Micro-tissue sample on-line eddy current sampling device

Abstract

The invention relates to a micro-tissue sample online vortex sampling device which comprises a cutting device, a sample placing device and a vortex sampling device, the sample placing device is used for placing an original micro-tissue sample, and a hole with a preset aperture is formed in the sample placing device; the cutting device is used for cutting an original micro-tissue sample arranged on the sample placing device, and the vortex sampling device is arranged below the sample placing device and is used for collecting the cut micro-tissue sample released from an opening of the sample placing device in a vortex sampling manner. According to the invention, on-line extraction of a sample obtained by laser microdissection is realized, sampling in a flow process is realized by means of surface tension in a liquid vortex manner, and sample transmission is realized by means of a liquid injection pump, so that subsequent analysis of the micro-tissue sample is facilitated.

Description

Technical Field

[0001] The present invention relates to the technical field of micro-tissue online sampling, and in particular to an online eddy current sampling device for micro-tissue samples. Background Art

[0002] The tissue in an organism is a complex environment, containing several or even dozens of cells. In addition, it also contains a large amount of intercellular matrix. In recent years, with the continuous improvement of gene sequencing technology and the rapid development of personalized medicine, single-cell analysis has become the forefront and top priority of cell and life science research. Therefore, the establishment of accurate single-cell analysis methods will greatly promote cell biology research and be widely used in the field of life sciences.

[0003] Cell capture and single-cell extraction are prerequisites for single-cell analysis. Currently, single-cell characterization methods are mainly limited to two aspects. The first is sampling difficulties, lack of online flow sampling of microtissue particle samples, and the second is the separation of the front-end sampling and back-end management processes. The continuity from sampling to transmission is insufficient, resulting in the sampled cells being unable to be directly and smoothly connected to the subsequent processes.

[0004] Currently, there is a lack of an effective online sampling method for samples cut by laser microscopy. Since the microtissue samples after laser microscopy have a certain mechanical toughness, they will adhere to the slices and are not easy to fall off. Summary of the invention

[0005] In response to the above-mentioned technical problems, an online eddy current sampling device for micro-tissue samples is provided.

[0006] The technical means adopted by the present invention are as follows:

[0007] A microtissue sample online eddy current sampling device comprises a cutting device, a sample placement device and an eddy current sampling device, wherein the sample placement device is used to place an original microtissue sample, and a hole with a preset aperture is opened on the sample placement device, the cutting device is used to cut the original microtissue sample set on the sample placement device, and the eddy current sampling device is arranged below the sample placement device, and is used to complete the collection of the cut microtissue sample by eddy current sampling of the microtissue sample released from the opening of the sample placement device after cutting.

[0008] Furthermore, the cutting device includes a high-precision laser device and a high-precision laser moving device, the high-precision laser device is arranged on the high-precision laser moving device, and the high-precision laser moving device is used to adjust the three-dimensional spatial position of the high-precision laser device.

[0009] Furthermore, the high-precision laser device can achieve focusing of the output light beam within a preset range.

[0010] Furthermore, the high-precision laser moving device includes a three-dimensional adjustment frame, which includes an X-axis adjustment frame, a Y-axis adjustment frame and a Z-axis adjustment frame, wherein the X-axis adjustment frame and the Y-axis adjustment frame serve to adjust the high-precision laser device over a large range above the cutting device, and the Z-axis adjustment frame is used to achieve a small adjustment in the height direction of the laser focus.

[0011] Furthermore, based on the adjustment of the high-precision laser moving device, the cutting of microtissue samples of specific shapes and / or the cutting of microtissue samples of arbitrary shapes are achieved.

[0012] Furthermore, the sample placement device is specifically a mesh-shaped loading platform, and the diameters of the holes opened on the loading platform are the same or different.

[0013] Furthermore, the vortex sampling device includes a vortex microtissue sample sampling tube, a negative pressure connecting tube and a liquid phase injection pump, the bottom end of the vortex microtissue sample sampling tube is connected to the negative pressure connecting tube, the negative pressure connecting tube is a three-way tube structure, wherein one output end of the negative pressure connecting tube is connected to the liquid phase injection pump, and the output end is connected to the outlet capillary.

[0014] Compared with the prior art, the present invention has the following advantages: the present invention realizes online extraction of samples cut by laser microscopy, realizes sampling under the flow process by means of liquid vortex and surface tension, and realizes sample transmission by means of liquid injection pump, which is convenient for subsequent analysis of microtissue samples. The present invention controls the microtissue sample to flow out to the outlet capillary by means of negative pressure, and the outlet capillary can process the microtissue sample by connecting other solvents. The process is complete, forming a one-stop sample pretreatment and sample analysis step, thereby improving experimental efficiency. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative labor.

[0016] Figure 1 It is a schematic diagram of the overall structure of the present invention.

[0017] Figure 2 This is a schematic diagram of cutting samples according to the present invention.

[0018] Figure 3Schematic diagram of exporting the cut sample through a three-way pipe.

[0019] Figure 4 It is the front view of the present invention.

[0020] Figure 5 Schematic diagram of the vortex tube structure of the present invention.

[0021] In the figure: 1. tissue sample loading platform; 2. three-dimensional adjustment frame; 3. laser head; 4. tissue section sample; 5. microtissue cutting sample; 6. vortex microtissue sample sampling tube; 7. connecting three-way pipe; 8. negative pressure connecting pipe; 9. liquid phase injection pump; 10. outlet capillary; 106. inner liner of vortex kit; 206. outer liner of vortex kit; 306. vortex internal output solution. DETAILED DESCRIPTION

[0022] It should be noted that, in the absence of conflict, the embodiments of the present invention and the features in the embodiments can be combined with each other. The present invention will be described in detail below with reference to the accompanying drawings and in combination with the embodiments.

[0023] In order to make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. The following description of at least one exemplary embodiment is actually only illustrative and is by no means intended to limit the present invention and its application or use. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0024] It should be noted that the terms used herein are only for describing specific embodiments and are not intended to limit exemplary embodiments according to the present invention. As used herein, unless the context clearly indicates otherwise, the singular form is also intended to include the plural form. In addition, it should be understood that when the terms "comprising" and / or "including" are used in this specification, it indicates the presence of features, steps, operations, devices, components and / or combinations thereof.

[0025] Unless otherwise specifically stated, the relative arrangement of the parts and steps described in these embodiments, the numerical expressions and numerical values ​​do not limit the scope of the present invention. At the same time, it should be clear that, for ease of description, the sizes of the various parts shown in the drawings are not drawn according to the actual proportional relationship. The technology, methods and equipment known to ordinary technicians in the relevant field may not be discussed in detail, but in appropriate cases, the technology, methods and equipment should be regarded as part of the authorization specification. In all examples shown and discussed here, any specific value should be interpreted as merely exemplary, rather than as a limitation. Therefore, other examples of exemplary embodiments may have different values. It should be noted that similar numbers and letters represent similar items in the following drawings, so once an item is defined in one drawing, it does not need to be further discussed in subsequent drawings.

[0026] In the description of the present invention, it is necessary to understand that the directions or positional relationships indicated by directional words such as "front, back, up, down, left, right", "lateral, vertical, perpendicular, horizontal" and "top, bottom" are usually based on the directions or positional relationships shown in the drawings. They are only for the convenience of describing the present invention and simplifying the description. Unless otherwise specified, these directional words do not indicate or imply that the device or element referred to must have a specific direction or be constructed and operated in a specific direction. Therefore, they cannot be understood as limiting the scope of protection of the present invention: the directional words "inside and outside" refer to the inside and outside relative to the contours of each component itself.

[0027] For ease of description, spatially relative terms such as "above", "above", "on the upper surface of", "above", etc. may be used here to describe the spatial positional relationship between a device or feature and other devices or features as shown in the figure. It should be understood that spatially relative terms are intended to include different orientations of the device in use or operation in addition to the orientation described in the figure. For example, if the device in the accompanying drawings is inverted, the device described as "above other devices or structures" or "above other devices or structures" will be positioned as "below other devices or structures" or "below their position devices or structures". Thus, the exemplary term "above" can include both "above" and "below". The device can also be positioned in other different ways (rotated 90 degrees or in other orientations), and the spatially relative descriptions used here are interpreted accordingly.

[0028] In addition, it should be noted that the use of terms such as "first" and "second" to limit components is only for the convenience of distinguishing the corresponding components. If not otherwise stated, the above terms have no special meaning and therefore cannot be understood as limiting the scope of protection of the present invention.

[0029] like Figures 1 to 5As shown, an embodiment of the present invention discloses an online eddy current sampling device for microtissue samples, comprising a cutting device, a sample placement device and an eddy current sampling device, wherein the sample placement device is used to place the original microtissue sample, and a hole with a preset aperture is opened on the sample placement device, the cutting device is used to cut the original microtissue sample 4 set on the sample placement device, and the eddy current sampling device is arranged below the sample placement device, and is used to complete the collection of the cut microtissue sample 5 released from the opening of the sample placement device through eddy current sampling.

[0030] Furthermore, the cutting device includes a high-precision laser device and a high-precision laser moving device, the high-precision laser device is arranged on the high-precision laser moving device, and the high-precision laser moving device is used to adjust the three-dimensional spatial position of the high-precision laser device.

[0031] Furthermore, the high-precision laser device can achieve focusing of the output light beam within a preset range. The core of the high-precision laser microdissection component is the laser light source, which has high energy and a small light spot. The laser head 3 can achieve focusing of a 50-5um light beam. During use, the laser light source remains stationary.

[0032] Furthermore, the high-precision laser moving device includes a three-dimensional adjustment frame 2, which includes an X-axis adjustment frame, a Y-axis adjustment frame and a Z-axis adjustment frame, wherein the X-axis adjustment frame and the Y-axis adjustment frame serve to adjust the high-precision laser device over a large range above the cutting device, and the Z-axis adjustment frame is used to achieve a small adjustment in the height direction of the laser focus.

[0033] Furthermore, based on the adjustment of the high-precision laser moving device, the cutting of microtissue samples of specific shapes and / or the cutting of microtissue samples of arbitrary shapes are achieved.

[0034] Furthermore, the sample placement device is specifically a mesh-shaped loading platform 1, and the apertures of the holes opened on the loading platform are the same or different.

[0035] Furthermore, the vortex sampling device includes a vortex microtissue sample sampling tube 6, a negative pressure connecting tube 8 and a liquid phase injection pump 9, the bottom end of the vortex microtissue sample sampling tube is connected to the negative pressure connecting tube, and the negative pressure connecting tube is a three-way tube 7 structure, wherein one output end of the negative pressure connecting tube is connected to the liquid phase injection pump, and the output end is connected to the outlet capillary 10.

[0036] Specifically, the microtissue samples after cutting are collected through a vortex microtissue sample sampling tube. In this embodiment, the vortex sampling tube is a sleeve structure, including an inner liner tube 106 of the vortex kit and an outer liner tube 206 of the vortex kit. The outer part is liquid outflow, and the inner part is liquid inflow, forming a vortex internal output solution 306, and the collected microtissue samples are collected into the liquid sampling tube. The inner diameter of the inner sampling tube is 150-200um, which is convenient for collecting microtissue samples with a diameter of about 100um after laser microcutting into the vortex sampling tube.

[0037] As an optional embodiment, the distance between the vortex sampling tube and the tissue sample loading platform can be adjusted. In order to facilitate the collection of microtissue samples during use, the vortex tube can be appropriately brought close to the laser microdissection instrument and appropriately adjusted to facilitate sample sampling.

[0038] When the laser microdissection component is working, the microtissue samples can be cut online by turning on the laser light source, and the microtissue samples with specific shapes can be cut by moving the three-dimensional adjustment frame, so that the tissue slice samples can be cut into any shape.

[0039] The vortex sampling tube absorbs the sample obtained from the top through surface tension, and the operation is divided into two processes. The first step is to sample the microtissue sample from the three-dimensional adjustment platform and enter the vortex microtissue sample sampling tube kit in the upper half. The second step is to inject the sample into the outlet capillary through the three-way connecting tube. The microtissue sample is sucked down from the tissue section, and the specific operation method is: using a liquid phase injection pump, the microtissue cutting sample is carried down from the top of the capillary by liquid flow through negative pressure, and enters the connecting three-way tube. At this time, the second step of the above operation is to control the flow of the microtissue sample to the outlet capillary by negative pressure, and the outlet capillary can be connected to other solvents to process the microtissue sample.

[0040] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. An online eddy current sampling device for microtissue samples, characterized in that: The invention comprises a cutting device, a sample placement device and an eddy current sampling device. The sample placement device is used to place the original microtissue sample, and a hole with a preset aperture is opened on the sample placement device. The cutting device is used to cut the original microtissue sample set on the sample placement device. The eddy current sampling device is arranged below the sample placement device and is used to complete the collection of the cut microtissue sample by eddy current sampling of the microtissue sample released from the opening of the sample placement device after cutting.

2. The micro-tissue sample online eddy current sampling device according to claim 1, characterized in that: The cutting device comprises a high-precision laser device and a high-precision laser moving device. The high-precision laser device is arranged on the high-precision laser moving device, and the high-precision laser moving device is used to adjust the three-dimensional spatial position of the high-precision laser device.

3. The micro-tissue sample online eddy current sampling device according to claim 2, characterized in that: The high-precision laser device can achieve focusing of the output light beam within a preset range.

4. The micro-tissue sample online eddy current sampling device according to claim 2, characterized in that: The high-precision laser moving device includes a three-dimensional adjustment frame, which includes an X-axis adjustment frame, a Y-axis adjustment frame and a Z-axis adjustment frame. The X-axis adjustment frame and the Y-axis adjustment frame are used to adjust the high-precision laser device over a large range above the cutting device, and the Z-axis adjustment frame is used to achieve a small adjustment in the height direction of the laser focus.

5. The micro-tissue sample online eddy current sampling device according to claim 1, characterized in that: Based on the adjustment of the high-precision laser moving device, the cutting of micro-tissue samples with specific shapes and / or the cutting of micro-tissue samples with arbitrary shapes are achieved.

6. The micro-tissue sample online eddy current sampling device according to claim 1, characterized in that: The sample placement device is specifically a mesh-shaped loading platform, and the apertures of the holes opened on the loading platform are the same or different.

7. The micro-tissue sample online eddy current sampling device according to claim 1, characterized in that: The vortex sampling device includes a vortex micro-tissue sample sampling tube, a negative pressure connecting tube and a liquid phase injection pump. The bottom end of the vortex micro-tissue sample sampling tube is connected to the negative pressure connecting tube. The negative pressure connecting tube is a three-way tube structure, wherein one output end of the negative pressure connecting tube is connected to the liquid phase injection pump, and the output end is connected to the outlet capillary.

Images