CN108593549A - A kind of sample handling device for microexamination - Google Patents

Abstract

The invention relates to a sample support device for microscopic observation, comprising a dish body and a sample support tube, two through holes are arranged on the side wall of the dish body, and the sample support tube passes through the two through holes Through the dish body and exposed head and tail outside the side wall of the dish body, the connection between the sample support tube and the through hole is a watertight connection, and the sample support tube can be wound around its central axis Turned, the sample support tube is open at both ends and provided with a removable stopper, and the end of the head is provided with a pipette connection. The sample support device of the present invention can be used to prepare samples conveniently and quickly, eliminating the complicated work of trimming the gel block after removing the sample preparation, and after sample preparation, the sample support tube can be rotated according to the needs of the experimenter, so that the sample can be conveniently placed The body posture is adjusted to the optimal state for observation. There will be no re-sampling due to incorrect sample posture.

Description

A sample support device for microscopic observation

technical field

The invention relates to the field of experimental equipment for microscopic observation, and more particularly relates to a sample support device for microscopic observation.

Background technique

The dynamic imaging of biological nerves in vivo is an important means of neurological research, and the imaging process requires the imaging system to have cellular-level resolution and a sufficiently high imaging rate. Since 2004, the light-sheet illumination microscope has developed very rapidly. It has achieved outstanding results in embryonic development research and neural research, and has gradually become an important means of scientific imaging. Light sheet illumination micro-imaging technology uses a light-speed thin sheet to excite fluorescence from the side of the sample, and obtains the fluorescence image at the illumination level through the microscope objective lens and CCD or sCOMS in the direction perpendicular to the light sheet, thus realizing the three-dimensional tomography of fluorescent samples imaging. Compared with confocal imaging technology, light sheet illumination microscopic imaging adopts surface detection imaging method instead of traditional point scanning imaging method, so it has higher imaging rate, higher signal-to-noise ratio and lower photobleaching. Toxic effects, suitable for imaging of living organisms for hours or even days.

At present, there are many manufacturers of light-sheet illumination microscopes on the market. Among them, Leica has developed the TwinFlect mirror and mounted the light-sheet system on the confocal platform. The unique TwinFlect mirror device deflects the illumination light sheet by 90 degrees, thereby Integrating illumination and detection optics into the longitudinal axis of each inverted Leica TCS SP8 without sacrificing confocal capabilities turns a confocal platform into a simple-to-use, versatile light-sheet microscope. Leica's light sheet systems also illuminate the sample in a single plane and are ideal for imaging sensitive samples and fast biological processes. Due to the absence of out-of-focus excitation, phototoxic effects can be localized to the focal plane. The sample to be observed is fixed and placed between the two lenses of the TwinFlect mirror, and the sample is moved under the light sheet to achieve 3D imaging of the sample.

Although the principles of light-sheet illumination microscopes from different manufacturers are the same, the sample preparation methods are completely different due to different hardware components. When observing small experimental animal samples such as zebrafish with the light sheet system of Leica Company, such experimental animal samples are generally fixed in a glass-bottomed Petri observation dish, and space is left on both sides of the sample so that the sample can be placed between two lenses.

At present, the sample preparation method for small experimental animal samples is as follows: first, a 1% agarose layer is laid on the bottom of the Petri dish as a cushion layer, and then 0.5% low-melting point agarose is added above the cushion layer and the sample is added to make the sample Maintain a specific posture and wait for the low-melting point agarose to solidify to form a sample immobilization layer. The agarose gel is then removed from both sides of the sample so that the fixed sample can be placed between the two lenses of the TwinFlect mirror. Finally, add water to the viewing vessel until the optics of the TwinFlect mirror can be submerged.

Due to the setting of the TwinFlect mirror, the light plane must be perfectly parallel to the sagittal plane of the experimental sample. The lens angle of the TwinFlect mirror is not adjustable, which makes the requirements for the body posture of the sample very strict (that is, the sagittal plane remains horizontal). As long as the sample is slightly offset when fixed, the obtained fixed sample will be Cannot be used for confocal scanning and must be re-prepared. However, the animal samples that can be observed by this instrument are all small samples such as zebrafish, and it is difficult to judge whether the sagittal plane is horizontal before going on the machine.

In addition, due to the need to remove the agarose gel on both sides of the sample during the preparation process, the contact area between the agarose gel and the bottom of the petri dish is relatively small. When adding water to the observation dish, the agarose gel that wraps the sample has May be washed away or float up.

The defect caused by the difficulty of sample preparation greatly limits the use of the device for observing animal samples, making the device underutilized.

Therefore, a new sample support device is needed to make sample preparation easier.

Contents of the invention

In order to solve the above problems, the present invention provides a sample support device for microscopic observation, including a dish body and a sample support tube, two through holes are arranged on the side wall of the dish body, and the sample support tube passes through The two through holes run through the dish body and expose the head and tail outside the side wall of the dish body. The connection between the sample support tube and the through hole is a watertight connection, and the sample support tube It can rotate around its own central axis, the two ends of the sample support tube are open, and are provided with detachable stoppers, and the end of the head is provided with a pipette connection.

The sample support device of the present invention can be used to prepare samples conveniently and quickly, eliminating the complicated work of trimming the gel block after removing the sample preparation, and after sample preparation, the sample support tube can be rotated according to the needs of the experimenter, and the sample can be conveniently placed Adjust the body posture to the optimal state for observation, and will not cause re-sampling due to incorrect sample body posture.

In a specific embodiment, a detachable through-hole plug made of elastic material is arranged in the through-hole of the dish body, the inner diameter of the through-hole plug matches the outer diameter of the sample support tube, and the The outer diameter of the through hole plug matches the diameter of the through hole of the dish. The elastic material can be rubber, elastomer, etc., and its specific form is not limited, as long as the sample support tube can pass through the through hole in a watertight manner and can rotate around the central axis.

Since samples such as zebrafish have different sizes, sample support tubes with different diameters are required for samples of different sizes. Through the above settings, the through-hole plug, sample support tube and dish body can be set in a detachable assembly form, and through-hole plugs with different inner diameters can be used to match sample support tubes with different diameters, and the assembly can be performed according to the size of the sample.

In a preferred embodiment, the head is also provided with a rotation adjustment part. Since the sample support tube is relatively thin, it is inconvenient to directly rotate the tube body of the sample support tube by hand during the rotation adjustment process. After the rotation adjustment part is provided, the rotation adjustment can be performed more conveniently and accurately.

Specifically, the rotation adjustment part is a handle provided on the head or the through-hole plug. For example, it is arranged on the head pipe body to form a handle with a larger diameter, and turning the handle makes the pipe body rotate relative to the through-hole plug. Or it is arranged on the through-hole plug, and the pipe body and the through-hole plug are rotated relative to the through-hole by rotating the handle. In order to realize the adjustment of the sample body posture.

In a preferred embodiment, an angle adjustment portion is further provided on the side wall of the vessel.

When the diameter of the tube body is too large relative to the sample, the body axis of the sample may form an included angle with the horizontal direction. At this time, the optimal body posture cannot be adjusted no matter how the sample support tube is rotated. If the angle adjustment part is used to adjust the angle between the tube body and the horizontal direction, the body axis of the sample is horizontal.

In a specific embodiment, the angle adjustment part is composed of a flexible connection part around the through hole, a cable and a shackle, one end of the cable is connected to the head of the sample support tube, and the other end is threaded through the passing through the shackle, and a positioning part is arranged on the pulling cable. The positioning part can be composed of multiple positioning blocks connected in series.

By pulling the cable, the angle between the pipe body and the horizontal direction can be adjusted conveniently, and the state can be fixed by the positioning knot. The angle adjustment part is not limited to the above structure, as long as the structure in the prior art can be used to adjust the angle between the sample support tube and the horizontal direction, it can be used as the angle adjustment part of the present invention.

In a preferred embodiment, the pipette connection is detachable. Different pipette connections are available depending on the pipette used.

In a preferred embodiment, the end opening of the tail is a smooth oblique cut. The smooth beveled cut increases the cross-sectional area of the tip, allowing for better aspiration of the sample without damaging it.

Description of drawings

Fig. 1 is the schematic diagram of an embodiment of the present invention;

Fig. 2 is a side view of an embodiment of the present invention;

Figure 3 is a side view of a sample support tube in one embodiment of the present invention;

Fig. 4 is a schematic structural view of the angle adjustment part, the dish body and the sample support tube in an embodiment provided with the angle adjustment part of the present invention.

In the accompanying drawings, the list of parts represented by each label is as follows:

1. Dish body, 2. Sample support tube, 11. Through-hole plug, 21. Head, 22. Tail, 23 Pipette connection part, 24. Rotation adjustment part, 121. Flexible connection part, 122. Cable, 123, shackle, 124, positioning part.

Detailed ways

The principles and features of the present invention are described below in conjunction with examples, which are only used to explain the present invention and are not intended to limit the scope of the present invention.

Example 1

As shown in Figure 1, the sample support device for microscopic observation in one embodiment includes a dish body 1 and a sample support tube 2, and two through holes are arranged on the side wall of the dish body 1, and the sample support The tube 2 runs through the dish body 1 through the two through holes and exposes a head 21 and a tail portion 22 outside the side wall of the dish body 1, and the connection between the sample support tube 2 and the through holes is a watertight connection , and the sample support tube 2 can be rotated around its own central axis, the two ends of the sample support tube 2 are open, and a detachable stopper is provided, and the end of the head 21 is provided with a pipette connection Section 23. The pipette connection part 23 is detachable. Different pipette connections are available depending on the pipette used. Pipettes usable in the present invention include pipettes, pipettes, pumps, etc., and are not limited to a specific form.

When in use, first immerse samples such as zebrafish in melted low-melting agarose, then connect the pipette connection part of the sample support device to the pipette, align the tail nozzle with the sample, and connect the sample to the agarose Pipet together into the sample support tube. The pipette is then removed and both ends of the sample support tube are stoppered. Place the sample support device horizontally and wait for the agarose to solidify. After the agarose gel is formed, the sample preparation is successful. It can be used for laser confocal scanning under the TwinFlect mirror of Leica Company. When on the machine, place the sample support device on the stage so that the sample support tube is located between the two lenses of the TwinFlect reflector, add water to the support device until the lens of the TwinFlect reflector can be submerged, and observe the sample Body posture, rotate the sample support tube to the coronal plane level of the sample, and the laser confocal scanning can be performed.

The sample support device of the present invention can be used to prepare samples conveniently and quickly, eliminating the complicated work of trimming the gel block after removing the sample preparation, and after sample preparation, the sample support tube can be rotated according to the needs of the experimenter, and the sample can be conveniently placed The body posture is adjusted to the optimal state for observation. There will be no re-sampling due to incorrect sample posture.

Example 2

As shown in Figure 2, in another embodiment, a detachable through-hole plug 11 made of elastic material is arranged in the through hole of the dish body 1, and the inner diameter of the through-hole plug 11 is consistent with the sample support. The outer diameter of the tube 2 is matched, and the outer diameter of the through hole plug 11 is matched with the diameter of the through hole of the dish body 1 . The elastic material can be rubber, elastomer, etc., and its specific form is not limited, as long as the sample support tube can pass through the through hole in a watertight manner and can rotate around the central axis.

Since samples such as zebrafish have different sizes, sample support tubes with different diameters are required for samples of different sizes. Through the above settings, the through-hole plug, sample support tube and dish body can be set in a detachable assembly form, and through-hole plugs with different inner diameters can be used to match sample support tubes with different diameters, and the assembly can be performed according to the size of the sample.

Example 3

As shown in FIG. 3 , in another preferred embodiment, the head 21 is further provided with a rotation adjustment part 24 , and the rotation adjustment part 24 is a handle provided on the head 21 . The end opening of the tail part 22 is an oblique cut. Since the sample support tube is relatively thin, it is inconvenient to directly rotate the tube body of the sample support tube by hand during the rotation adjustment process. After the rotation adjustment part is provided, the rotation adjustment can be performed more conveniently and accurately. It is arranged on the head pipe body to form a handle with a larger diameter, and turning the handle makes the pipe body rotate relative to the through-hole plug. The beveled cut increases the cross-sectional area of the tip, allowing for better sample aspiration.

In another embodiment, the rotation adjustment part 24 can be arranged on the through-hole plug, and the tube body together with the through-hole plug can be rotated relative to the through-hole by rotating the handle, so as to adjust the posture of the sample.

Example 4

As shown in FIG. 4 , in one embodiment, an angle adjustment portion is also provided on the side wall of the dish body 1 . When the diameter of the tube body is too large relative to the sample, the body axis of the sample may form an included angle with the horizontal direction. At this time, the optimal body posture cannot be adjusted no matter how the sample support tube is rotated. If the angle adjustment part is used to adjust the angle between the tube body and the horizontal direction, the body axis of the sample is horizontal.

Specifically, the angle adjustment part is composed of a flexible connection part 121 around the through hole, a cable 122 and a shackle 123. One end of the cable 122 is connected to the head 21 of the sample support tube, and the other end is Pass through the shackle 123 , and a positioning portion 124 is provided on the pull cable 122 . The positioning part can be composed of multiple positioning blocks connected in series.

By pulling the cable, the angle between the pipe body and the horizontal direction can be adjusted conveniently, and the state can be fixed by the positioning knot. The angle adjustment part is not limited to the above structure, as long as the structure in the prior art can be used to adjust the angle between the sample support tube and the horizontal direction, it can be used as the angle adjustment part of the present invention.

The above descriptions are only some embodiments of the present invention, and are not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention within.

Claims (8)

1. A sample support device for microscopic observation, characterized in that it comprises a dish body (1) and a sample support tube (2), the side wall of the dish body (1) is provided with two through holes, The sample support tube (2) runs through the dish body (1) through the two through holes and exposes the head (21) and tail (22) outside the side wall of the dish body (1). The connection between the support tube (2) and the through hole is a watertight connection, and the sample support tube (2) can rotate around its own central axis, the two ends of the sample support tube (2) are open, and the There is a removable stopper, the end of the head (21) is provided with a pipette connection (23). 2. The sample support device according to claim 1, characterized in that, a detachable through hole plug (11) made of elastic material is arranged in the through hole of the dish body (1), and the through hole plug The inner diameter of (11) matches the outer diameter of the sample support tube (2), and the outer diameter of the through hole plug (11) matches the diameter of the through hole of the dish body (1). 3. The sample supporting device according to claim 1, characterized in that a rotation adjustment part (24) is further provided on the head (21). 4. The sample supporting device according to claim 3, characterized in that, the rotation adjustment part (24) is a handle provided on the head (21) or the through-hole plug (11). 5. The sample supporting device according to claim 1, characterized in that, an angle adjustment part is further provided on the side wall of the dish body (1). 6. The sample support device according to claim 5, characterized in that, the angle adjustment part is composed of a flexible connection part (121) around the through hole, a cable (122) and a hook and loop (123), the One end of the cable (122) is connected to the head (21) of the sample support tube, and the other end passes through the shackle (123), and the cable (122) is provided with a plurality of positioning parts ( 124). 7. The sample supporting device according to any one of claims 1-6, characterized in that, the pipette connection part (23) is detachable. 8. The sample supporting device according to any one of claims 1-6, characterized in that, the end opening of the tail part (22) is an oblique cut.