysis. The core materials required by the method (ordinary glycerin and conventional UV-curing adhesive) are inexpensive, readily available on the market, and require no special refrigeration. Compared to specialized glycerin gelatin, which requires refrigeration and is more expensive, this method significantly saves on material and warehousing costs. The slide preparation process eliminates the open flame heating step required by traditional methods, completely eliminating the corresponding safety risks and freeing slide preparation activities from being limited to specialized laboratories equipped with specific heating equipment, thus expanding the flexibility of application sites. The critical curing step requires only approximately 99 seconds of UV irradiation and can be batch-processed. Compared to traditional methods that require several hours of natural air drying, slide preparation efficiency is increased by orders of magnitude, enabling rapid fulfillment of the analytical needs for large batches of samples.
[0042] In summary, the advantage of this application lies in its ingenious and concise process design, which systematically integrates and surpasses the advantages of two traditional technical routes while avoiding their inherent defects. It successfully combines multiple ideal characteristics such as "non-destructive fluorescence observation," "instant curing and encapsulation," "strong environmental stability," "low economic cost," "high operational safety," and "excellent process efficiency." This not only provides a superior, reliable, and universally applicable standardized slide preparation tool for geoscience research and oil and gas exploration fields such as palynological analysis and source rock evaluation, but also, due to its convenience and economy, is expected to promote the widespread application of related microscopic analysis techniques in broader application scenarios such as environmental monitoring and agricultural research. Therefore, this method represents an innovative solution with significant advancements in both technical indicators and practical value.
[0043] The present application is further illustrated below with reference to specific embodiments. Experimental methods in the following embodiments that do not specify specific conditions are generally determined according to national standards / industry standards / the disclosure herein; if there are no corresponding national standards / industry standards / the disclosure herein, they are performed according to generally accepted international standards, conventional conditions, or conditions recommended by the manufacturer.
[0044] Example 1 This embodiment provides a method for preparing thin sections for identification under pollen fluorescence, the specific steps of which are as follows: S1. Preparation of the mixture: Place a glass slide on the table, and use a pipette to add 1.5 μL of organic matter extract rich in pollen fossils onto the slide. Stir and spread the extract with a dissecting needle to facilitate water evaporation. Then, add 0.05 mL of glycerol to the organic matter extract area on the slide, and slowly stir with a dissecting needle to ensure thorough mixing of the glycerol and organic matter extract, removing any air bubbles generated during the mixing process to form the mixture.
[0045] S2. Constructing the sheet to be cured: Apply a small amount of UV-curable adhesive K-306 around the outline of the coverslip, and then cover the area of the slide coated with the mixture with the coverslip. This operation confines the mixture within a sealed space formed by the slide, coverslip, and surrounding UV-curable adhesive, resulting in the sheet to be cured.
[0046] S3. UV Curing: Place the sheet to be cured horizontally under a UV lamp for irradiation. The UV lamp has an irradiation power of 150W, a wavelength range of 365nm to 405nm, and an irradiation time of 99s. After irradiation, the outer UV-curing adhesive is completely cured, resulting in a well-cured identification sheet.
[0047] Figure 2 Microscopic images (a) and (b) of the palynological fossils provided in Example 1 of this application, taken under transmitted light and fluorescence (scale bar 20 μm). Figure 2 As can be seen, the identification slides prepared according to the method of this embodiment, as verified by microscopic observation, have good light transmittance, are free of air bubbles, and have complete sealing. Under transmitted light, the morphology and structure of the pollen fossils are clearly visible; under fluorescence, the fluorescence reaction of the pollen fossils is obvious, and the boundaries are clear, fully meeting the technical requirements for the identification of pollen fossils and the fluorescence analysis of pollen phase (kerogen micro-components).
[0048] Example 2 This embodiment provides a method for preparing thin sections for pollen phase fluorescence identification, as detailed below: S1. Preparation of the mixture: Place a glass slide on the table, and use a pipette to add 1.0 μL of organic extract rich in sawdust onto the slide. Stir and spread the extract with a dissecting needle to facilitate water evaporation. Then, add 0.03 mL of glycerol to the organic extract area on the slide, and slowly stir with a dissecting needle to ensure thorough mixing of the glycerol and organic extract, removing any air bubbles generated during the mixing process to form the mixture.
[0049] S2. Constructing the sheet to be cured: Apply a small amount of UV-curable adhesive K-306 around the outline of the coverslip, and then cover the area of the slide coated with the mixture with the coverslip. This operation confines the mixture within a sealed space formed by the slide, coverslip, and surrounding UV-curable adhesive, resulting in the sheet to be cured.
[0050] S3. UV Curing: Place the sheet to be cured horizontally under a UV lamp for irradiation. The UV lamp has a power of 150W, a wavelength range of 365nm to 405nm, and an irradiation time of 90s. After irradiation, the outer UV-curing adhesive is completely cured, resulting in a well-cured identification sheet.
[0051] Figure 3Microscopic images (a) and (b) of wood chips provided in Example 2 of this application, taken under transmitted light and fluorescence (scale bar: 20 μm). Figure 3 As shown, the identification slide prepared according to the method of this embodiment, as verified by microscopic observation, has good light transmittance, is free of air bubbles, and has complete sealing. Under transmitted light, the morphology and structure of the sawdust are clearly visible; under fluorescence, the fluorescence reaction of the sawdust is obvious, with clear boundaries, fully meeting the technical requirements for fluorescence analysis of pollen phase (kerogen micro-components).
[0052] Example 3 This embodiment provides a method for preparing thin sections for pollen phase fluorescence identification, as detailed below: S1. Preparation of the mixture: Place a glass slide on the table, and use a pipette to add 2.0 μL of organic matter extract rich in organic matter fragments onto the slide. Stir and spread the extract with a dissecting needle to facilitate water evaporation. Then, add 0.07 mL of glycerol to the organic matter extract area on the slide, and slowly stir with a dissecting needle to ensure thorough mixing of the glycerol and organic matter extract, removing any air bubbles generated during the mixing process to form the mixture.
[0053] S2. Constructing the sheet to be cured: Apply a small amount of UV-curable adhesive K-306 around the outline of the coverslip, and then cover the area of the slide coated with the mixture with the coverslip. This operation confines the mixture within a sealed space formed by the slide, coverslip, and surrounding UV-curable adhesive, resulting in the sheet to be cured.
[0054] S3. UV Curing: Place the sheet to be cured horizontally under a UV lamp for irradiation. The UV lamp has an irradiation power of 150W, a wavelength range of 365nm to 405nm, and an irradiation time of 110s. After irradiation, the outer UV-curing adhesive is completely cured, resulting in a well-cured identification sheet.
[0055] Figure 4 Microscopic images (a) and (b) of the organic matter fragments provided in Example 3 of this application, taken under transmitted light and fluorescence (scale bar 20 μm). Figure 4 As shown, the identification slide prepared according to the method of this embodiment, as verified by microscopic observation, has good light transmittance, is free of bubbles, and has complete sealing. Under transmitted light, the morphology and structure of the organic matter fragments are clearly visible; under fluorescence, the fluorescence reaction of the organic matter fragments is obvious, with clear boundaries, fully meeting the technical requirements for fluorescence analysis of pollen phase (kerogen micro-components).
[0056] Comparative Example 1 This comparative example uses a traditional method to prepare pollen phase identification thin sections. The specific steps are as follows: S1. Use a pipette to take an appropriate amount of the organic extract rich in fossil spores and add it to a glass slide.
[0057] S2. Directly drop UV-curable adhesive K-306 into the organic extract on the glass slide, and then cover it with a coverslip.
[0058] S3. Place the glass slide covered with the coverslip under ultraviolet light for 2 minutes to cure the ultraviolet curing adhesive and obtain the identification slide.
[0059] Although this method has the advantages of convenient operation, fast curing and low cost, and the thin film produced is not sensitive to changes in environmental temperature and humidity, the UV curing adhesive is directly mixed with the organic matter extract rich in spores and pollen fossils and then cured by UV irradiation. During the curing process, the UV curing adhesive absorbs ultraviolet light, which causes the organic matter components to be disturbed and lose their original fluorescent properties.
[0060] Figure 5 Microscopic images (a) and (b) of the palynological fossils provided in Comparative Example 1 of this application, taken under transmitted light (scale bar: 20 μm). Figure 5 As shown, when the prepared thin sections were observed under a fluorescence microscope, the organic matter boundaries were blurred, and the fluorescence effect was weak or disappeared, which could not meet the requirements for fluorescence analysis of pollen phase (kerogen micro-components).
[0061] Comparative Example 2 This comparative example uses a traditional method to prepare pollen phase identification thin sections. The specific steps are as follows: S1. Place a glass slide on the table, and use a pipette to draw 1.5 μL of organic matter extract rich in pollen fossils and add it to the glass slide.
[0062] S2. Use a dissecting needle to take about 0.1 mL of solid glycerin gelatin, place it on a glass slide and make full contact with the organic matter extract.
[0063] S3. Heat the glass slide under an alcohol lamp to melt the glycerin gelatin, and stir it with a dissecting needle to fully mix it with the organic extract.
[0064] S4. Cover with a coverslip and place the sheet in a ventilated area for 2-3 hours to allow it to dry and harden completely.
[0065] The main advantage of this method is that glycerol gelatin does not absorb ultraviolet light, and the prepared thin films can be used for fluorescence analysis of pollen phases (kerogen microstructures).
[0066] However, this method has the following drawbacks: 1) High material cost, with glycerol gelatin being relatively expensive; 2) Stringent material storage conditions, unused glycerol gelatin needs to be refrigerated and stored, as it is prone to deterioration at room temperature; 3) Low sheet production efficiency, with the drying and curing time for the sheets being as long as 2-3 hours; 4) Safety risks and limited space during the sheet production process, requiring the use of open flames or similar heating equipment and operation in a specific laboratory environment; 5) Poor stability of the sheet product, as the finished product is prone to remelting at high ambient temperatures, leading to damage.
[0067] Furthermore, one or more technical solutions in the embodiments of this application have at least the following technical effects or advantages: In this embodiment, improvements are made to the preparation process and adhesive selection of pollen slices to differentiate them from traditional methods. This not only satisfies the identification of pollen fossils but also the analysis of pollen phases (kerogen microstructures) under fluorescent conditions.
[0068] In this embodiment, the pre-mixing of glycerol with the sample and the outer isolation encapsulation with UV-curable adhesive fundamentally avoid the quenching of the organic fluorescence signal by the curing adhesive components. Under a fluorescence microscope, the prepared thin sections show clear boundaries of pollen, sawdust, and other organic particles, with vivid fluorescence reactions, fully meeting the requirements for high-precision pollen phase analysis and kerogen microscopic component identification.
[0069] In this embodiment, a rigid sealing ring rapidly formed using UV-curable adhesive imparts extremely strong environmental robustness to the sheet. The resulting sheet exhibits excellent light transmittance, is bubble-free, and has a complete seal, unaffected by changes in ambient temperature and humidity. This overcomes the shortcomings of traditional glycerol gelatin sheets, which are prone to melting and drying, and enables long-term stable preservation of analytical samples.
[0070] In this embodiment, inexpensive and readily available common glycerin and general-purpose UV-curable adhesive (such as K-306) are used as the main materials, eliminating the need for expensive, cold-chain-required specialized glycerin gelatin. This not only significantly reduces the cost per slide preparation but also simplifies the storage and management requirements of experimental materials.
[0071] In this embodiment, the critical outer adhesive layer requires only about 99 seconds to cure, and it supports batch processing of thin films, greatly improving the overall efficiency of film production. This method is simple, requires no heating equipment, reduces operational complexity and safety risks, and allows for convenient high-quality film production in conventional laboratories and even field environments.
[0072] In the embodiments of this application, high-quality thin sections can be successfully produced for different types of organic samples (pollen fossils, wood chips, organic fragments), indicating that the method has wide applicability and good process repeatability, and provides a stable and reliable slide preparation solution for different research purposes.
[0073] The above description is merely a specific embodiment of this application, enabling those skilled in the art to understand or implement this application. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of this application. Therefore, this application is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features claimed herein.
Claims
1. A method for preparing thin sections for pollen phase fluorescence identification, characterized in that, The method includes: The organic matter extract rich in spore fossils was dropped onto a glass slide, and then glycerol was added to the organic matter extract on the glass slide and mixed to form a mixture. UV-curable adhesive is applied around the outline of the cover glass slide, and then the cover glass slide is placed over the predetermined area on the slide coated with the mixture, so that the mixture is confined within a sealed space formed by the slide, the cover glass slide and the surrounding UV-curable adhesive, to obtain a sheet to be cured. The sheet to be cured is subjected to ultraviolet irradiation to cure the ultraviolet curing adhesive, thereby obtaining the identification sheet.
2. The method according to claim 1, characterized in that, The amount of organic matter extract added is 1.0 μL to 2.0 μL.
3. The method according to claim 1, characterized in that, The amount of glycerol added is 0.03 mL to 0.07 mL.
4. The method according to claim 2 or 3, characterized in that, The amount of organic matter extract added is 1.5 μL, and the amount of glycerol added is 0.05 mL.
5. The method according to claim 1, characterized in that, The mixing process involves stirring with a dissecting needle to ensure thorough blending of the glycerol with the organic extract and to remove air bubbles.
6. The method according to claim 1, characterized in that, The irradiation power of the ultraviolet light is 150W, and the wavelength range of the ultraviolet light is 365nm to 405nm.
7. The method according to claim 6, characterized in that, The duration of ultraviolet irradiation is 90s to 110s.
8. The method according to claim 7, characterized in that, The ultraviolet irradiation time was 99 seconds.
9. The method according to claim 1, characterized in that, The components of the UV-curable adhesive include: isoborneol acrylate, hydroxypropyl methacrylate, acrylic acid, photoinitiator A, photoinitiator B, and silane.
10. The method according to claim 9, characterized in that, The UV-curable adhesive is UV adhesive K-306.