CN117270186A - Multi-angle shooting assembly, shooting system and shooting method for microscope sample - Google Patents
Multi-angle shooting assembly, shooting system and shooting method for microscope sample Download PDFInfo
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- CN117270186A CN117270186A CN202311276350.6A CN202311276350A CN117270186A CN 117270186 A CN117270186 A CN 117270186A CN 202311276350 A CN202311276350 A CN 202311276350A CN 117270186 A CN117270186 A CN 117270186A
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Classifications
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- G—PHYSICS
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- G02B21/00—Microscopes
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- G02B21/00—Microscopes
- G02B21/36—Microscopes arranged for photographic purposes or projection purposes or digital imaging or video purposes including associated control and data processing arrangements
Abstract
The invention relates to the technical field of microscopes, in particular to a component, a shooting system and a shooting method for shooting a sample at multiple angles for a microscope.
Description
Technical Field
The invention relates to the technical field of microscopes, in particular to a component for shooting a sample at multiple angles for a microscope, a shooting system and a shooting method.
Background
Most objects have a three-dimensional structure, and thus it is necessary to observe the objects from multiple angles in order to accurately describe their three-dimensional structure. Taking three-dimensional cytospheres as an example, most laboratories observe the morphology of three-dimensional cytospheres extremely dependent on inverted microscopes. Since three-dimensional cell spheres and two-dimensional growing cells produce a three-dimensional morphological change in one more dimension, only the projection of the three-dimensional cell spheres in the Z direction (perpendicular to the tabletop direction) can be observed using a conventional inverted microscope. If the side morphology of the three-dimensional cell sphere needs to be continuously observed, a confocal microscope is usually selected to shoot the fluorescence distribution of the three-dimensional cell sphere, and the monolayer profile of the cell sphere is observed by embedding the section to estimate the three-dimensional morphology or the surface of the cell sphere is observed by a scanning electron microscope. However, the conventional photographing means generally need to perform steps such as embedding, metal spraying and sample preparation, so that not only the true state of the cell ball cannot be maintained, but also the cell ball is dead after photographing, cannot continue to grow, and cannot observe in real time and continuously, so that the exploration of the dynamic process of the growth of the cell ball is fatal. At the same time, the photographic effect is not enough to fully exhibit the complete side morphology of the photographed sample: the bright field morphology image and the cell distribution image on the cell pellet cannot be captured at the same time.
Meanwhile, for samples such as two liquid phase separation interfaces, which can not directly turn over the samples to perform liquid level layering observation, the two-phase intersection interface can not be directly observed through optical observation equipment such as a microscope. Similarly, the real-time observation and monitoring of the film coating interface and the observation and monitoring of the hydrogen production efficiency of the surface of the electrocatalytic material are difficult to directly carry out in the process of sample film coating.
Disclosure of Invention
The invention aims to provide a component, a shooting system and a shooting method for shooting a sample for a microscope at multiple angles, and aims to solve the problem that the conventional microscope cannot realize the observation of the sample on the microscope at multiple angles.
In order to achieve the above object, in a first aspect, the present invention provides an assembly for multi-angle photographing of a sample for a microscope, comprising a supporting device body and a reflecting mirror, wherein the reflecting mirror is connected with one side of the supporting device body. And reflecting images of different surfaces of the sample by the reflecting mirror, observing the images in the reflecting mirror by using a microscope, and photographing, so that the sample is photographed at multiple angles on the premise of not changing the postures of the microscope objective lens and the sample.
In some embodiments, the support device body includes a housing and a lighting module, the housing has a sample placement slot, and the lighting module and the reflector are both connected with an inner sidewall of the sample placement slot. In order to make the observation effect better, use the lighting module to gather light to use the influence of ambient light is reduced to the casing.
The microscope can be classified into an upright microscope and an inverted microscope according to the positional relationship between the objective lens and the observed sample. Wherein the objective lens of the front microscope is positioned above the observed sample, and the observation is carried out from the upper part; the objective lens of the inverted microscope is positioned below the observed sample, and the observation is performed from below.
For a positive microscope, in some embodiments, the housing has an objective lens capture port in communication with the sample well and above the mirror. The upright microscope is more convenient to observe through the objective shooting port.
For an inverted microscope, in some embodiments, the support device body further comprises a glass sheet detachably connected to the housing and disposed below the reflector and the illumination module. The glass sheet is used for placing a sample.
Wherein the housing has a dimmer switch disposed on the housing.
The lighting assembly comprises a light source fixing base, a spotlight rotating hinge, a spotlight shell, a lamp bead and a lens, wherein the spotlight fixing base is fixedly connected with the inner side wall of the sample placing groove, the spotlight rotating hinge is fixedly connected with the spotlight fixing base and is located on one side of the spotlight fixing base, the spotlight shell is fixedly connected with the spotlight rotating hinge and is located on one side of the spotlight fixing base, the spotlight bead is fixedly connected with the spotlight shell and is located in the spotlight shell, and the lens is fixedly connected with the spotlight shell and is located on one side of the spotlight shell.
In some embodiments, the supporting device body further comprises a base, a reflector supporting frame, a rotating fixing support and a reflector fixing table, wherein the base is fixedly connected with one end of the reflector supporting frame, the other end of the reflector supporting frame is provided with a rotating shaft, one end of the rotating fixing support is movably connected with the rotating shaft, the reflector fixing table is movably connected with the other end of the rotating shaft fixing frame, the reflector is fixedly connected with the side face of the reflector fixing table, and the reflector fixing table is further provided with a rotating angle observation hole. The supporting device is convenient to operate by rotating and adjusting the angle of the reflecting mirror.
In a second aspect, a specimen multi-angle photographing system for a microscope includes the specimen multi-angle photographing assembly for a microscope of the first aspect,
the microscope comprises a plane of the reflecting mirror, and is characterized by further comprising a microscope objective, wherein the included angle between the optical central axis of the microscope objective and the plane of the reflecting mirror is between 0 and 90 degrees, and is not equal to 0 or 90 degrees.
In a third aspect, a method for multi-dimensionally photographing a sample for a microscope, applied to the assembly for multi-dimensionally photographing a sample for a microscope according to the first aspect, includes the steps of:
sequentially reflecting images of different surfaces of a sample by the reflector 1;
the image of 1 in the mirror is captured sequentially by a microscope and photographed.
The invention relates to a multi-dimensional shooting assembly, a shooting system and a shooting method for a sample for a microscope. The supporting device is used for fixing the reflecting mirror, then the reflecting mirror is placed on the objective table, when the side surface of a sample is needed, the reflecting mirror is placed above the microscope objective, the sample is placed on the side surface of the reflecting mirror, the angle and the sample position of the reflecting mirror are adjusted, so that the reflecting mirror can display the image of the sample, then the relative position between the reflecting mirror and the microscope objective is adjusted through a moving device of a microscope platform, the image of the side surface of the sample in the reflecting mirror is captured by the objective, the side surface of the sample can be observed, the observation of the same sample at different angles is realized, and therefore, the bottom surface and the side surface of the sample are photographed by using photographing equipment. The invention does not need special surface treatment so as to realize effective cost control and reduce the difficulty of sample preparation technology of multi-angle observation; the sample is not required to be contacted, so that the sample pollution is reduced to the greatest extent; meanwhile, expensive facility equipment is not needed, the threshold for multi-angle observation is reduced, observation and shooting can be carried out through a laboratory conventional microscope, and the problem that the conventional microscope cannot realize multi-angle morphological observation of a sample on the microscope is solved.
Drawings
In order to more clearly illustrate the embodiments of the invention or the technical solutions in the prior art, the drawings that are required in the embodiments or the description of the prior art will be briefly described, it being obvious that the drawings in the following description are only some embodiments of the invention, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
Fig. 1 is a schematic structural view of a first embodiment of a multi-angle photographing assembly for a microscope sample according to the present invention.
Fig. 2 is a schematic view of another direction of the first embodiment of the multi-angle photographing assembly for a microscope provided by the present invention.
Fig. 3 is a cross-sectional view of a first embodiment of a sample multi-angle photographic assembly for a microscope provided by the present invention.
Fig. 4 is a schematic view of the optical path of the first embodiment of the present invention.
Fig. 5 is a schematic structural view of a second embodiment of a multi-angle photographing assembly for a microscope sample according to the present invention.
Fig. 6 is a cross-sectional view of a second embodiment of a sample multi-angle photographic assembly for a microscope provided by the present invention.
Fig. 7 is a schematic structural view of a third embodiment of a multi-angle photographing assembly for a microscope sample according to the present invention.
Fig. 8 is a bright field image of the side and bottom surfaces of the three-dimensional cell sphere of example 1 in the first embodiment.
Fig. 9 is a fluorescent image of the bright field and dio+dapi staining of the side and bottom surfaces of the three-dimensional cell sphere of example 2 of the first embodiment.
Fig. 10 is a bright field image of the liquid contact angle of the surface of the material of example 3 of the first embodiment.
Fig. 11 is an oil liquid contact angle bright field image of the underwater oleophobic of the material of example 4 of the first embodiment.
Fig. 12 is a bright field image of hydrogen bubbles generated on the electrode surface in the aqueous electrolytic hydrogen production of example 5 of the first embodiment.
Fig. 13 is a thermal fused deposition printing of a different angle side microstructure of an example in a third embodiment.
Fig. 14 is a schematic diagram of a sample multi-angle shooting system for a microscope provided by the invention.
Fig. 15 is a flowchart of a method for photographing a sample for a microscope at multiple angles according to the present invention.
FIG. 16 is a schematic view of the structure of a liquid sample cell.
1-reflector, 2-shell, 3-sample placing groove, 4-objective shooting port, 5-shot lamp fixed base, 6-shot lamp rotary hinge, 7-shot lamp shell, 8-lamp pearl, 9-lens, 10-base, 11-reflector support frame, 12-rotation fixed support, 13-reflector fixed platform, 14-rotation angle observation hole, 15-glass piece, 16-dimmer switch, 17-liquid sample groove, 18-transparent shell, 19-optical glass piece.
Detailed Description
Embodiments of the present invention are described in detail below, examples of which are illustrated in the accompanying drawings, wherein like or similar reference numerals refer to like or similar elements or elements having like or similar functions throughout. The embodiments described below by referring to the drawings are illustrative and intended to explain the present invention and should not be construed as limiting the invention.
Referring to fig. 1 to 15, in a first aspect, the present invention provides an assembly for multi-angle photographing of a sample for a microscope,
first embodiment:
the device comprises a supporting device body and a reflecting mirror 1, wherein the reflecting mirror 1 is arranged on one side of the supporting device body.
The support device body comprises a shell 2 and an illumination assembly, the shell 2 is provided with a sample placing groove 3, the illumination module is arranged on the inner side wall of the sample placing groove 3, and the reflecting mirror 1 is also arranged on the inner side wall of the sample placing groove 3.
The lighting module comprises a spotlight fixing base 5, a spotlight rotating hinge 6, a spotlight shell 7, a lamp bead 8 and a lens 9, wherein the spotlight fixing base 5 is fixedly connected with the inner side wall of the sample placing groove 3, the spotlight rotating hinge 6 is fixedly connected with the spotlight fixing base 5 and is positioned on one side of the spotlight fixing base 5, the spotlight shell 7 is fixedly connected with the spotlight rotating hinge 6 and is positioned on one side, far away from the spotlight fixing base 5, of the spotlight rotating hinge 6, the lamp bead 8 is fixedly connected with the spotlight shell 7 and is positioned in the spotlight shell 7, and the lens 9 is fixedly connected with the spotlight shell 7 and is positioned on one side of the spotlight shell 7.
The supporting device body further comprises a glass sheet 15, wherein the glass sheet 15 is detachably connected with the shell 2 and arranged below the reflecting mirror 1 and the lighting module, and the glass sheet is used for placing an observed sample.
The housing 2 has a dimmer switch 16, the dimmer switch 16 being located on a side of the housing 2.
In this embodiment, the supporting device plays a role in supporting the reflecting mirror 1, so as to fix and adjust the position and the reflecting angle of the reflecting mirror. The shooting can be performed by using natural light or a light source of a microscope, and a user selects a proper light source according to actual conditions. In order to enable better shooting effect, the spotlight assembly is used for spotlight, and the housing 2 is used for reducing influence of ambient light. The shell 2 provides mounting conditions for the lighting module, reduces the influence of natural light, realizes a better light condensation effect with the lighting module, and improves the definition of an observation sample; the spotlight shell 7 is made of metal materials, the lens 9 is a TIR lens 9, the spotlight angle of the lens 9 is smaller than 30 degrees (for example, 30 degrees or 20 degrees), the smaller the angle is, the better the spotlight effect can be achieved, the better the effect of intensively illuminating samples can be achieved, the spotlight shell 7 and the spotlight base 10 are made of aluminum shells for radiating the lamp beads 8, meanwhile, the spotlight shell 7 and the lamp beads 8 are prevented from being damaged due to overhigh temperature in use, fine adjustment of the illumination angle of a light source is achieved at the position of the spotlight rotating hinge 6, the lamp beads 8 are made of lamp beads 8 with the intensity being adjusted by the aid of a dimming switch (the lamp beads 8 can be made of light emitting diodes, laser diodes, semiconductor laser tubes and the like), and a USB switch, a battery box or a 220V power supply can be connected to supply power for devices.
The supporting device body is used for fixing and adjusting the angle of the reflecting mirror 1. The reflector 1 is fixed by adopting the supporting device body and then is placed on the objective table, so that the side surface of the cell sphere can be observed. According to the invention, the microscope at the bottom of the sample can be observed by adding the refraction of the reflector 1, so that the side surface of the sample can be observed, and the observation of multiple angles of the sample is realized. When a sample is required to be photographed at multiple angles, the reflecting mirror 1 is placed above the microscope objective, the sample is placed on the front side (namely, the surface with specular reflection) of the reflecting mirror 1, the angle and the sample position of the reflecting mirror 1 are adjusted, the observation of the side surface of the cell ball is realized through the reflection relation, the image of the side surface of the sample is captured by the objective after being reflected by the reflecting mirror 1 through the relative position of a microscope platform moving device and the microscope objective, the side surface of the sample is observed, the observation of the same sample in different dimensions is realized, the bottom surface and the side surface of the sample are photographed by using the monitoring device, and special surface treatment is not required, so that the cost control to a large extent is realized, and the technical difficulty of preparing the sample for multi-angle observation is reduced; the sample is not required to be contacted, so that the sample pollution is reduced to the greatest extent; meanwhile, expensive facility equipment is not needed, the threshold for observing the samples at multiple angles is reduced, shooting can be carried out through a laboratory conventional microscope, and the problem that the conventional microscope cannot realize observation of the samples at multiple angles on the microscope is solved.
Example 1:
the three-dimensional cell sphere side and bottom surface bright field images were photographed using an inverted fluorescence microscope in combination with the microscope sample multi-angle photographing assembly of the first embodiment.
The cell pellet is transferred to the liquid sample tank 17. Specifically, the liquid sample tank 17 includes a transparent casing 18 and an optical glass sheet 19, and the liquid sample tank 17 is used for Cheng Fanghuan flushing liquid and cell balls, and other sample tanks meeting the requirements of containing samples can be used instead.
In order to achieve a better shooting effect, the cell ball is adjusted to be close to one side of the liquid sample groove 17 close to the optical glass sheet 19;
placing the glass surface of the liquid sample tank 17 in front of the reflecting mirror 1 and ensuring that the reflecting mirror can display the image of the photographed surface of the cell ball;
opening a microscope, observing the state in an eyepiece of the microscope, adjusting the lighting module to a proper position, adjusting the spotlight to a proper brightness through the dimming switch 16, finding an image of a sample in the reflector 1 in a larger visual field, adjusting the angle of the spotlight to achieve a better shooting effect, and sequentially adjusting the coarse focusing spiral and the fine focusing spiral to obtain a clear image;
switching different objective lenses to shoot multiple sample side images;
finding an image of the bottom of the sample in a larger visual field, adjusting the spotlight to a proper brightness and angle through the dimming switch 16, and sequentially adjusting the coarse focusing spiral and the fine focusing spiral to obtain a clear image;
and switching different objective lenses to perform multiple image shooting of the bottom of the sample.
Example 2:
the three-dimensional cell sphere side and bottom surface bright field and dio+dapi stained fluorescence images were photographed using an inverted fluorescence microscope in combination with the components of the microscope of the first embodiment photographed at multiple angles with the sample.
Transferring the stained cell ball into the liquid sample groove 17 in a dark place, filling the liquid sample groove 17 with buffer solution to achieve the best shooting effect, and adjusting the sample to the side, which is close to the liquid sample groove 17 and is close to the optical glass sheet 19;
placing the glass surface of the liquid sample tank 17 in front of the reflecting mirror 1 and ensuring that the reflecting mirror can display the image of the photographed surface of the cell ball;
turning on the microscope, adjusting the spotlight to a proper brightness through the dimming switch 16, finding the image of the sample in the reflector 1 in a larger visual field, and adjusting the spotlight angle to achieve a better shooting effect;
turning off the spotlight, turning on a self-contained fluorescent light source of the microscope, adjusting the light source to proper intensity, and sequentially adjusting the coarse focusing spiral and the fine focusing spiral to obtain a clear image;
switching different objective lenses to shoot multiple sample side fluorescent images;
finding an image of the bottom of the sample in a larger field of view, dimming the spot light to a suitable brightness by means of the dimmer switch 16;
turning off the shot lamp, turning on a self-contained fluorescent light source of the microscope, adjusting the light source to proper intensity, and sequentially adjusting the coarse focusing spiral and the fine focusing spiral to obtain clear images;
and switching different objective lenses to shoot multiple fluorescent images of the bottom surface of the sample.
Example 3:
the inverted fluorescence microscope was used in combination with the sample multi-angle photographing assembly of the first embodiment to conduct droplet contact angle photographing, to characterize hydrophilic/hydrophobic properties, and surface energy properties of the material.
Specifically, samples to be tested, such as acrylic acid ester, polydimethylsiloxane, polypropylene and Parafilm films, were placed in the sample tank 17, and 4. Mu.L of water was dropped onto the samples with a microinjector to form droplets.
Placing the glass surface of the liquid sample tank 17 in front of the reflecting mirror 1 and ensuring that the reflecting mirror can display the image of the photographed surface of the liquid drop;
the microscope is opened, the state in the eyepiece of the microscope is observed, the lighting module is adjusted to a proper position, the spotlight is adjusted to a proper brightness through the dimming switch 16, the image of the sample in the reflector 1 is found in a larger visual field, the angle of the spotlight is adjusted to achieve a better shooting effect, and the coarse focusing spiral and the fine focusing spiral are sequentially adjusted to obtain a clear image (please refer to fig. 10A). The difference in contact angle measurement between the results obtained in this example and the professional contact angle measuring instrument is negligible (please refer to fig. 10B), which shows that it can be used as an economical self-made device to evaluate surface wettability and surface energy.
Example 4:
the underwater superoleophobic properties of the paper-like material were evaluated by underwater monitoring the oil drop contact angle of the material surface using an inverted fluorescence microscope in combination with the assembly of the microscope in the first embodiment photographed at multiple angles with the sample.
Specifically, a sample to be measured, such as filter paper, printing paper, cardboard, is placed in the sample tank 17, and 2. Mu.L of a red oily liquid is added to the sample with a microinjector.
Placing the glass surface of the liquid sample tank 17 in front of the reflecting mirror 1 and ensuring that the reflecting mirror can display the image of the photographed surface of the liquid drop; water is then added to sample tank 17 to submerge the paper.
The microscope is opened, the state in the eyepiece of the microscope is observed, the lighting module is adjusted to a proper position, the spotlight is adjusted to a proper brightness through the dimming switch 16, the image of the sample in the reflector 1 is found in a larger visual field, the angle of the spotlight is adjusted to achieve a better shooting effect, the coarse focusing spiral and the fine focusing spiral are sequentially adjusted to obtain a clear image, and the process that red oily fuel is separated out from the surface of paper in an underwater environment to form oil drops is observed (please refer to FIG. 11). The results obtained by this example were used to analyze the underwater oleophobic properties of the material.
Example 5:
the hydrogen bubble density on the electrode surface was monitored using an inverted fluorescence microscope in combination with the sample multi-angle photographing assembly of the microscope in the first embodiment, and the hydrogen production efficiency by water electrolysis was evaluated.
Specifically, a three-electrode reaction system (working electrode, counter electrode, reference electrode) in the electrolytic water reaction is placed in the sample tank 17, and the electrodes are communicated with an electrochemical workstation through wires.
An electrolyte, for example, a 1mol/L KOH solution, is added to the three-electrode reaction system.
Placing the glass surface of the liquid sample cell 17 in front of the mirror 1 and ensuring that the mirror is able to observe the phenomenon of the working electrode surface; a voltage, for example-1.8V, is then applied through the electrochemical workstation.
The angle of the spotlight is adjusted to achieve a good shooting effect, the coarse focusing spiral and the fine focusing spiral are sequentially adjusted to obtain clear images, and the process that bubbles are separated out from the surface of the working electrode and released in the electrolyte is observed (please refer to fig. 12).
This example enables in situ monitoring of bubble formation dynamics, thus enabling cost-effective rapid screening and evaluation of the effect of catalyst catalytic efficiency in clean energy sources.
Second embodiment:
the reflection mirror comprises a supporting device body and a reflection mirror 1, wherein the reflection mirror 1 is movably connected with the supporting device body and is positioned on one side of the supporting device body.
The support device body comprises a shell 2 and a lighting module, the shell 2 is provided with a sample placing groove 3, the lighting module is arranged on the inner side wall of the sample placing groove 3, and the reflecting mirror 1 is movably connected with the inner side wall of the sample placing groove 3.
The lighting module comprises a spotlight fixing base 5, a spotlight rotating hinge 6, a spotlight shell 7, a lamp bead 8 and a lens 9, wherein the spotlight fixing base 5 is fixedly connected with the inner side wall of the sample placing groove 3, the spotlight rotating hinge 6 is fixedly connected with the spotlight fixing base 5 and is positioned on one side of the spotlight fixing base 5, the spotlight shell 7 is fixedly connected with the spotlight rotating hinge 6 and is positioned on one side, far away from the spotlight fixing base 5, of the spotlight rotating hinge 6, the lamp bead 8 is fixedly connected with the spotlight shell 7 and is positioned in the spotlight shell 7, and the lens 9 is fixedly connected with the spotlight shell 7 and is positioned on one side of the spotlight shell 7.
The shell 2 is provided with an objective shooting port 4, and the objective shooting port 4 is communicated with the sample placing groove 3 and is positioned above the reflecting mirror 1.
The housing has a dimmer switch 16, the dimmer switch 16 being located on a side of the housing.
In this embodiment, unlike the first example, with respect to the front microscope, it is possible to observe and photograph a sample more conveniently through the objective photographing port 4. The shell 2 provides mounting conditions for the lighting module, reduces the influence of natural light, and then realizes a better light condensing effect by matching with the lighting module, so that the definition of an observation sample is further improved; the spotlight shell 7 is made of metal materials, the lens 9 is a TIR lens 9, the spotlight angle of the lens 9 is smaller than 30 degrees (for example, 30 degrees or 20 degrees), the smaller the angle is, the better the spotlight effect can be achieved, the better the effect of intensively illuminating samples can be achieved, the spotlight shell 7 and the spotlight base 10 are made of aluminum shells for radiating the lamp beads 8, meanwhile, the spotlight shell 7 and the lamp beads 8 are prevented from being damaged due to overhigh temperature in use, fine adjustment of the illumination angle of a light source is achieved at the position of the spotlight rotating hinge 6, the lamp beads 8 are made of lamp beads 8 with the intensity being adjusted by the aid of a dimming switch (the lamp beads 8 can be made of light emitting diodes, laser diodes, semiconductor laser tubes and the like), and a USB switch, a battery box or a 220V power supply can be connected to supply power for devices.
Examples:
the structural differences of the stained artificial sweat-stained artificial sebum mixture separating layer, and the stained secondary water-stained artificial sebum mixture separating layer were observed using an upright fluorescence microscope in combination with the second embodiment.
Specifically, weighing 0.9g of triglyceride, 0.34g of oleic acid, 0.26g of squalene and 0.5g of jojoba oil, adding into a centrifuge tube, and ultrasonically mixing for 5min to obtain artificial sebum;
adding 1mg of oil red powder into 50 mu L of artificial sebum, and ultrasonically mixing for 10min until no obvious oil red particles exist in the mixed solution to obtain dyed artificial sebum;
taking 100mL of artificial sweat, and adding 5 mu L of green aqueous dye stock solution to obtain dyed artificial sweat;
adding 50 mu L of dyed artificial sebum and 60wt% of emulsifier SDS into 100mL of dyed artificial sweat, and uniformly mixing by ultrasonic for 30min to obtain dyed artificial sweat-dyed artificial sebum mixed solution;
taking 100mL of secondary water, and adding 5 mu L of green aqueous dye stock solution to obtain dyed secondary water;
adding 50 mu L of dyed artificial sebum and 60wt% of emulsifier SDS into 100mL of dyed secondary water, and uniformly mixing by ultrasonic for 30min to obtain dyed secondary water-dyed artificial sebum mixed solution;
respectively taking 100 mu L of dyed artificial sweat-dyed artificial sebum mixed solution and dyed secondary water-dyed artificial sebum mixed solution, adding into different centrifuge tubes, and centrifuging at 3000rpm for 10min;
after centrifugation, lightly blowing layered liquid in the centrifuge tube uniformly for 3-5 times by using a pipette, and transferring the layered liquid into the liquid sample tank 17;
different objective lenses were switched, multiple-stained artificial sweat-stained artificial sebum mixture separation layer shots were performed, and multiple-stained secondary water-stained artificial sebum mixture separation layer shots were performed.
Third embodiment:
the support device body comprises a base 10, a reflector support frame 11, a rotary fixing support 12 and a reflector fixing table 13, wherein the base 10 is fixedly connected with one end of the reflector support frame 11, the other end of the reflector support frame 11 is provided with a rotary shaft, one end of the rotary fixing support 12 is movably connected with the rotary shaft, the reflector fixing table 13 is movably connected with the other end of the rotary shaft fixing frame, the reflector 1 is fixedly connected with the side face of the reflector fixing table 13, and the reflector fixing table 13 is further provided with a rotary angle observation hole 14.
In this embodiment, the base 10 is used for maintaining the gravity center of the device, and a material with a higher density is selected for manufacturing (metal) or adding a balancing weight; the side edges of the base 10 and the reflector support frame 11, which lean against the reflector 1, do not exceed the reflector 1, otherwise, the reflector 1 cannot flexibly adjust the shooting angle; the rotary fixing bracket 12 not only can adjust the angle of the reflecting mirror 1 between 0 degrees and 90 degrees, but also can adjust the height of the reflecting mirror 1 relative to the platform according to different samples, thereby meeting the shooting requirements of samples in different shapes and different angles; the current angle of the reflector 1 and the height position of the relative platform can be measured through the positions of the observation rotation angle observation hole 14 and the rotation fixing bracket 12; the mirror 1 is flush with the mirror fixing table 13 when being connected to the base 10, so that the mirror 1 can be adjusted at an angle of 0-90 degrees and the relative height can be adjusted. Examples:
the inverted microscope was used in combination with the assembly of the microscope of the third embodiment for multi-angle photographing of samples to photograph hot melt deposition to print different angle side structures.
Sample preparation process: modeling a sample with the height of 1mm and the length and width of 5cm by using SOLIWORKS software; exporting the file in a stl file, loading the file into Cura software for slicing, and setting a printing material as a PLA material; setting the temperature of a nozzle to 200 ℃, setting the temperature of a printing hot table to 55 ℃, enabling the height of an initial layer to be 0.3mm, enabling the moving speed of a spray head to be 100mm/s, enabling the thickness of a printing layer to be 0.1mm, enabling the thickness of the printing bottom to be 1mm, selecting the diameter of a printing line material to be 1.75mm, and selecting unsupported printing; the file is saved as a gcode format and exported to a Sen K6 printer, and the printer is leveled by a printing platform after being started; and selecting a printing file, and waiting for the printing platform to cool to room temperature after printing is finished.
Taking down the sample from the printing platform, selecting the most straight side to be placed below the reflecting mirror 1, and adjusting the angle and the relative height of the reflecting mirror 1 to the position capable of observing the side surface to be observed of the sample through the rotating shaft and the reflecting mirror fixing table 13 shaft;
sequentially adjusting a coarse focusing spiral and a fine focusing spiral, and adjusting the image of the side surface of the sample in the view field internal reflector 1 to be clear;
and switching different objective lenses to shoot multiple fluorescent images of the bottom surface of the sample.
Referring to fig. 14, a fourth embodiment provides a multi-angle photographing system for a sample for a microscope, which includes any one of the first, second and third embodiments, the multi-angle photographing assembly for a sample for a microscope,
and the microscope objective lens is included, and the included angle between the microscope objective lens and the reflecting mirror 1 is between 0 and 90 degrees, and is not equal to 0 or 90 degrees.
In this embodiment, when light is irradiated to a smooth reflecting surface according to the law of reflection of light, the reflection of light is called specular reflection, and the law of reflection is followed by the reflection of light: 1. the reflected light, the incident light and the reflecting surface are positioned on the same plane; 2. the angle of reflection is equal to the angle of incidence; 3. the reflected light and the incident light are on opposite sides of the normal. It is known from the law of reflection that when a light ray from an object irradiates the mirror surface of the plane mirror 1, the light ray will be reflected by the angle identical to the incident angle, and a virtual image of the object is formed in the mirror surface and is in mirror image relationship with the object. And placing the sample above a microscope platform, and adjusting the observation hole of the microscope platform, the objective lens, the reflector 1 and the position of the sample to be observed. When observing the bottom surface of the sample, the bottom surface of the sample is required to be positioned right above the microscope objective lens, so that the image of the bottom of the sample is captured by the objective lens; when observing the side of the sample, the sample needs to be adjusted to enable the observation surface to be parallel to the reflecting surface of the reflecting mirror 1, so that the observed image can be better captured by the microscope objective lens. The condition that the image of the sample can be captured by the microscope objective is that the surface of the sample to be observed is in a symmetrical relationship with the microscope objective with respect to the normal of the mirror surface of the mirror 1. Since the microscope objective cannot be moved at will, the mirror 1 and the sample position can be moved accordingly, so that an image of the sample is captured by the microscope objective through the mirror 1. The angle between the optical path of the image of the sample reflected by the mirror 1 and the normal to the mirror 1 should be between 0 deg. and 90 deg., and not equal to 0 deg. or 90 deg.. When the included angle between the light path and the normal line of the reflector 1 is between 0 and 45 degrees, the image of the upper side surface of the sample can be observed through reflection; when the included angle between the light path and the normal line of the reflector 1 is between 45 and 90 degrees, the image of the lower side surface of the sample can be observed through reflection; when the angle between the light path and the normal of the reflector 1 is 45 degrees, the image of the front side of the sample can be observed through the reflector 1. Thereby realizing multi-angle shooting of the sample.
Referring to fig. 11 to 12, a fifth embodiment provides a multi-angle photographing method for a sample for a microscope, which is applied to any one of the first, second and third embodiments of the multi-angle photographing assembly for a sample for a microscope, and includes the following steps:
s1, sequentially reflecting images of different surfaces of a sample through the reflecting mirror 1;
s2, capturing images of the reflector 1 through a microscope in sequence, and shooting.
Specifically, according to shooting requirements, the observed surface of the sample is placed in front of the reflecting mirror 1, and the positions and angles among the supporting device, the sample and the microscope are adjusted, so that the microscope can observe images of different surfaces of the sample through the reflecting mirror 1, and then shooting is carried out, thereby realizing multi-angle shooting of the sample.
The above disclosure is merely illustrative of preferred embodiments of the multi-angle photographing assembly, photographing system and photographing method for a microscope sample, and it is needless to say that the scope of the invention is not limited thereto, and those skilled in the art will understand that all or part of the procedures for implementing the embodiments are performed and equivalent changes are included in the scope of the invention.
Claims (9)
1. An assembly for multi-angle photographing of a sample for a microscope, characterized in that,
the device comprises a supporting device body and a reflecting mirror, wherein the reflecting mirror is connected with one side of the supporting device body.
2. The multi-angle imaging module for a microscope according to claim 1, wherein,
the supporting device body comprises a shell and a lighting module, the shell is provided with a sample placing groove, and the lighting module and the reflecting mirror are connected with the inner side wall of the sample placing groove.
3. The multi-angle imaging module for a microscope according to claim 2, wherein,
the shell is provided with an objective shooting port, and the objective shooting port is communicated with the sample placing groove and is positioned above the reflecting mirror.
4. The multi-angle imaging module for a microscope according to claim 3, wherein,
the lighting module comprises a spotlight fixing base, a spotlight rotating hinge, a spotlight shell, a lamp bead and a lens, wherein the spotlight fixing base is fixedly connected with the inner side wall of the sample placing groove, the spotlight rotating hinge is fixedly connected with the spotlight fixing base and is located on one side of the spotlight fixing base, the spotlight shell is fixedly connected with the spotlight rotating hinge and is located on one side of the spotlight fixing base, the spotlight bead is fixedly connected with the spotlight shell and is located in the spotlight shell, and the lens is fixedly connected with the spotlight shell and is located on one side of the spotlight shell.
5. The multi-angle imaging module for a microscope according to claim 4, wherein,
the supporting device body further comprises a glass sheet, wherein the glass sheet is detachably connected with the shell, and is arranged below the reflecting mirror and the lighting module.
6. The multi-angle imaging module for a microscope according to claim 5, wherein,
the housing has a dimmer switch disposed thereon.
7. The multi-angle imaging module for a microscope according to claim 1, wherein,
the supporting device comprises a supporting device body, and is characterized by further comprising a base, a reflector supporting frame, a rotary fixing support and a reflector fixing table, wherein the base is fixedly connected with one end of the reflector supporting frame, a rotating shaft is arranged at the other end of the reflector supporting frame, one end of the rotary fixing support is movably connected with the rotating shaft, the reflector fixing table is movably connected with the other end of the rotating shaft fixing frame, the reflector is fixedly connected with the side face of the reflector fixing table, and the reflector fixing table is further provided with a rotating angle observation hole.
8. A multi-angle photographing system for a sample for a microscope, comprising the multi-angle photographing assembly for a sample for a microscope according to any one of claims 1 to 7, characterized in that,
the microscope comprises a reflecting mirror, and the included angle between the microscope objective and the reflecting mirror is between 0 and 90 degrees, and is not equal to 0 or 90 degrees.
9. A method for photographing a sample for a microscope at multiple angles, applied to the assembly for photographing a sample for a microscope according to any one of claims 1 to 7, comprising the steps of:
sequentially reflecting images of different surfaces of the sample by the reflecting mirror;
the images in the reflecting mirror are captured sequentially by a microscope, and shooting is performed.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311276350.6A CN117270186A (en) | 2023-09-30 | 2023-09-30 | Multi-angle shooting assembly, shooting system and shooting method for microscope sample |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202311276350.6A CN117270186A (en) | 2023-09-30 | 2023-09-30 | Multi-angle shooting assembly, shooting system and shooting method for microscope sample |
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| CN117270186A true CN117270186A (en) | 2023-12-22 |
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- 2023-09-30 CN CN202311276350.6A patent/CN117270186A/en active Pending
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