In-situ layered sediment sampling and culturing apparatus and method

Abstract

This invention provides an in-situ sediment stratification sampling and culture device and method, including a base with sample chambers arranged circumferentially on the base and a turret at the center of the base. A sediment sampling and dispensing device is mounted on the turret. The sampling and dispensing device includes a sediment sampling tube, a tube extension mechanism, a pushing mechanism, and a stratification and dispensing mechanism. The tube extension mechanism drives the sampling tube to insert into the sediment for sampling; the pushing mechanism pushes the sediment out of the tube opening; and the stratification and dispensing mechanism cuts and stratifies the ejected sediment. The turret drives the sampling and dispensing device to rotate, sequentially dispensing the stratified samples into each sample chamber. Each sample chamber contains a fixative for bio-information fixation of the sediment. This invention achieves integrated in-situ low-disturbance sediment sampling, vertical stratification cutting, automated dispensing, and sample fixation, effectively avoiding information loss during sample transfer.

Description

Technical Field

[0001] This invention relates to the field of marine biological sampling technology, specifically to an in-situ stratified sampling and culture device and method for sediments. Background Technology

[0002] Seafloor sediments cover almost the entire seabed surface, playing a crucial role in recording historical environmental information, providing habitats for organisms, and supporting the storage and transformation of materials. First, organic matter and other nutrients in sediments are formed in seawater, slowly accumulating on the seabed through sedimentation, thus preserving historical information about the sedimentary environment. The sedimentation rate in deep-sea plains is only about centimeters per millennium, allowing for the reconstruction of historical environments through stratified sediment analysis. Second, surface sediments are more disturbed by bottom currents and benthic organisms, while material transfer in deep sediments is primarily through diffusion, exhibiting a significant chemical gradient. Especially in special geological environments such as deep-sea cold seeps and hydrothermal vents, the vertical chemical gradient changes dramatically, rapidly transitioning from an oxygen-rich layer to an anoxic layer with nitrate, sulfate, and iron as electron acceptors within a few centimeters. Third, the estimated total number of cells in the sediments is 2.9 × 10²⁹, exceeding the microbial cell count in ocean water. Microbial content decreases rapidly with decreasing organic matter content in the sediments, showing significant vertical differences in cell quantity and community structure. Therefore, in-situ vertical stratification sampling and preservation of sediments is a common need for multiple disciplines such as marine geology, chemistry, and biology.

[0003] Traditional methods for sediment stratification studies involve collecting sediment cores (such as via tubing or gravity columns) onto a deck for further stratification. This process is fraught with challenges due to sample oscillations during ascent, temperature fluctuations, and decompression, disturbing the sediment stratification, particularly affecting surface sediments and leading to the loss of environmental parameters and sample information. Furthermore, sediment sampling, distribution, transfer, and preservation all rely on mechanical methods, resulting in numerous, complex, and unreliable steps. Internationally, there has been a lack of in-situ stratification sampling and cultivation equipment specifically designed for deep-sea sediments. Summary of the Invention

[0004] To address the shortcomings of existing technologies, the purpose of this invention is to provide an in-situ sediment stratification sampling and cultivation device and method.

[0005] An in-situ sediment stratification sampling and culture device according to the present invention includes: Equipment base; Multiple sample chambers are arranged circumferentially on the base of the device; A turret is located at the center of the equipment base; A sediment cannula sampling and dispensing device is mounted on the turret and can rotate with the turret to be aligned with any of the sample chambers; The sediment cannula sampling and dispensing device includes: Sediment sampling cannula, used for inserting into sediments to collect samples; The cannula telescopic mechanism drives the sediment sampling cannula to be inserted into or withdrawn from the sediment; A sediment sample pushing mechanism is used to push the sediment sample out of the sediment sampling tube; A sediment stratification and sampling mechanism is installed at the port of the sediment sampling tube and is used to cut and stratify the ejected sediment. The sample chamber contains a fixative for fixing the biological information of the stratified sediment samples.

[0006] Preferably, the turret is further provided with a pitch drive mechanism for driving the sediment insertion tube sampling and dispensing device to switch between a vertical sampling position and a horizontal dispensing position.

[0007] Preferably, the sample chamber includes: Cultivation chamber; The culture chamber cover is rotatably connected to the top of the culture chamber body and is kept closed by a torsion spring. The closing trigger lever is used to lock the culture chamber cover in the open state and to automatically close the culture chamber cover after release.

[0008] Preferably, the sample chamber further includes: A culture substrate tube is disposed on the upper part of the culture chamber, contains culture substrate, and is sealed by a culture substrate tube membrane; A substrate release and mixing actuator is rotatably mounted on the culture chamber and is used to pull and rupture the culture substrate membrane during rotation to release the culture substrate.

[0009] Preferably, the sample chamber further includes: A fixative chamber is located at the bottom of the culture chamber, contains fixative, and is sealed by a fixative chamber membrane; A fixative release and mixing actuator is rotatably mounted on the culture chamber and is used to pull and tear the fixative chamber membrane during rotation to release the fixative.

[0010] Preferably, the sediment cannula sampling and dispensing device further includes: A sample chamber cap sealing trigger and substrate release mixing mechanism is used to trigger the closing of the sample chamber and the release of the culture substrate; A fixative release and mixing drive mechanism is used to trigger the release of the fixative from the sample chamber.

[0011] A method for in-situ stratified sampling and cultivation of sediments according to the present invention includes the following steps: Step S1: Deploy the equipment at the seabed sediment interface; Step S2: The sediment sampling cannula is inserted into the sediment by driving the cannula telescopic mechanism to complete the sampling; Step S3: Rotate the sediment cannula sampling dispenser to the top of the target sample chamber using a turret; Step S4: Push the sediment sample out of the insertion tube using the sediment sample pushing mechanism; Step S5: During the push-out process, the sediment is layered and cut by the sediment stratification and sampling mechanism, so that the layered samples fall into the sample chamber; Step S6: Trigger the sample chamber lid to close and release the fixative to fix the bio-information of the sample.

[0012] Preferably, before inserting the sediment sampling tube into the sediment, the method further includes switching the sediment tube sampling and dispensing device to a vertical position via a pitch drive mechanism; after sampling is completed, it is switched to a horizontal position for dispensing.

[0013] Preferably, after the sample chamber is closed, the method further includes: releasing the culture substrate and mixing it with the sediment sample for in-situ culture; and releasing a fixative for bio-information fixation after the culture is completed.

[0014] Preferably, the sediment sampling and dispensing device is rotated by a turret, and the sampling steps are repeated in sequence with multiple sample chambers to achieve layered dispensing and fixation of multiple sediment samples.

[0015] Compared with the prior art, the present invention has the following beneficial effects: 1. This invention achieves low-disturbance positioning of the sediment-water interface by gently lowering a large-area chassis to the bottom. After sampling, vertical stratification and fixation are performed directly in situ, avoiding temperature rise, pressure drop, and vibration disturbance during sample transfer, thus preserving the original structure and biological information of the sediment to the greatest extent.

[0016] 2. This invention integrates functions such as cannulation sampling, rotating turret, quantitative pushing, layered cutting, rotary dispensing, automatic capping, and reagent release, transforming complex operations into an automated process. This significantly reduces manual intervention, improves the reliability and efficiency of deep-sea operations, and meets the sample allocation needs of multiple disciplines.

[0017] 3. In this invention, the sample chamber contains culture substrate and fixative, which can be released and mixed through mechanical linkage to achieve in-situ culture and fixation of biological information. Simultaneously, it is equipped with an electrochemical sensor to collect vertical profile chemical parameters, supporting multiple parallel experiments and enhancing the comprehensive capabilities of deep-sea environment research. Attached Figure Description

[0018] Other features, objects, and advantages of the present invention will become more apparent from the following detailed description of non-limiting embodiments with reference to the accompanying drawings: Figure 1 This is a schematic diagram of the system composition of the sediment stratification sampling and fixation equipment disclosed in this invention; Figure 2 This is a diagram showing the composition of the sediment cannulation sampling and dispensing device disclosed in this invention; Figure 3 This is a structural diagram of the sample chamber disclosed in this invention; Figure 4 This is a diagram illustrating the working principle and breakdown of the sediment stratification sampling and fixation equipment disclosed in this invention.

[0019] Explanation of reference numerals in the attached figures: Detailed Implementation

[0020] The present invention will now be described in detail with reference to specific embodiments. These embodiments will help those skilled in the art to further understand the present invention, but do not limit the invention in any way. It should be noted that those skilled in the art can make several changes and improvements without departing from the concept of the present invention. These all fall within the protection scope of the present invention.

[0021] The in-situ sediment stratification sampling and culture device disclosed in this invention mainly consists of a sediment cannula sampling and dispensing device 1, a sample chamber 2, and a device base 3, as described above. Figure 1 As shown, the sample chambers 2 are arranged in a circular pattern on the equipment base 3. A turret is installed on the equipment base 3, which can carry the sediment cannula sampling and dispensing device 1. The turret rotates around the center of the equipment base 3 to realize the operation of transferring, sealing, culturing, and fixing samples to each sample chamber 2. The turret is also equipped with a pitch drive mechanism, which can drive the sediment cannula sampling and dispensing device 1 to switch between horizontal and vertical positions.

[0022] Sediment cannulation sampling divider consists of Figure 2 The mechanism shown includes a sediment sampling tube 11, a sediment stratification and sampling mechanism 12, a tube extension mechanism 13, a sediment sample pushing mechanism 14, a sample chamber cap triggering and substrate release and mixing mechanism 15, and a fixative release and mixing drive mechanism 16.

[0023] The sediment sampling tube 11 serves as a sampler for seabed sediments. A tube extension mechanism 13 drives the sediment sampling tube 11 in reciprocating motion for sampling. A sediment sample pushing structure 14 is located at the tail end of the sediment sampling tube 11. After sampling, the pushing structure 14 extends into the sediment sampling tube 11 to push the seabed sediments out, transferring the marine sediments to the sample chamber 2. A sediment stratification and separation mechanism 12 is located at the head end of the sediment sampling tube 11. During the transfer of marine sediments, the sediment stratification and separation mechanism 12, upon activation, cuts off the marine sediments through its own rotation and blocks the outlet of the sediment sampling tube 11.

[0024] The working method of the sediment cannula sampling and sampler 11 is as follows: the cannula telescopic mechanism 13 drives the sediment sampling cannula 11 to accurately insert into the sediment at the set depth to collect sediment samples. After sampling, the cannula telescopic mechanism 13 drives the sediment sampling cannula to retract. The sediment sample pushing mechanism 14 can push out the collected sediment according to the designed length, and the sediment layering and sample separation mechanism 12 set at the cannula opening can cut and separate the samples into layers. The sample chamber cap triggering and substrate release and mixing mechanism 15 can operate the sample chamber 2 to achieve cap sealing. The fixative release and mixing drive mechanism 16 can realize the release and mixing of fixative in the chamber to achieve biological information fixation of the cultured sample.

[0025] The sample chamber structure consists of, for example, Figure 3 The sample chamber includes a culture chamber cover 21, a closing trigger rod 22, a culture chamber body 23, a culture substrate tube 24, a substrate release and mixing actuator 25, a fixative release and mixing actuator 26, a fixative chamber membrane 27, and a culture substrate tube membrane 28. The culture chamber cover 21 is located on top of the culture chamber body 23 and is rotatably connected to it. A torsion spring is installed at the connection between the culture chamber cover 21 and the culture chamber body 23. Under the elastic force of the torsion spring, the culture chamber cover 21 covers and presses against the opening of the culture chamber body 23. The culture chamber cover 21 includes an operating rod with a U-shaped opening at one end. When the closing trigger rod 22 engages the U-shaped opening, the culture chamber cover 21 remains open. When the closing trigger rod 22 is rotated to disengage from the U-shaped opening, the culture chamber cover 21 remains closed under the elastic force of the torsion spring. A handle is provided on the closing trigger rod 22. The sample chamber lid can be opened or closed by rotating the handle of the closing trigger rod 22 clockwise or counterclockwise.

[0026] The upper part of the culture chamber 23 has a substrate tube 24, which isolates the substrate from the external environment through a substrate membrane 28. The substrate is pre-prepared and placed into the substrate tube 24 before deployment. A substrate release and mixing actuator 25 is installed on the culture chamber 23, penetrating and rotatably connected to it. A handle is located at the end of the actuator outside the culture chamber 23, and a puller is located at the end inside the chamber. A traction rope is installed on the substrate membrane 28, which can be nested on the puller. When the sample chamber cap is triggered and the substrate release and mixing mechanism 15 rotates, the handle of the substrate release and mixing actuator 25 rotates, causing the puller to rotate. The traction rope wraps around the puller, pulling the substrate membrane 28 out and releasing the substrate.

[0027] The fixative chamber at the bottom of the culture chamber 23 is pre-filled with fixative, which is isolated from the external environment by the fixative membrane 27 in the middle of the culture chamber. A fixative release and mixing actuator 26 is provided on the culture chamber 23. The structure of the fixative release and mixing actuator 26 is similar to that of the substrate release and mixing actuator 25. The fixative chamber membrane 27 is also provided with a traction rope. The fixative release and mixing drive mechanism 16 actuates the handle of the fixative release and mixing actuator 26, thereby driving the traction device of the fixative release and mixing actuator 26 to rotate. The traction rope rotates around the traction device, thereby pulling the fixative chamber membrane 27.

[0028] After the sediment samples are loaded, the sample chamber capping trigger and substrate release and mixing mechanism 15 triggers the cap closing trigger rod 22, closing the sample chamber 2 and isolating it from the external environment. The substrate is released by rotating the substrate release and mixing actuator 25, and the substrate and sample are mixed and stirred to begin culturing. After culturing, the fixative is released by rotating the fixative release and mixing actuator 26 and mixed and stirred to complete the fixation of biological information in the cultured sample.

[0029] Equipment workflow as follows Figure 4 As shown. After the equipment is positioned on the seabed, the insertion tube is first erected and pushed into the sediment for sampling. After sampling, the insertion tube is switched to a horizontal position. The insertion tube end is rotated to the target sample chamber, and the sediment sample is quantitatively dispensed. The sample distribution mechanism distributes the sample into the sample chamber, the sample chamber cover is closed, and the fixative is released to fix the sample. By switching the angle of the insertion tube, this step is repeated to start the sampling and experiment for all 10 sample chambers. Ten fixation experiments are then completed to achieve biological information fixation.

[0030] Due to the influence of high-pressure seawater, underwater actuators experience significant operating resistance, frequently resulting in incomplete or even non-functional actions. This project proposes to utilize high-power-density servo motors and controllers, coupled with high-precision, high-torque robot joint reducers, to enhance the accuracy and reliability of underwater actions. Simultaneously, high-precision angle feedback sensors will be installed to monitor the actuator's operation and accuracy.

[0031] In the description of this application, it should be understood that the terms "upper", "lower", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are only for the convenience of describing this application and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on this application.

[0032] Specific embodiments of the present invention have been described above. It should be understood that the present invention is not limited to the specific embodiments described above, and those skilled in the art can make various changes or modifications within the scope of the claims, which do not affect the essence of the present invention. Unless otherwise specified, the embodiments and features described in this application can be arbitrarily combined with each other.

Claims

1. A sediment in-situ stratified sampling and culture device, characterized in that, include: Equipment base (3); Multiple sample chambers (2) are arranged circumferentially on the equipment base (3); A turret is located at the center of the equipment base (3); A sediment cannula sampling device (1) is installed on the turret and can rotate with the turret to align with any of the sample chambers (2). The sediment cannula sampling and dispensing device (1) includes: Sediment sampling cannula (11) is used for insertion into sediments for sampling; The cannula telescopic mechanism (13) drives the sediment sampling cannula (11) to be inserted into or withdrawn from the sediment; A sediment sample pushing mechanism (14) is used to push the sediment sample out of the sediment sampling tube (11); A sediment stratification and sampling mechanism (12) is installed at the port of the sediment sampling tube (11) and is used to cut and stratify the ejected sediment. The sample chamber (2) contains a fixative for fixing the biological information of the stratified sediment samples.

2. The in-situ sediment stratification sampling and cultivation equipment according to claim 1, characterized in that, The turret is also equipped with a pitch drive mechanism, which is used to drive the sediment insertion tube sampling and dispensing device (1) to switch between the vertical sampling position and the horizontal dispensing position.

3. The in-situ sediment stratification sampling and cultivation equipment according to claim 1, characterized in that, The sample chamber (2) includes: Culture chamber (23); The culture chamber cover (21) is rotatably connected to the top of the culture chamber body (23) and is kept closed by a torsion spring; The closing trigger rod (22) is used to lock the culture chamber cover (21) in the open state and to automatically close the culture chamber cover (21) after release.

4. The in-situ sediment stratification sampling and culture equipment according to claim 3, characterized in that, The sample chamber (2) also includes: A culture substrate tube (24) is disposed on the upper part of the culture chamber (23), contains culture substrate, and is sealed by a culture substrate tube membrane (28); The substrate release and mixing actuator (25) is rotatably mounted on the culture chamber (23) and is used to pull and rupture the culture substrate membrane (28) during rotation to release the culture substrate.

5. The in-situ sediment stratification sampling and cultivation equipment according to claim 3, characterized in that, The sample chamber (2) also includes: A fixative chamber is located at the bottom of the culture chamber (23), contains fixative, and is sealed by a fixative chamber membrane (27); A fixative release and mixing actuator (26) is rotatably mounted on the culture chamber (23) and is used to pull and tear the fixative chamber membrane (27) during rotation to release the fixative.

6. The in-situ sediment stratification sampling and cultivation equipment according to claim 1, characterized in that, The sediment cannulation sampling divider (1) also includes: The sample chamber sealing trigger and substrate release mixing mechanism (15) is used to trigger the closing of the sample chamber (2) and the release of the culture substrate; A fixative release and mixing drive mechanism (16) is used to trigger the release of fixative from the sample chamber (2).

7. A method for in-situ stratified sampling and cultivation of sediments using the equipment described in any one of claims 1 to 6, characterized in that, Includes the following steps: Step S1: Deploy the equipment at the seabed sediment interface; Step S2: Drive the sediment sampling cannula (11) into the sediment through the cannula telescopic mechanism (13) to complete the sampling; Step S3: Rotate the sediment cannula sampling sampler (1) above the target sample chamber (2) using a turret; Step S4: Push the sediment sample out of the insertion tube using the sediment sample pushing mechanism (14); Step S5: During the push-out process, the sediment is layered and cut by the sediment layering and sampling mechanism (12) so that the layered samples fall into the sample chamber (2); Step S6: Trigger the closing of the sample chamber (2) and release the fixative to fix the bio-information of the sample.

8. The method for in-situ stratified sampling and cultivation of sediments according to claim 7, characterized in that, Before the sediment sampling cannula (11) is inserted into the sediment, the sediment sampling divider (1) is switched to a vertical position by a pitch drive mechanism; after sampling is completed, it is switched to a horizontal position for the separation operation.

9. The method for in-situ stratified sampling and cultivation of sediments according to claim 7, characterized in that, After the sample chamber (2) is closed, the process also includes: releasing the culture substrate and mixing it with the sediment sample for in-situ culture; and releasing the fixative for biological information fixation after the culture is completed.

10. The method for in-situ stratified sampling and cultivation of sediments according to claim 7, characterized in that, By rotating the sediment insertion tube sampling and dispensing device (1) by turret, the sampling steps are repeated by sequentially aligning it with multiple sample chambers (2) to achieve layered dispensing and fixation of multiple sediment samples.

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