CN215115888U - Object carrying glassware and fluorescent microscopic imaging device - Google Patents
Object carrying glassware and fluorescent microscopic imaging device Download PDFInfo
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- CN215115888U CN215115888U CN202120286226.8U CN202120286226U CN215115888U CN 215115888 U CN215115888 U CN 215115888U CN 202120286226 U CN202120286226 U CN 202120286226U CN 215115888 U CN215115888 U CN 215115888U
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Abstract
The utility model relates to a microscopic imaging technology field discloses a carry thing glassware and fluorescence microscopic imaging device, and wherein it includes to carry thing glassware: the body is provided with an object carrying surface for carrying a sample to be observed; and at least one fluorescent marker arranged on the object carrying surface, wherein the fluorescence excitation wavelength and the emission wavelength of the fluorescent marker are different from those of the fluorescent substance in the sample to be observed. By arranging the fluorescent marker, a reference standard is provided when the sample is observed, so that the region of interest observed before is conveniently and quickly searched. Meanwhile, the excitation wavelength and the emission wavelength of the fluorescent marker are set to be different from the characteristics of the fluorescent substance in the sample, so that contrast is formed, and the accuracy of the experiment is prevented from being influenced.
Description
Technical Field
The utility model relates to a microscopic imaging technology field specifically is a carry thing glassware and fluorescence microscopic imaging device.
Background
In a cell or tissue slice fluorescence imaging experiment, a sample needs to be placed on a fluorescence imaging device to observe an interested position, the imaging device is used for displaying a fluorescence image, and in addition, a camera device can be used for taking a picture so as to be convenient for scientific research papers to use. Some fluorescence imaging experiments may require viewing the same location before and after sample processing.
However, in the conventional fluorescence imaging experiment, a cell or tissue slice and other samples need to be placed on a transparent glass carrying vessel during observation, then the transparent glass carrying vessel with the samples is fixed on a carrying table, a proper magnification factor is selected, and the carrying table is continuously adjusted through a coarse adjustment mechanism and a fine adjustment mechanism so as to find the position to be observed in an enlarged view. The conventional glass carrier is a transparent glass slide or a transparent confocal glass, and the sample is cultured on the confocal glass or the tissue slice is fixed on the glass slide so as to be observed on the fluorescence imaging device. Before treatment, a sample is placed under a fluorescence microscopic imaging device, an interested position is found for observation, then after the sample is subjected to a corresponding treatment procedure outside the fluorescence microscopic imaging device, the sample is placed under the fluorescence microscopic imaging device again after the treatment is finished, the interested position is observed again, at the moment, because no mark for reference exists, the observation is time-consuming and labor-consuming, and the searching success rate is low. Existing traditional practice will adopt to carry out invasive mark to the sample, for example will carry the cell population that the sample needs the observation position in the glassware to destroy, make it form acellular blank mark to seek under the microscopic imaging device of fluorescence, but this kind of mark mode causes whole sample to pollute easily, destroys the sample state, and then influences the accuracy of experimental result, because experimental treatment needs to incubate for a long time, blank mark position may be full of cells once more, leads to seeking difficultly.
SUMMERY OF THE UTILITY MODEL
The utility model aims to solve the technical problem that overcome traditional fluorescence microscopic imaging device among the prior art in cell fluorescence imaging experiment, when the same position around needs observation processing, owing to lack the marker, the operation is wasted time and energy and the lower technical problem of success rate to a fluorescence microscopic imaging device who carries thing glassware and have this year thing glassware with marker is provided.
To this end, one of the objects of the present invention is to provide a glass container, comprising:
the body is provided with an object carrying surface for carrying a sample to be observed;
and the fluorescent label is arranged on the carrying surface, and the fluorescence excitation wavelength and the emission wavelength of the fluorescent label are different from those of the fluorescent substance in the sample to be observed.
Optionally, the glass carrier includes at least two crossed fluorescent mark portions, and the fluorescent materials of the two fluorescent mark portions have different emission wavelengths under the same excitation light.
Optionally, the fluorescent marker is cross-shaped or m-shaped.
Optionally, the loading glass vessel is adapted to fix the fluorescent marker at the geometric center of the loading surface.
Optionally, the glass carrier comprises a plurality of fluorescent markers regularly arranged.
Optionally, in the glass container, the fluorescent marker is a thin film fluorescent sticker with one surface provided with fluorescence, and the surface provided with the fluorescence is stuck on the object carrying surface; or
And a positioning surface on the opposite side of the object carrying surface is provided with a slot which is recessed towards the object carrying surface from the positioning surface, and the fluorescent marker is inserted and fixed in the slot.
Another object of the present invention is to provide a fluorescence microscopic imaging apparatus, including:
an excitation light module for generating a laser beam as excitation light;
the electronic control object stage comprises an operation table, an object stage which can be controlled by the operation table to move on a horizontal plane and a rotating module which is arranged on the object stage and can be controlled by the operation table to rotate relative to the object stage, wherein the operation table can display the plane displacement of the object stage and the rotating angle of the rotating module;
the object carrying glass vessel is any one of the object carrying glass vessels, is fixed on the rotating module and follows the rotating module, and is used for placing a sample to be observed, and the laser beam is focused on the fluorescent marker on the object carrying glass vessel and the sample to be observed;
the image acquisition module is suitable for acquiring fluorescence images generated by respectively exciting the fluorescence marker and the sample to be observed by the laser beam;
and the computer is connected with the image acquisition module and is used for displaying the fluorescence image.
Optionally, the fluorescence microscopy imaging apparatus, the rotation module includes:
the rotating piece is arranged on the objective table, and a sheet pusher structure used for fixing and moving the objective glassware is arranged on the rotating piece;
and the rotating control part is electrically connected with the operating platform and the rotating part and is suitable for being controlled by the operating platform to drive the rotating part to rotate.
Optionally, in the fluorescence microscopic imaging apparatus, the plane displacement of the object stage and the rotation angle of the rotation module may be displayed on a display screen of the console or on a software interface of a computer.
The utility model discloses technical scheme has following advantage:
1. the utility model discloses a carry thing glassware, include: the body is provided with an object carrying surface for carrying a sample to be observed;
and the fluorescent label is arranged on the carrying surface, and the fluorescence excitation wavelength and the emission wavelength of the fluorescent label are different from those of the fluorescent substance in the sample to be observed.
By arranging the fluorescent marker, a reference standard is provided when the sample is observed, so that the region of interest observed before is conveniently and quickly searched. Meanwhile, the excitation wavelength and the emission wavelength of the fluorescent marker are set to be different from the characteristics of the fluorescent substance in the sample, so that contrast is formed, and the accuracy of the experiment is prevented from being influenced.
2. The utility model discloses a carry thing glassware, the fluorescence marker includes two at least and is the crossing fluorescence mark portion of cross and two the fluorescent substance of fluorescence mark portion emission wavelength under same exciting light is different. So that the directions of the X direction and the Y direction of the horizontal plane coordinate can be accurately determined, and the problem that the directions cannot be determined when the horizontal plane coordinate is observed again is avoided.
3. The fluorescence microscopic imaging device of the utility model comprises an excitation light module, a fluorescence microscopic imaging module and a fluorescence microscopic imaging module, wherein the excitation light module is used for generating a laser beam as excitation light;
the electronic control object stage comprises an operation table, an object stage which can be controlled by the operation table to move on a horizontal plane and a rotating module which is arranged on the object stage and can be controlled by the operation table to rotate relative to the object stage, wherein the operation table can display the plane displacement of the object stage and the rotating angle of the rotating module;
the object carrying glass vessel is any one of the object carrying glass vessels, is fixed on the rotating module and follows the rotating module, and is used for placing a sample to be observed, and the laser beam is focused on the fluorescent marker on the object carrying glass vessel and the sample to be observed;
the image acquisition module is suitable for acquiring fluorescence images generated by respectively exciting the fluorescence marker and the sample to be observed by the laser beam;
and the computer is connected with the image acquisition module and is used for displaying the fluorescence image. Due to the glass vessel with the fluorescent marker, the origin can be calibrated by the fluorescent marker during cell experiment observation. But through setting the objective table to horizontal migration, collocation rotation module for place the sample of waiting to observe in the year thing glassware on the objective table can remove and rotate at the horizontal plane, operate and show horizontal displacement volume and rotation angle through the operation panel, so that when observing once more, the horizontal displacement volume and the rotation angle that can show carry out quick pursuit location to observing the position, labour saving and time saving, convenient operation, the success rate is higher.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the embodiments or the technical solutions in the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to these drawings without creative efforts.
Fig. 1 is a schematic structural diagram of one of the glasses for carrying objects according to embodiment 1 of the present invention;
fig. 2 is a schematic structural view of another loading glass vessel according to embodiment 1 of the present invention;
FIG. 3 is an enlarged schematic view of the fluorescent marker of the loading glass of FIGS. 1 and 2;
FIG. 4 is a front cross-sectional structural schematic view of the carrier glassware of FIG. 1;
FIG. 5 is a front cross-sectional structural schematic view of the carrier glassware of FIG. 2;
FIG. 6 is a schematic cross-sectional enlarged view of the fluorescent marker of the loading glass of FIGS. 4 and 5;
fig. 7 is a schematic top view of an electronic control stage of a fluorescence microscopic imaging apparatus according to embodiment 2 of the present invention;
fig. 8 is a schematic structural diagram of the operation console in embodiments 2 and 3 of the present invention (including the adjustment button for the horizontal displacement and the rotation angle, and the display screen for the horizontal displacement and the rotation angle).
Description of reference numerals:
1. a body; 10. a carrying surface; 11. a fluorescent label; 110. a first fluorescent labeling unit; 111. a second fluorescent labeling unit; 12. a groove;
2. an object stage;
3. a rotation module;
4. a blade pusher;
5. an operation table;
6. an X-direction adjusting knob; 61. an X-direction displacement display screen;
7. a Y-direction adjusting knob; 71. a Y-direction displacement display screen;
8. a rotation angle adjusting knob; 81. the display screen is rotated.
Detailed Description
The technical solution of the present invention will be described clearly and completely with reference to the accompanying drawings, and obviously, the described embodiments are some, but not all embodiments of the present invention. Based on the embodiments in the present invention, all other embodiments obtained by a person skilled in the art without creative work belong to the protection scope of the present invention.
Example 1
Referring to fig. 1 to 6, the present embodiment provides a carrier glass, including: the fluorescent microscope comprises a body 1 and at least one fluorescent marker 11, wherein the body 1 is provided with an opposite object carrying surface 10 and a positioning surface (not marked), the object carrying surface 10 is suitable for placing a sample (not shown) to be observed and arranging the fluorescent marker 11, and the positioning surface is suitable for fixing and positioning the object carrying glassware on an electric control object stage of a fluorescent microscope imaging device. The body 1 of the carrier glass vessel of the present embodiment is a conventional confocal vessel (as shown in fig. 1 and 4) or a glass slide (as shown in fig. 2 and 5). The excitation wavelength and the emission wavelength of the fluorescent marker 11 are different from those of the fluorescent substance of the sample to be observed, and are not specifically described or limited, and those skilled in the art can select the fluorescent marker 11 with different excitation wavelengths and emission wavelengths according to the characteristics of the fluorescent substance of the sample to be observed, so that the fluorescent marker 11 can not interfere with the sample while being clearly positioned by the fluorescent marker, and the accuracy of the experiment is high.
As shown in fig. 3 and 5, the fluorescent marker 11 includes at least two crossed fluorescent markers, and for convenience of description and distinction, the transverse fluorescent marker shown in fig. 3 is described as the first fluorescent marker 110, and the vertical fluorescent marker shown in fig. 3 is described as the second fluorescent marker 111. It is preferable that the fluorescent substances of the first fluorescent labeling unit 110 and the second fluorescent labeling unit 111 have different emission wavelengths under the same excitation light. The fluorescent substances arranged in a cross shape and having different emission wavelengths under the same excitation light of the first fluorescent labeling portion 110 and the second fluorescent labeling portion 111 are aimed at determining the lateral direction, i.e., the X direction, and the longitudinal direction, i.e., the Y direction, of the orthogonal coordinate system of the horizontal plane, avoiding the problem that the direction cannot be determined when using, for example, a circular confocal dish or a square slide glass. The fluorescent marker 11 may have a cross shape as shown in fig. 3, or may have other shapes such as a cross shape like a Chinese character mi or an L shape. The number of fluorescent markers 11 may be a cross or a meter, or a cross or a meter or an L, which is formed by a plurality of marker portions and has a gap between the marker portions.
The shape of the fluorescent marker 11 in the present embodiment is preferably a normal cross shape. As shown in fig. 3, the cross-shaped fluorescent marker is composed of a first fluorescent marker portion 110 along the transverse direction, i.e., the X-axis direction, and a second fluorescent marker portion 111 along the longitudinal direction, i.e., the Y-axis direction, the lengths of the first fluorescent marker portion 110 and the second fluorescent marker portion 111 are both preferably 100 μm, and the widths of the first fluorescent marker portion 110 and the second fluorescent marker portion 111 are both preferably 10 μm, and it should be noted that the lengths and the widths of the first fluorescent marker portion 110 and the second fluorescent marker portion 111 are not limited in detail, and those skilled in the art can reasonably select and design according to the imaging magnification.
The fluorescent marker 11 may be disposed at a geometric center of the carrier surface 10 of the glass carrier, such as a center of a confocal dish or an intersection of diagonals of a glass slide, but may be disposed at any other position of the carrier surface instead of the geometric center. Preferably at the geometric center.
The number of the fluorescent markers 11 is preferably one, and may be other numbers, such as two, four, eight, etc., and two are formed in an L-shape with a hollow intersection. Four crosses formed as a positive intersection with a central void, i.e., four fluorescent markers do not intersect. Eight forms a rice-shaped body with a hollow center. As long as it is ensured that the planar coordinate directions, i.e., the X direction and the Y direction, can be determined and that matching with the standard cross mark portion on the screen can be performed with coincidence is achieved, so that accurate positioning of the origin can be achieved.
For the fixing mode of the fluorescent marker 11 and the object-carrying glass vessel, the selectable fluorescent marker 11 is a thin film fluorescent sticker with one surface provided with fluorescence, good cohesiveness and sealing performance, and the surface provided with fluorescence is pasted on the object-carrying surface 10. As shown in fig. 4, a circular confocal dish is formed, a groove 12 is formed in the middle of the object carrying surface, i.e. the inner bottom surface of the confocal dish, a slot is formed in the outer side of the bottom surface of the groove, i.e. the positioning surface opposite to the object carrying surface, and the slot is recessed from the positioning surface toward the object carrying surface, i.e. from the outer side of the bottom surface toward the inner side of the bottom surface, and the fluorescent marker is inserted into the slot. Optionally, the width of the slot is 1/4 of the diameter of the object carrying surface 10, specifically the bottom wall of the confocal dish, for example, the diameter of the confocal dish is 35mm, the width of the groove 12 is 8.5mm, and the thickness is 1/2 mm of the thickness of the glass ware, that is, 0.9 mm.
Example 2
The fluorescence microscopic imaging apparatus provided in this embodiment, referring to fig. 1 to 8, includes an excitation light module (not shown), an electrically controlled stage, the objective glassware of embodiment 1, an image acquisition module, and a computer. Wherein the excitation light module is used for generating laser beams as excitation light; the electric control object stage comprises an operation table 5, an object stage 2 which can be controlled by the operation table 5 to move on a horizontal plane, and a rotating module 3 which is arranged on the object stage 2 and can be controlled by the operation table 5 to rotate relative to the object stage 2, wherein the operation table 5 can display the plane displacement of the object stage 2 and the rotating angle of the rotating module 3; the object carrying glass ware is fixed on the rotating module 3 and follows the rotating module 3, and is used for placing a sample to be observed, and the laser beam is focused on the fluorescent marker 11 on the object carrying glass ware and the sample to be observed; the image acquisition module is suitable for acquiring fluorescence images generated by exciting the fluorescence marker 11 and the sample to be observed by the laser beam respectively; and the computer is connected with the image acquisition module and is used for displaying the fluorescence image.
For the electrically controlled object stage, as shown in fig. 7, the object stage 2 is a conventional object stage capable of moving horizontally, specifically, moving horizontally and longitudinally, on a conventional fluorescence microscopic imaging device, the rotating module 3 is a cylindrical rotating disk, a conventional push piece device 4 for fixing and moving an object glass ware is arranged in the middle, the structure of the push piece device 4 is consistent with that of the push piece device on the conventional fluorescence microscopic imaging device, and the operation principle is the same, and it should be noted that the push piece device 4 can be detached and replaced. The operation panel 5 is an existing conventional PLC operation panel, and can operate and control the movement of the object stage 2 on the horizontal plane and the rotation of the rotating module 3 relative to the object stage 2 on the horizontal plane, as shown in fig. 8, it can be an X-direction adjusting knob 6, a Y-direction adjusting knob 7 or a rotation angle adjusting knob 8, which are used for adjusting the X-direction displacement of the object stage 2, the Y-direction displacement and adjusting the rotation angle of the rotating module 3, respectively, and can be a virtual adjusting knob or a physical adjusting knob directly arranged on the operation panel 5, and the operation panel 5 can also be provided with an X-direction displacement S for realizing the movement of the object stage 2XAnd amount of Y-directional displacement SYAnd the rotation angle theta of the rotation module 3, and the operation table can also realize the displacement SX、SYAnd the zero setting operation of the rotation angle theta is realized in a specific mode and principle of zero setting of a timer in the existing mobile phone softwareIn a consistent manner and not described or limited in detail.
For the rotating module 3, the rotating module comprises a rotating piece and a rotating control piece, wherein the rotating piece is arranged on the object stage 2 and is used for installing the blade pushing device 4; the slide glass vessel containing the sample to be observed is fixed, moved and positioned by the slide pusher 4. The rotation control member is electrically connected with the operation table 5 and the rotating member and is suitable for controlling and driving the rotating member to rotate through the operation table 5. The rotation control member may be a conventional rotating electric machine, and the rotation member is a rotating disk or a rotating column connected to an output shaft of the rotating electric machine, and will not be described and limited in detail.
The display of the plane displacement of the object stage 2 and the rotation angle of the rotation module 3 can be directly displayed on the display interface of the console 5, such as the X-direction displacement display screen 61, the Y-direction displacement display screen 71 and the rotation angle display screen 81 shown in fig. 8, or can be directly displayed on the imaging software interface of the computer. It should be noted that, if the standard cross displayed on the imaging software interface is adopted, a concealable standard cross can be added on the imaging software interface for matching with the cross of the fluorescent marker on the glass vessel for carrying the object so as to calibrate the origin, and the standard cross marker can be concealed after calibration is completed.
The use method provided by the embodiment comprises the following steps:
s1, fixing a sample to be observed on a rotating module 3 of an electric control objective table;
s2, finding a fluorescent marker on the glass vessel in the visual field with corresponding magnification, judging whether the fluorescent marker in the visual field is matched with a standard cross of a display screen or an imaging software interface, if not, adjusting the horizontal displacement and the rotation angle of the electric control objective table through an operation table to enable the fluorescent marker to be coincided and matched with the standard cross, and then zeroing the displacement and the rotation angle of the electric control objective table displayed at the moment to calibrate the position of an original point;
s3, adjusting the electric control objective table, finding a target observation area of the sample to be observed in the visual field, and recording the displacement and the rotation angle displayed at the moment;
and S4, taking down the sample to be observed, carrying out corresponding treatment, putting the sample on the object glass ware on the electric control object stage again, repeating the step S2, adjusting the electric control object stage according to the displacement and the rotation angle recorded in the step S3, finding the region observed before treatment again, and observing again.
By adopting the using method, when the same position of the sample to be observed needs to be observed again after the treatment, the displacement and the rotation angle displayed on the display screen or the interface of the imaging software can be directly and quickly positioned and tracked according to the observation position recorded before the treatment, so that the searching time can be greatly saved, the time and the labor are saved, and the success rate is higher.
The above, only be the concrete implementation of the preferred embodiment of the present invention, but the protection scope of the present invention is not limited thereto, and any person skilled in the art is in the technical scope of the present invention, according to the technical solution of the present invention and the utility model, the concept of which is equivalent to replace or change, should be covered within the protection scope of the present invention.
Claims (9)
1. A carrier glassware, comprising:
a body (1) having a loading surface (10) for loading a sample to be observed;
at least one fluorescent marker (11) is arranged on the object carrying surface (10), and the fluorescent excitation wavelength and the emission wavelength of the fluorescent marker (11) are different from those of a fluorescent substance in a sample to be observed.
2. A carrier glass according to claim 1, wherein the fluorescent marker (11) comprises at least two crossed fluorescent markers and the fluorescent materials of the two fluorescent markers emit different wavelengths under the same excitation light.
3. A carrier glass according to claim 1 or 2, characterised in that the fluorescent marker (11) is cross-shaped or m-shaped.
4. An object carrying glass according to claim 1 or 2, wherein the fluorescent marker (11) is adapted to be fixed in a geometrically central position of the object carrying surface (10).
5. A carrier glass according to claim 1, wherein said fluorescent marker (11) comprises a plurality of fluorescent markers (11), and said plurality of fluorescent markers (11) are arranged regularly.
6. A carrier glass according to claim 1 or 2, wherein the fluorescent marker (11) is a thin film fluorescent sticker with fluorescence on one side, and the side with the fluorescence is adhered to the carrier surface (10); or
And a positioning surface at the opposite side of the object carrying surface (10) is provided with a slot which is concave towards the object carrying surface (10) from the positioning surface, and the fluorescent marker (11) is inserted and fixed in the slot.
7. A fluorescence microscopy imaging device, comprising:
an excitation light module for generating a laser beam as excitation light;
the electronic control object stage (2) comprises an operation table (5), an object stage (2) which can be controlled by the operation table (5) to move on a horizontal plane and a rotating module (3) which is arranged on the object stage (2) and can be controlled by the operation table (5) to rotate relative to the object stage (2), wherein the operation table (5) can display the plane displacement of the object stage (2) and the rotating angle of the rotating module (3);
-a carrier glass according to any of claims 1 to 6, fixed to the rotary module (3) and following the rotary module (3) for placing a sample to be observed, the laser beam being focused on the fluorescent marker (11) on the carrier glass and on the sample to be observed;
the image acquisition module is suitable for acquiring fluorescence images generated by respectively exciting the fluorescence marker (11) and the sample to be observed by the laser beam;
and the computer is connected with the image acquisition module and is used for displaying the fluorescence image.
8. The fluorescence microscopy imaging device according to claim 7, characterized in that the rotation module (3) comprises:
the rotating piece is arranged on the object stage (2), and a blade pusher (4) structure for fixing and moving the object-carrying glassware is arranged on the rotating piece;
and the rotating control part is electrically connected with the operating platform (5) and the rotating part and is suitable for being controlled by the operating platform (5) to drive the rotating part to rotate.
9. The fluorescence microscopy imaging device according to claim 7 or 8, characterized in that the planar displacement of the object stage (2) and the rotation angle of the rotation module (3) can be displayed on a display screen of the console (5) or on a software interface of a computer.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202120286226.8U CN215115888U (en) | 2021-01-29 | 2021-01-29 | Object carrying glassware and fluorescent microscopic imaging device |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202120286226.8U CN215115888U (en) | 2021-01-29 | 2021-01-29 | Object carrying glassware and fluorescent microscopic imaging device |
Publications (1)
| Publication Number | Publication Date |
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| CN215115888U true CN215115888U (en) | 2021-12-10 |
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| CN202120286226.8U Active CN215115888U (en) | 2021-01-29 | 2021-01-29 | Object carrying glassware and fluorescent microscopic imaging device |
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