A device and method for determining the anatomical structure of fine roots of plants without thin sectioning

Abstract

The present application relates to the technical field of plant science, and particularly relates to a device and method for measuring the anatomical structure of plant fine roots without thin sectioning. By directly staining, stably positioning and microscopically observing the cross section of fine roots, clear and complete anatomical images of the cross section of fine roots can be obtained without dehydration, embedding and ultrathin sectioning, and reliable measurement of anatomical parameters can be realized. The steps of fixing, dehydrating, embedding and ultrathin sectioning are omitted, the sample preparation process is significantly simplified, the time from processing to imaging is shortened, the overall measurement efficiency is improved, and the device is suitable for multi-sample and batch analysis scenarios. Under the premise of ensuring the reliability of the identifiable and measurable anatomical structure, the process is simplified, the efficiency is improved and the cost is reduced, and the device has good technical value and application prospect.

Description

Technical Field

[0001] This invention relates to the field of plant science and technology, and in particular to a device and method for determining the anatomical structure of plant fine roots without the need for thin sections. Background Technology

[0002] The anatomical structure of fine roots used for plant absorption is a crucial foundation for studying the absorption and transport functions of plant roots and their functional differentiation. Measuring anatomical parameters such as root diameter, cortex thickness, stele diameter, and endodermal structure in fine root cross-sections provides vital data support for plant ecology, plant physiology, forestry, and agricultural science. Currently, research on the anatomical structure of fine roots primarily relies on microscopic anatomy techniques, the most common being paraffin sectioning combined with optical microscopy. This method typically includes sample fixation, gradient dehydration, paraffin embedding, ultrathin sectioning, staining, and microscopic imaging. By preparing root cross-sectional sections with a thickness generally less than 10 μm, observation and quantitative analysis of the internal tissue structure of fine roots can be achieved. This type of method has been widely used in relevant research, and the technical system is relatively mature.

[0003] Existing techniques are relatively mature and reliable in the study of fine root anatomy, but they still have the following shortcomings in practical applications:

[0004] ① The operation process is cumbersome and time-consuming. Existing technologies usually require multiple steps such as fixation, dehydration, embedding, sectioning, and staining. These steps are interdependent, and the overall experimental cycle is long, which is not conducive to the rapid determination of large batches of samples. This is mainly because paraffin or resin embedding and ultrathin sectioning have high requirements for sample processing conditions.

[0005] ② It is highly dependent on experimental conditions and operational techniques. Thin section preparation usually requires specialized slicing equipment and skilled operation techniques. The thickness, integrity, and surface quality of the slices all significantly affect the imaging results, thereby increasing experimental costs and technical barriers.

[0006] ③ Errors are easily introduced during sample preparation. During dehydration, embedding, and sectioning, fine root tissues may shrink, deform, or have their cut surfaces damaged, affecting the accuracy of the cross-sectional structure and measurement precision. This problem is particularly prominent when processing small-diameter, soft absorbent roots.

[0007] In summary, existing technologies still have room for improvement in terms of efficiency, cost, and applicability, and cannot simultaneously meet multiple needs such as rapid measurement, high-throughput analysis, and simplified operation. Summary of the Invention

[0008] The technical problem to be solved by the present invention is to provide a device and method for measuring the anatomical structure of plant fine roots without the need for thin sections, in order to address the shortcomings of the prior art. By directly staining, stabilizing and observing the cross-section of fine roots, clear and complete anatomical images of the cross-section of fine roots can be obtained without dehydration, embedding and ultrathin sectioning, and reliable measurement of anatomical parameters can be achieved.

[0009] This invention provides a device for determining the anatomical structure of plant fine roots without the need for thin sections, including a microscope, and further comprising:

[0010] A glass slide with a through hole in the middle is placed on the stage of a microscope.

[0011] A sample container is fixedly installed inside the through hole and is used to place the root sample to be tested.

[0012] A limiting metal wire is sleeved on the outer periphery of the root sample to be tested;

[0013] The light source is fixedly installed on the outer periphery of the through hole.

[0014] According to the plant fine root anatomy measurement device without thin sectioning provided by the present invention, the bottom surface of the glass slide is fixedly provided with a circumferential array of fixing claws near the through hole.

[0015] According to the present invention, the device for determining the anatomical structure of plant fine roots without the need for thin sections includes a sample container comprising a transparent tube and a sponge. The transparent tube is detachably disposed within a through hole, and the outer diameter of the transparent tube is smaller than the central diameter of the through hole. The sponge is wrapped around the inner wall of the transparent tube to enclose the root sample to be tested, and the inner wall of the sponge forms a positioning groove. The size of the positioning groove matches the root sample to be tested. The top surface of the sponge is lower than the top surface of the transparent tube, and the sponge is a dark-colored sponge.

[0016] According to the plant fine root anatomy measurement device without thin sectioning provided by the present invention, the limiting metal wire is slidably positioned at the corresponding position of the microscope slide. The limiting metal wire includes a straight part and a circular part. One end of the straight part is fixedly connected to the circular part. The inner diameter of the circular part is larger than the outer diameter of the root sample to be tested. The circular part is sleeved on the outer periphery of the root sample to be tested. The other end of the straight part is connected to an external fixed object. The straight part can move freely.

[0017] According to the plant fine root anatomy measurement device without thin sectioning provided by the present invention, a light source is fixedly arranged around the through hole on the upper surface of the glass slide. The light source is a ring light source, and the ring light source can be white light or yellow light.

[0018] This invention also provides a method for using the above-mentioned device for determining the anatomical structure of plant fine roots without the need for thin sections, characterized by the following steps: collecting the plant roots to be tested, fixing them in FAA solution, washing them, cutting the root segments, cutting a flat root cross-section perpendicular to the root growth direction and staining it, adding half distilled water to a transparent tube, wrapping the root segment with a sponge and placing it in the transparent tube, keeping the root cross-section facing upwards, then placing the transparent tube in the through-hole of a glass slide and fixing it with a fixing claw, then using the limiting metal wire ring part to fit around the root segment, fixing the glass slide to the microscope, adjusting the root cross-section to be horizontal by moving the ring part through the straight part, and then adjusting the brightness of the light source so that the microscope can capture a clear image.

[0019] According to the method of use provided by the present invention, the root cross-section of the root sample to be tested is higher than the top of the sponge and lower than the top of the transparent tube, and the water level of the distilled water is level with the opening of the transparent tube.

[0020] Compared with the prior art, the present invention has the following advantages:

[0021] This invention provides a device and method for determining the anatomical structure of plant fine roots without the need for thin sectioning. By directly staining, stabilizing, and observing the cross-section of fine roots under a microscope, clear and complete anatomical images of the fine root cross-section can be obtained without dehydration, embedding, or ultrathin sectioning, enabling reliable determination of anatomical parameters. This eliminates steps such as fixation, dehydration, embedding, and ultrathin sectioning, significantly simplifying the sample preparation process, shortening the time from processing to imaging, and improving overall measurement efficiency. It is suitable for multi-sample and batch analysis scenarios. While ensuring the identifiability of anatomical structures and reliable measurement, it achieves process simplification, efficiency improvement, and cost reduction, demonstrating significant technical value and application prospects. Attached Figure Description

[0022] To more clearly illustrate the technical solutions in this invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of this invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0023] Figure 1 A three-dimensional view of the apparatus for measuring the anatomical structure of plant fine roots;

[0024] Figure 2 An exploded view of a device for measuring the anatomical structure of plant fine roots;

[0025] Figure 3 A cross-sectional view of the apparatus for measuring the anatomical structure of plant fine roots;

[0026] Figure 4This is a schematic diagram of the structure of a glass slide;

[0027] Figure 5 The diagrams show a comparison of the effects of Example 2 and Comparative Example 1. The first column shows Leymus chinensis, the second column shows Vitex negundo, the third column shows Hemp arvense, the fourth column shows Clematis chinensis, and the fifth column shows Allium chinense. (a) to (o) are the effect diagrams of the Example 2, where the first row shows the complete root structure, the second row shows the epidermis and cortical parenchyma cells, and the third row shows the inner epidermal cells and stele. (p) to (ad) are the effect diagrams of the Comparative Example 1, where the fourth row shows the complete root structure, the fifth row shows the epidermis and cortical parenchyma cells, and the sixth row shows the inner epidermal cells and stele.

[0028] Figure label:

[0029] 1. Glass slide; 101. Through hole; 102. Fixing claw; 2. Sample container; 201. Transparent tube; 202. Sponge; 203. Positioning groove; 3. Limiting wire; 4. Light source. Detailed Implementation

[0030] Example 1

[0031] This embodiment provides a device for measuring the anatomical structure of plant fine roots without the need for thin sections, including: a microscope; a glass slide 1 with a through hole 101 in the middle, the glass slide 1 being placed on the stage of the microscope; a sample container 2, fixedly disposed in the through hole 101 for placing the root sample to be tested; a limiting metal wire 3, sleeved on the outer periphery of the root sample to be tested; and a light source 4, fixedly disposed on the outer periphery of the through hole 101.

[0032] As per the instruction manual Figures 1-4 As shown, a fixed claw 102 is fixedly arranged in a circumferential array on the bottom surface of the slide 1 near the through hole 101. The purpose of the fixed claw 102 is to grip the transparent tube 201.

[0033] In this embodiment, the slide 1 is made of a 76 mm × 25 mm × 1.2 mm plastic sheet, and the fixing device for the transparent tube 201 can also be made of glass, with the same dimensions as the conventional slide 1. During fabrication, a cross-shaped opening can be cut at the center of the slide 1 using a sharp blade, and the material at the opening can be turned outwards to form several fixing claws 102. Alternatively, slightly inwardly tilted metal or other suitable material springs can be installed at the corresponding positions of the fixing claws 102 in the slide 1. The core purpose is to achieve a stable grip on the transparent tube 201. The opening needs to be machined into a circular through hole 101. The main function of the fixing claws 102 is to keep the transparent tube 201 vertical, facilitating subsequent microscopic observation.

[0034] In practical applications, slide 1 is composed of two overlapping plastic plates. The upper plastic plate has a regular octagon shaped at its center. The distance between the two parallel sides of this octagon should be slightly larger than the outer diameter of the transparent tube 201. This ensures the transparent tube 201 can be inserted smoothly while preventing significant misalignment during placement. During fabrication, first, use a blade to make marks along the eight sides of the octagon, being careful not to cut through the plastic plate. Then, cut along the diagonal of the octagon, turning the cut portion outwards to form eight retaining claws 102. These retaining claws 102 naturally form an inward elasticity, stably clamping the transparent tube 201 and ensuring its stability during placement.

[0035] Figure 1 The fixing claw 102 shown is a circumferentially arranged metal spring, but in actual use, the method of overlapping and drilling holes in two plastic plates is simpler and more applicable. Figure 4 As shown.

[0036] As for the other plastic plate located below, it serves two purposes: first, it provides reinforcement because the plastic plate itself is relatively soft; second, it restricts the outward rotation of the fixing claw 102. The opening position of the second plate is the same as that of the first plate, but the opening shape is a square. The side length is slightly smaller than the distance between the two parallel sides of the regular octagon of the first plate, but larger than the outer diameter of the transparent tube 201, ensuring that the transparent tube 201 can be inserted. It also provides an inward constraint force to the fixing claw 102, making it easier to clamp the transparent tube 201.

[0037] Through testing, it was found that when using fixing claws 102 made of different materials, there is no need to pursue excessively high clamping stability. This is because in actual use, after the slide 1 is placed on the microscope stage, the microscope itself has a fixing structure for the slide 1, which can effectively limit the up, down, left, and right movement of the slide 1 and ensure its placement stability. Therefore, there is no need for the fixing claws 102 to provide extremely strong clamping force.

[0038] The slide 1 and the fixing claw 102 can be integrally molded or manufactured separately and then assembled, depending on the actual needs. Both the fixing claw 102 and the transparent tube 201 are made of plastic, but the fixing claw 102 is made of a softer material and will not scratch the transparent tube 201. If other fixing claw 102 implementations are used, such as installing slightly inward-tilted metal or other suitable material springs, although scratches may occur on the side wall of the transparent tube 201, these scratches will not affect the normal use of the transparent tube 201 or subsequent observation, because microscope observation is performed from above, and side wall scratches do not affect the field of view. Furthermore, the transparent tube 201 can be modified from a common 2 mL centrifuge tube in the laboratory. This type of consumable is inexpensive and readily available, effectively reducing experimental costs.

[0039] The sample container 2 includes a transparent tube 201 and a sponge 202. The transparent tube 201 is detachably installed inside the through hole 101. The outer diameter of the transparent tube 201 is slightly smaller than the central diameter of the through hole 101. The sponge 202 is wrapped around the inner wall of the transparent tube 201 to wrap the root sample to be tested. The inner wall of the sponge 202 forms a positioning groove 203. The size of the positioning groove 203 matches the root sample to be tested. The top surface of the sponge 202 is lower than the top surface of the transparent tube 201. The sponge 202 is a dark-colored sponge 202.

[0040] In this embodiment, the transparent tube 201 is a 2 mL transparent plastic centrifuge tube with an outer diameter of 10.4 mm, a wall thickness of 0.8 mm, and a depth of 20 mm. The sponge 202 is a dark-colored absorbent sponge, which provides a dark background, thus improving the observation of the root outline. The sponge 202 measures 20 mm × 10 mm × 4 mm and is fully soaked with water before use. The transparent tube 201 is made by cutting a 2 mL centrifuge tube, with a height of approximately 2 cm. Its height is not strictly required; it only needs to be 1 cm higher than the sponge 202 to ensure that the sponge 202 can be easily inserted into the tube. The sponge 202 is elastic. After the sponge 202 is wrapped around the inner wall of the transparent tube 201, it forms a positioning groove 203. The inner diameter of the positioning groove 203 matches the root sample to be tested and is used to place and limit the root sample to be tested. After the sponge 202 wraps the root sample to be tested, its cross-sectional area will be larger than that of the transparent tube 201. After it is inserted into the transparent tube 201, the sponge 202 will expand outward, thereby fixing it inside the tube and effectively preventing the transparent tube 201 from floating up and down.

[0041] Sample container 2 is a detachable and reusable unit, which facilitates the replacement of the root sample to be tested.

[0042] The limiting metal wire 3 is slidably set at the corresponding position of the microscope slide 1. The limiting metal wire 3 includes a straight part and a circular part. One end of the straight part is fixedly connected to the circular part. The inner diameter of the circular part is larger than the outer diameter of the root sample to be tested. The circular part is sleeved on the outer periphery of the root sample to be tested. The other end of the straight part is connected to an external fixed object. The straight part can move freely.

[0043] In this embodiment, the limiting metal wire 3 is made of 1.2 mm diameter aluminum wire. Various types of metal wire with good plasticity can be used for the limiting metal wire 3. Its circular part is fitted around the outer periphery of the root sample to be tested, and the other end of the straight part can be fixed to a counterweight with a certain weight that is not easily moved. Alternatively, the limiting metal wire 3 can be directly attached to or pasted onto the microscope. It is necessary to ensure that the limiting metal wire 3 has a section that can move freely. Its core function is to move the root cross-section of the root sample to a horizontal plane or near a horizontal plane to meet the needs of subsequent observation.

[0044] A light source 4 is fixedly installed around the through hole 101 on the upper surface of the glass slide 1. The light source 4 is a ring light source, and the ring light source can be white light or yellow light.

[0045] In this embodiment, the light source 4 is a yellow or white light strip with a rated voltage of 5 V and a diameter of 1 mm that emits light around the entire circumference. It is combined with a plastic aluminum wire with a diameter of 2 mm bent into a ring shape. The light source 4 is set horizontally, with its inner diameter slightly larger than the outer diameter of the transparent tube 201. The light-emitting surface is slightly higher than or level with the plane of the transparent tube 201. The light strip of the light source 4 is selected as a light strip with its own battery. When the light strip runs out of power, a new light strip can be replaced.

[0046] Example 2

[0047] This embodiment provides a method for using the device provided in Embodiment 1.

[0048] ① Device fabrication:

[0049] The slide 1 is made of plastic and measures 76 mm × 25 mm × 1.2 mm. A cross-shaped opening is made in the center using a sharp blade and then turned outward to form a fixing claw 102. The size of the through hole 101 is 10.5 mm. The inner diameter of the through hole 101 is slightly larger than the outer diameter of the transparent tube 201 (10.4 mm) to ensure that the transparent tube 201 can be inserted smoothly. At the same time, the difference in distance should preferably not exceed 1 mm to avoid the transparent tube 201 from tilting after placement due to excessive difference. Even with the fixing claw 102 holding it, it is difficult to avoid tilting.

[0050] Transparent tube 201 is made by horizontally cutting a 20 mm high section from a 2 mL transparent plastic centrifuge tube. The centrifuge tube has an outer diameter of 10.4 mm and a wall thickness of 0.8 mm. Sponge 202 has dimensions of 20 mm × 10 mm × 4 mm and is fully soaked in water before use.

[0051] The limiting metal wire 3 is made of aluminum wire with a diameter of 1.2 mm. The height of the straight part of the limiting metal wire 3 must meet specific requirements: it must be higher than the height from the upper surface of the sponge 202 to the opening of the transparent tube 201, so that its circular part can be inserted into the transparent tube 201 to move the root sample to be tested; at the same time, this height must be less than the distance from the upper surface of the sponge 202 to the microscope objective lens to avoid hindering the switching operation of the objective lens. Since the straight part of the limiting metal wire 3 is malleable, the above height requirements are very easy to achieve.

[0052] Light source 4 is a yellow light strip with a rated voltage of 5 V and a diameter of 1 mm that emits light around the entire circumference. It is combined with a 2 mm diameter plastic aluminum wire bent into a ring shape.

[0053] ② Preparation of staining agent: Prepare a 0.05% toluidine blue solution using distilled water.

[0054] ③ Sample preparation: Root systems of five plants were collected from the grassland plant cultivation resource nursery of the Jilin Songnen Grassland Ecosystem National Field Scientific Observation and Research Station of Northeast Normal University. The plants were: Vitex multistem, Fern root, Clematis chinensis, Wild onion, and Sheepgrass.

[0055] These five plants belong to different families, and their roots vary significantly in thickness and hardness, making them representative. Based on morphological grading standards, primary absorbing roots of each species were selected, collected, and fixed in FAA solution (formalin-acetic acid-ethanol mixture).

[0056] ④ Experimental procedure: Remove the root sample from the FAA fixative, wash it with distilled water, and cut a root segment of about 10 mm in length.

[0057] Cut a flat cross-section of the root perpendicular to the root growth direction with a sharp blade to serve as the observation surface. Stain the root cross-section in 0.05% toluidine blue solution for about 1 second, then wash away excess dye with water. The resulting root segment is the root sample to be tested.

[0058] Wrap the root sample to be tested in a fully absorbent sponge 202, exposing approximately 1 mm of the stained root cross-section. Pre-fill a transparent tube 201 with half its volume of water. Place the sponge 202 containing the root sample into the tube, keeping the root cross-section facing upwards and slightly below the water level (within 1 mm) to avoid water contamination of the lens when switching to high magnification. The water level should be approximately level with or slightly lower than the tube opening. Thus, the transparent tube 201 and sponge 202 together form sample container 2.

[0059] Place it on glass slide 1, ensuring the plane of the tube opening is 1–2 mm above the surface of slide 1. Fix slide 1 to the microscope stage and install light source 4, ensuring its luminous surface is level with or slightly higher than the cross-section of the root sample. Use the circular part of the limiting wire 3 to cover the exposed end of the root sample, and adjust the root cross-section to be horizontal using the straight part.

[0060] Adjust the brightness of the light source under the microscope and the light source 4 set in this device to find a clear field of view and take pictures. Since it is difficult to ensure that the root cross-section is completely flat by hand, especially when the root diameter is large, different areas may be on different focal planes. Therefore, clear images can be taken for each area separately, and then image processing software can be used to perform depth-of-field synthesis through image stacking to finally obtain a clear and complete image of the entire field of view.

[0061] Comparative Example 1

[0062] This comparative example provides a traditional paraffin sectioning method.

[0063] S1. Sample preparation is the same as in Example 2;

[0064] S2. Dehydration: The root segments are sequentially placed in distilled water, 30% ethanol, 50% ethanol, 70% ethanol, 80% ethanol, 95% ethanol, and anhydrous ethanol (twice) for gradient dehydration;

[0065] S3, Transparency: The dehydrated root segment samples were placed in anhydrous ethanol-xylene mixed solutions with volume ratios of 2:1, 1:1, and 1:2, respectively, and then treated twice with pure xylene to gradually replace the ethanol.

[0066] S4. Wax impregnation: The transparent root segment samples are impregnated successively in a 1:1 xylene-paraffin mixture and twice in pure paraffin.

[0067] S5. Embedding: Embed the root segment sample with liquid pure paraffin and allow it to cool naturally;

[0068] S6. Slicing and Slide Preparation: Slicing, gluing, spreading and baking the slides;

[0069] S7. Dewaxing: The sections were placed in pure xylene, an anhydrous ethanol-xylene mixed solution (1:1 volume ratio), anhydrous ethanol, 95% ethanol, 80% ethanol and 70% ethanol in sequence to gradually dewax and hydrate.

[0070] S8. Staining: Double staining with safranin-fast green was performed, and the slides were cleaned and then mounted for observation.

[0071] The comparative method is compared with the method in Embodiment 2 of this application, and the following table is obtained:

[0072] Table 1

[0073]

[0074] The absorption fine root cross-section microscopic imaging method established in Example 2, which does not require thin sections, can obtain clear and complete anatomical images of fine root cross sections by directly staining, stabilizing and observing the fine root cross sections without dehydration, embedding and ultrathin sectioning, and can reliably measure anatomical parameters.

[0075] Experimental results are as follows Figure 5 As shown, the results indicate that in the root cross-sectional images obtained using this method, the main structures such as root epidermal cells, cortical parenchyma cells, endodermis, and stele are clearly distinguishable, and the U-shaped thickening feature of the endodermal cell wall is also clearly visible. The endodermis is an important structure for distinguishing the stele from the cortex in absorptive roots.

[0076] Image comparison with traditional paraffin sectioning shows that the present invention is consistent with it in terms of measurability and structural identification of key anatomical parameters such as root diameter, cortex thickness and stele diameter, and can meet the technical requirements for observation and quantitative analysis of fine root anatomical structures.

[0077] This invention eliminates steps such as fixation, dehydration, embedding, and ultrathin sectioning, significantly simplifying the sample preparation process, shortening the time from processing to imaging, and improving the overall measurement efficiency. It is suitable for multi-sample and batch analysis scenarios.

[0078] Actual measurements show that the traditional paraffin sectioning method requires approximately 29.3 hours from dehydration to imaging, while the method provided by this invention can achieve similar imaging results within 17 minutes. Furthermore, this method does not rely on specialized sample preparation equipment such as microtome machines, has lower operational skill requirements, can be carried out under standard laboratory conditions, and possesses good operability and potential for widespread application.

[0079] Furthermore, by reducing the chemical and mechanical treatment of fine root tissues, this invention lowers the risk of shrinkage, deformation, or breakage that may occur during dehydration, embedding, and sectioning. This helps maintain the relative authenticity of the root cross-sectional structure and, theoretically, improves the stability and repeatability of measurement results. In summary, this invention simplifies the process, improves efficiency, and reduces costs while ensuring anatomical structure identification and reliable measurement, demonstrating significant technical value and application prospects.

[0080] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention in any way. Any simple modifications, alterations, and equivalent changes made to the above embodiments based on the inventive essence shall still fall within the protection scope of the present invention.

Claims

1. A device for determining the anatomical structure of plant fine roots without the need for thin sections, comprising a microscope, characterized in that, Also includes: A glass slide with a through hole in the middle is placed on the stage of a microscope. A sample container is fixedly installed inside the through hole and is used to place the root sample to be tested. A limiting metal wire is sleeved on the outer periphery of the root sample to be tested; The light source is fixedly installed on the outer periphery of the through hole.

2. The device for determining the anatomical structure of plant fine roots without the need for thin sections according to claim 1, characterized in that, The bottom surface of the glass slide is fixedly provided with a circumferential array of fixing claws near the through hole.

3. The device for determining the anatomical structure of plant fine roots without the need for thin sections according to claim 1, characterized in that, The sample container includes a transparent tube and a sponge. The transparent tube is detachably installed inside the through hole. The outer diameter of the transparent tube is smaller than the central diameter of the through hole. The sponge is wrapped around the inner wall of the transparent tube to wrap the root sample to be tested. The inner wall of the sponge forms a positioning groove. The size of the positioning groove matches the root sample to be tested. The top surface of the sponge is lower than the top surface of the transparent tube. The sponge is a dark-colored sponge.

4. The device for determining the anatomical structure of plant fine roots without the need for thin sections according to claim 3, characterized in that, The limiting metal wire is slidably set at the corresponding position of the microscope slide. The limiting metal wire includes a straight part and a circular part. One end of the straight part is fixedly connected to the circular part. The inner diameter of the circular part is larger than the outer diameter of the root sample to be tested. The circular part is sleeved on the outer periphery of the root sample to be tested. The other end of the straight part is connected to an external fixed object. The straight part can move freely.

5. The device for determining the anatomical structure of plant fine roots without the need for thin sections according to claim 1, characterized in that, A light source is fixedly arranged around the through hole on the upper surface of the glass slide. The light source is a ring light source, and the ring light source can be white light or yellow light.

6. A method of using the plant fine root anatomy measurement device without thin sectioning as described in any one of claims 1-5, characterized in that, Includes the following steps: The plant roots to be tested are collected, fixed in FAA solution, washed, and root segments are cut. A flat root cross-section is cut perpendicular to the root growth direction and stained. Half distilled water is added to a transparent tube, and the root segment is wrapped with a sponge and placed in the transparent tube, keeping the root cross-section facing upward. The transparent tube is then placed in the through-hole of a glass slide and fixed with a fixing claw. Then, the circular part of the limiting metal wire is placed around the root segment. After fixing the glass slide to the microscope, the root cross-section is adjusted to be horizontal by moving the circular part through the straight part. By adjusting the brightness of the light source, the microscope can capture a clear image.

7. The method of use according to claim 6, characterized in that, The root cross-section of the root sample to be tested is higher than the top of the sponge and lower than the top of the transparent tube, and the level of the distilled water is level with the opening of the transparent tube.

Images