CN104568859A - Fluorescence observation device, base frame and fluorescence microscope with multiple groups of different-angle light sources - Google Patents

Abstract

The utility model provides a fluorescence observation device, bed frame and fluorescence microscope, wherein fluorescence observation device includes the cover body, multi-angle light-emitting unit and control unit, and the cover body includes printing opacity heat insulation layer and heat dissipation layer, and multi-angle light-emitting unit has a plurality of narrow light field exciting light components, can set up narrow light field exciting light component heat conduction on the heat dissipation layer for the heat dissipation, the light beam that narrow light field exciting light component with directive property sends out is then passed through printing opacity heat insulation layer, and control unit control multiunit different narrow light field exciting light component of angle of giving out light, in order to select the angle of illumination, fluorescence observation device can combine into the bed frame with a base, narrow light field exciting light component also can locate on the base, by in order to select the excitation light angle of illumination, construct the required environment of.

Description

Fluorescence observation device, base frame and fluorescence microscope with multiple groups of different-angle light sources

【Technical field】

The invention relates to a fluorescence observation device, in particular to a fluorescence observation device applicable to a fluorescence micro-optic device, a base frame for the fluorescence micro-optic device and the device.

【Background technique】

In addition to being used in industrial inspection, counterfeit banknote identification, and criminal identification, fluorescence technology has been extended to the analysis and tracking of genetic engineering and gene transplantation experiments in biological research in recent years, which has gradually increased the importance of fluorescence image identification and analysis.

The so-called fluorescence here is mainly to irradiate a high-frequency light on the object to be observed with fluorescence characteristics, such as the anti-counterfeiting thread of banknotes, or the location of suspected bloodstains at the crime scene, so as to excite a lower-frequency fluorescence. Coupled with an appropriate filter combination, the fluorescent images of the above-mentioned banknotes or bloodstains can be clearly viewed or captured; as for many studies in the field of biological sciences, they are aimed at gene transfer, and the implanted genes often produce fluorescence for easy observation The success or failure of gene implantation can be confirmed by the presence or absence of the fluorescent reaction of the object to be observed, and further in-depth research can be carried out on the organisms with successful transplantation.

At present, the common fluorescent animals include white mice, maggots, various fishes, jellyfish, etc. Because various experimental animals are quite different from each other regardless of their size or the wavelength of excitation light that stimulates fluorescence. For animals that are slightly larger and can be observed with the naked eye, it is usually difficult to limit the range of motion. How to limit the range of motion and provide economical and easy-to-operate appropriate auxiliary equipment for observation is a homework that must be completed before the experiment. Even if the experimental white mice are usually fed in the animal center, due to the strong fecundity and activity of the white mice, it is necessary to identify and confirm that the experimental white mice do have fluorescence reactions from the moment they are picked up, and a light and easy-to-carry fluorescence observation device is needed. .

In recent years, there is a fluorescent gene zebrafish that has been extensively studied, mainly because this type of zebrafish has a large reproductive capacity, a short reproductive cycle, can produce a large number of offspring, and has characteristics similar to human organ systems, and has even been extended to disease research, Drug screening, toxicology testing and other related research. However, if the experimental animal is a zebrafish the size of a sesame seed, it is necessary to use a microscope with different magnifications to observe and record. Fluorescence microscopes commonly used at present, although they are convenient to use, can effectively assist researchers, for example, in observing analytes with fluorescence reactions; More attention is paid to how to restrict the traveling route of the excitation beam in order to theoretically increase the resolution and reduce the error of the image, and its structure does not deviate from the main optical design of the previous microscope.

However, when the excitation beam travels a long distance, the energy diffuses inversely proportional to the square of the distance. In order to solve the problem of insufficient energy when the excitation light reaches the object to be tested, the existing fluorescence microscope has greatly increased the brightness of the light source, causing some small objects to be tested, such as zebrafish, to denature due to excessive heating during the observation process. In addition, since the excitation light irradiates the surface of the object to be observed in a vertical direction, the direct reflection light is very strong, and the intensity of the fluorescence released after absorbing the excitation light is much lower than that of the direct reflection light. In other words, as noise, the intensity of reflected light with a shorter wavelength is thousands of times stronger than the fluorescent signal with a longer wavelength that is actually to be observed and recorded. The signal-to-noise (S/N) ratio is too poor, resulting in extremely complicated configuration Expensive optical structures also make fluorescence microscopes cost millions.

In particular, the existing fluorescence microscopes are not compatible with ordinary optical microscopes, and it is also forced in a disguised form to require those who observe fluorescence reactions to abandon the existing microscopes in the research room and re-purchase expensive fluorescence microscopes. This does not meet general expectations, especially the price of the microscope and its performance. The correlation is very high. The number of lenses and the quality of the manufacturing process are enough to greatly affect its price. If you have already purchased a general optical microscope with high performance, it is even more impossible to easily purchase another fluorescence microscope with the same high performance. Therefore, how to easily convert a general optical microscope into a fluorescent microscope capable of locking fluorescence characteristics by adding an auxiliary device has also become the focus of further research in the present invention.

In order to facilitate analysis and observation, current fluorescence-related biological experiments often use fluorescent proteins that are excited by ultraviolet light, deep blue light or green light to release blue light, green light or red light. An existing assisting device 70 for overcoming difficulties is shown in FIG. 10 . For example, an ultraviolet light source 72 is directly arranged at the front ends of two bellows tubes 74 , and the user is allowed to bend the bellows tubes 74 to pick up an appropriate light angle and irradiate to the costumes. There is a petri dish 76 with a fluorescent object to be observed, so as to generate photoluminescence, but limited by the structure of the bellows 74, when the distance between the objective lens 78 of the optical microscope and the target object is only two or three centimeters, the bellows 74 will be difficult to get from such a narrow Turning into the space. Not only the angle cannot be adjusted precisely, so the light angle is often not as expected, but also the brightness in the light range is not easy to be uniform, so that the captured images cannot be accurately and quantitatively analyzed.

In addition, experimental animals such as zebrafish are still living in the experiment, so when the above-mentioned fluorescence microscopy experiment is carried out, when the object to be observed is, for example, fluorescent zebrafish or maggots, the target object may move continuously, not to mention moving in small water droplets. The zebrafish may also die in the experiment due to insufficient dissolved oxygen in the water droplets. The operating time and the influence of the experimental environment on the organism must be considered at the same time. The uneven small-scale illumination is even more annoying. If the operation is not skilled or unlucky Good, it is easy to lead to failed experimental results. Larger animals, such as white mice, cannot be ordered to stay in the range of the light source, and they wander around and eat, sometimes causing damage and pollution to experimental equipment.

In order to enable the above-mentioned living organisms to obtain a good lighting effect, the applicant of the present invention has first applied in the 201210402931.5 application "Fluorescence Auxiliary Module for Microscopic Optical Pickup Device and the Microscopic Optical Pickup Device", as shown in Figure 11 As shown, the predetermined observation position of the base 84 is covered by the cover 82, and a light-shielding chamber 86 can be formed together with the base 84, and the object to be observed is arranged in the light-shielding chamber 86 for Observe, and a plurality of light source assemblies 88 are arranged on the inner wall of the shield 82 , and the light source assemblies 88 are fine-tuned along a track by the adjusting member 80 to move on the surface of the shield 82 to provide a better lighting angle.

Although the above direction can make the illumination uniform and increase the convenience of observation, if the heat generated by the light source assembly does not have a good heat dissipation pipe, so that when the heat is left in the light-shielding chamber, the temperature may still rise, resulting in the final The experimental results are not good, and in order to obtain the best lighting angle, it is necessary to design an additional mechanical or electronic adjustment structure. It is difficult to reduce the development cost, and it is prone to damage after long-term use. Additional maintenance is required, so there is still room for improvement. .

Therefore, a fluorescence observation device with uniform illumination, easily adjustable luminous angle, and convenient and rapid overall operation is selected, so that researchers do not need to be distracted to operate the light source during the observation and recording process, and properly shield external interference light, so that the experimental process It is smooth and easy to observe and record. At the same time, it increases the heat dissipation effect, so that the heat generated by the light is not easy to stay in the mask, slows down the temperature rise, and simplifies the structure of the light angle adjustment, reduces the production cost and improves the service life of the product.

To sum up, the present invention provides a more excellent fluorescence observation device and micro-optical acquisition device. On the one hand, when the experimental animals are large, a simple fluorescence observation device can be selected for direct observation and recording. Relatively, when the experimental animal is small in size, it can also be equipped with a more sophisticated fluorescence observation device or combined with the microscope base frame, so that the general microscope or micro camera can be easily upgraded to meet the needs of fluorescence research; On the one hand, while simplifying the structure and improving the convenience of operation, the quality of microscopic observation records will not be sacrificed, and a very stable and simple operation platform is proposed to meet various experimental application requirements, and at the same time, it has an extremely economical price. Advantages, so that it has an unparalleled cost performance.

The present invention is completely different from the design of the previous fluorescence microscope. It adopts the proximity light source structure and designs a proximity excitation light source with selectable angles, so as to obtain dark-field fluorescence images. And by means of dark field lighting, the interference of reflected light can be eliminated, so that the intensity of incident light can be greatly reduced, and the image quality can still be ensured; on the other hand, the brightness of the light source can be greatly reduced again by using the proximity light source, so that the LED can be smoothly introduced and placed on the object to be observed The surrounding area shortens the path of the incident light, especially has a layer of light-transmitting and heat-insulating inner shell, which avoids unnecessary increase in the temperature in the observed area, and also prevents larger observed animals from touching or polluting the environment. Damaged light source. The structural design is simplified, the manufacturing cost is greatly reduced, and it can even be used as an auxiliary device alone, or it can be used in conjunction with the existing optical microscope operation. Through the disclosure of the present invention, researchers can obtain products compatible with existing equipment at an extremely economical price, greatly improving research quality.

【Content of invention】

An object of the present invention is to provide a fluorescence observation device with a modular structure, which simplifies the operation of research and observation, forms a stable and friendly operation platform, and further improves the success rate of research experiments.

Another object of the present invention is to provide a fluorescence observation device for isolating the heat generated by the light source assembly, so as to prevent the living organism under test from directly touching the light source assembly, or scalding or scorching the animal under test due to the transfer of heat energy into the observation environment.

Another object of the present invention is to provide a fluorescence observation device that uses a light-transmitting heat-insulating layer to isolate experimental animals from contacting light source components, so as to avoid damage and contamination of experimental animals to the light source.

Another object of the present invention is to provide a pedestal with a fluorescence observation device. Since the low-angle excitation light source has multiple groups of light-emitting elements with different light-emitting angles, the structure is simplified, and the fluorescence observation device with adjustable light-emitting direction reduces the production cost. cost and increase product lifespan.

Another object of the present invention is to provide a pedestal with the above-mentioned fluorescence observation device, which can be easily connected due to its high compatibility with existing general optical microscopes or microscopic cameras and other microscopic optical pickup devices, thus improving the It can be used from optical microscope to fluorescence, increasing the application flexibility of general microscopic optical pickup devices.

Yet another object of the present invention is to provide a base frame for a fluorescence micro-optical device with a mask, which can shield the interference of external strong light and stray light, and improve the quality of fluorescence microscopic imaging.

Still another object of the present invention is to provide a fluorescence microscope using the above-mentioned fluorescence observation device. By providing a close-to-light source, the luminous intensity is reduced, the accompanying unnecessary heat energy is reduced, and the heat generated by the light source is prevented from entering the shield needlessly. Furthermore, the heat dissipation effect of the mask is increased, so that the temperature of the environment where the object to be observed is located is not easy to rise rapidly.

Yet another object of the present invention is to provide a fluorescence microscope with heat dissipation effect, so that the heat energy emitted by the light source during energy conversion is guided away, reducing the interference of the object to be observed when the temperature rises, and improving the efficiency and accuracy of the experiment.

A fluorescence observation device with multiple sets of different-angle light sources of the present invention is provided for a cover to be placed in a load-bearing observation area for observing a fluorescence object to be observed located in the load-bearing observation area, wherein the fluorescence observation device includes: a cover body, An accommodating space for accommodating the fluorescent object to be observed is surrounded together with the above-mentioned bearing observation area, and an observation hole is formed in the cover body, and the cover body includes: a layer of light-transmitting and heat-insulating layer facing the direction of the accommodating space setting; and a layer of heat dissipation layer, the thermal conductivity of which is higher than that of the above-mentioned light-transmitting heat-insulating layer, and the light transmittance in at least the range of visible light is lower than that of the above-mentioned light-transmitting heat-insulating layer. The layer is far away from the direction of the accommodation space; the above-mentioned fluorescence observation device with multiple groups of different-angle light sources also includes a group of multi-angle light-emitting units with a plurality of narrow light field excitation light elements, each of the aforementioned narrow light field excitation light elements emits The light beams pass through the above-mentioned light-transmitting and heat-insulating layer respectively, and irradiate to the accommodating space, and the above-mentioned narrow light field exciting light element has at least a plurality of different irradiation angles from each other; and a control unit is used to control at least one of the multi-angle light emitting units This angle of illumination of the narrow light field excitation light element described above.

A pedestal for a fluorescent micro-optical device of the present invention is used for observing a fluorescent object to be observed, and the pedestal includes: a base with a carrying observation area for observing the fluorescent object to be observed; a cover , together with the above-mentioned base, an accommodating space for accommodating the fluorescent object to be observed is formed, and the cover body is formed with an observation hole, and the cover body includes: a layer of light-transmitting and heat-insulating layer facing the direction of the accommodating space setting; and a layer of heat dissipation layer, the thermal conductivity of which is higher than that of the above-mentioned light-transmitting heat-insulating layer, and the light transmittance in at least the range of visible light is lower than that of the above-mentioned light-transmitting heat-insulating layer. The layer is away from the direction of the accommodation space; the base frame also includes a group of multi-angle light-emitting units with a plurality of narrow light field excitation light elements, and the light beams emitted by each of the aforementioned narrow light field excitation light elements respectively travel through the above-mentioned light-transmitting and heat-insulating layer , irradiate to the accommodating space, and the aforementioned narrow light field excitation light element has at least a plurality of mutually different irradiation angles; and a control unit is used to control at least the above narrow light field excitation light element of the multi-angle light emitting unit The angle of illumination.

A fluorescent micro-optical device of the present invention is for observing a fluorescent object to be observed, comprising: a body; an objective lens mounted on the body; and a fluorescence observation device, comprising: a base with a A bearing observation area for observing the fluorescent object to be observed; a cover body, together with the above-mentioned base, forms an accommodating space for accommodating the fluorescent object to be observed, and the cover body is formed with an observation hole for the lens lens The image of the object to be observed located in the accommodating space is captured through the observation hole, and the cover body includes: a layer of light-transmitting heat-insulating layer, which is arranged facing the direction of the accommodating space; and a heat dissipation layer, whose thermal conductivity is higher than The above-mentioned light-transmitting heat-insulating layer, and at least the light transmittance in the visible light range is lower than the above-mentioned light-transmitting heat-insulating layer, and the heat dissipation layer is arranged on one side of the light-transmitting heat-insulating layer away from the accommodation space; the fluorescence observation device also It includes a group of multi-angle light-emitting units with a plurality of narrow light field excitation light elements, and the light beam emitted by each of the aforementioned narrow light field excitation light elements respectively travels through the above-mentioned light-transmitting and heat-insulating layer, and irradiates to the accommodating space, and the aforementioned narrow light field The excitation light element has at least a plurality of mutually different illumination angles; and a control unit is used for controlling the illumination angle of at least the narrow light field excitation light element of the multi-angle light-emitting unit.

Therefore, the fluorescent observation device of the present invention, the pedestal for the fluorescent micro-optical device, and the device utilize the arc curve of the cover body so that the narrow light field excitation light elements arranged on the cover body or the base can respectively have multiple different light emitting angles, the narrow light field excitation light elements at different positions can be controlled through the control unit to obtain the required light emission angle; the narrow light field excitation light elements can also be set on the base, and the cover can also be used The arc curve of the body makes the beams emitted by the narrow light field excitation light elements at different positions can be reflected by the cover to different angles. When it is installed on the base, an additional layer of heat conduction layer can also be set on the base. The heat generated by the narrow light field excitation light element can be conducted to the outside by the heat dissipation layer of the cover body or the heat conduction layer of the base, so that the temperature in the accommodating space is not easy to rise, and the interference when observing living organisms is reduced, and the light can be adjusted The structure is simple, which can not only reduce the production cost, but also increase the service life of the product.

On the other hand, the fluorescence observation device has the characteristic of simplifying the operation, and can be easily connected to the known optical microscope or micro-camera and other micro-optical capture devices, which increases the application range of the micro-optic capture device, meets the expectations of consumers, and improves The efficiency and accuracy of the experiment are improved, so as to obtain the high-level microscopic information of the fluorescent object to be observed, and achieve all the above-mentioned purposes.

【Description of drawings】

Fig. 1 is the side view of the first preferred embodiment of the fluorescence micro-optical device of the present invention;

Fig. 2 is a partially enlarged side view of the fluorescence micro-optical device of Fig. 1, illustrating the structure of the fluorescence observation device;

Fig. 3 is a block diagram of the fluorescence micro-optical device in Fig. 1, illustrating the connection between the control unit and the multi-angle light emitting unit;

Fig. 4 is a top view of the extended flexible driving plate of the fluorescence observation device in Fig. 2;

Fig. 5 is a side view of a group of water-cooled tubes embedded in the heat dissipation layer of the fluorescent micro-optical device in Fig. 1;

Fig. 6 is a side view of a second preferred embodiment of the fluoroscopic observation device, which can be placed on a desktop and used alone;

Fig. 7 is a side view of a preferred embodiment of a pedestal for a fluorescence micro-optical device, illustrating that the base and the cover are combined with each other, and the narrow light field excitation light element is arranged on the base;

FIG. 8 is a block diagram of the fluorescence micro-optical device of FIG. 7, illustrating the connection between the control unit and the multi-angle light emitting unit;

Fig. 9 is a top view of the circuit board arranged in the base of Fig. 7;

Figure 10 is a side view of a known optical auxiliary device; and

Fig. 11 is a side view of a known optical auxiliary mask.

【Symbol Description】

5, 5'...Fluorescent object to be observed 1...Fuselage

2...Objective lens 3...Fluorescence observation device

32, 32”…base 34, 34’, 34”…cover

36, 36”…multi-angle lighting unit 38, 38”…control unit

30...Flexible drive board 322...Loading observation area

340, 340’, 340”…accommodating space 342, 342’…observation hole

344, 344’…Light-transmitting and heat-insulating layer 346, 346”…Heating layer

362, 362’, 362”…Narrow light field excitation light element 31…Water-cooled tube

324”…thermal layer 360”…circuit board

326”…thermal insulation light transmission layer 341”…reflective surface

348’…support side edge 341’…filter

360…conducting circuit board 344”…light-transmitting reflective layer

72...ultraviolet light source 74... bellows tube

76...petri dish 78...objective lens

80...Adjustment piece 82...Mask

84...base 86...light shielding chamber

88...Light source assembly 70...Auxiliary device

6’…desktop

【Detailed ways】

The aforementioned and other technical contents, features and functions of the present invention will be clearly presented in the following detailed description of the preferred embodiments in conjunction with the accompanying drawings; the same or similar elements will be marked with similar symbols.

Fluorescence micro-optical device of the present invention, as shown in Figure 1, Figure 2 and Figure 3, is to detect a fluorescence to-be-observed object 5, mainly comprises fuselage 1, objective lens 2 and fluorescence observation device 3, and wherein objective lens 2 is to install It is arranged on the body 1, and the fluorescence observation device 3 includes a base 32, a cover body 34, a multi-angle light emitting unit 36, a control unit 38 and a flexible driving board 30, and the base 32 has a carrying observation area 322 , and be covered by the cover body 34, so that the cover body 34 and the base 32 jointly form an accommodating space 340, and on the cover body 34, an observation hole 342 is formed, which can be used for the corresponding assembly of the objective lens 2.

The cover body 34 includes a light-transmitting heat-insulating layer 344 and a heat-dissipating layer 346, and the light-transmitting heat-insulating layer 344 is arranged facing the direction of the accommodating space 340. In this example, the light-transmitting heat-insulating layer 344 is exemplified as acrylic material, and the heat-dissipating layer 346 is towards the direction away from the accommodating space 340, and the side of the inner side of the heat dissipation layer 346 is completely covered with the light-transmitting heat-insulating layer 344, and the material of the heat-dissipating layer 346 is selected to have a higher thermal conductivity than the light-transmitting heat-insulating layer 344 material, and at least in the range of visible light, its light transmittance is lower than that of the light-transmitting and heat-insulating layer 344. In this example, the heat-dissipating layer 346 is made of a metal material as an example.

The multi-angle light emitting unit 36 includes a plurality of narrow light field excitation light elements 362, and is installed in the cover body 34. The narrow light field excitation light element 362 defined here means that the light field of its light emission is limited to a predetermined The solid angle range, for example, the light emitting angle is within a solid angle of tens of degrees, instead of emitting light in all directions with a 360-degree solid angle. Also refer to FIG. 4 , the narrow light field excitation light element 362 is a plurality of LED elements in this example, and each LED element is firstly welded on a hard-type conductive circuit board 360 respectively, and then through the conductive circuit The board 360 is connected to the flexible driving board 30, and the shape of the flexible driving board 30 in this example extends in a star shape, and conducts heat to contact the surface of the heat dissipation layer 346 of the cover body 34, so that the aforementioned narrow light field excites light. The beams emitted by the element 362 are emitted toward the accommodating space 340 inside the cover body 34 , and pass through the light-transmitting and heat-insulating layers 344 respectively.

Since the sides of the cover body 34 in this example extend in an arc shape, the narrow light field excitation light element 362 installed in the cover body 34 will be affected by the structure, so that a plurality of narrow light fields arranged at different side positions The field excitation light element 362 generates different light emitting angles respectively. And each narrow light field excitation light element 362 of this example can be controlled by the control unit 38, by driving the narrow light field excitation light element 362 in different positions, adjust the irradiation angle of the narrow light field excitation light element 362, for example, this For example, the narrow light field excitation light element 362 arranged in the cover body 34 is divided into upper, middle and lower layers, and the control unit 38 controls the narrow light field excitation light element 362 excitation light source located in different curved layers, so that the narrow light field The illumination angle of the excitation light element can be controlled as required.

Since the multi-angle light-emitting unit 36 of the present invention emits light near the object to be observed, the light path of this close-to-light source is very short, and the light dissipation is less, so the high-brightness excitation beam that must be used in the past fluorescence microscope can be avoided. , and also avoid the high heat accompanying the prior art, so that the light source used in the fluorescence microscope or fluorescence camera of the present invention will not cook the object to be observed.

On the other hand, when the fluorescent object to be observed 5 is to be detected, the fluorescent object to be observed 5 is first placed on the carrying observation area 322 of the base 32, and the cover 34 is covered on the base 32, so that the fluorescent object to be observed 5 It is accommodated in the accommodating space 340, and the control unit 38 controls the narrow light field excitation light element 362 to excite the light source to stimulate the fluorescent object 5 to generate fluorescence, so that the observer can directly observe the fluorescent object 5 through the observation hole 342 through the objective lens 2. The fluorescent reaction emitted by the object 5 is observed, and the observed image of the object to be observed is captured.

As mentioned above, when the present invention adopts low-angle light emission, for example, the angle between the emitted light beam and the horizontal is less than 45 degrees, at this time, there will not be a large amount of directly reflected excitation light being incident back to the objective lens 2, and only a small amount of excitation light at most Due to the interference of noise caused by diffuse reflection, the fluorescent information, which is much weaker than the excitation light, is also easier to be observed and recorded because the signal noise is better. Compared with the existing fluorescence microscope, its optical design is to allow the light beam to directly irradiate the object to be observed along the direction of the objective lens, which cannot avoid the interference of the direct reflection of the excitation light, making the signal-to-noise ratio extremely poor, and also forcing the rear end to be installed unnecessary There are a large number of optical processing or electronic processing devices, so the present invention can make the detection data clearer, and directly match with the existing optical microscope, etc., greatly improving its flexibility of use.

Due to the heat generated by the flexible driving board 30 and the narrow light field excitation light element 362, by directly contacting the heat dissipation layer 346, it can be quickly conducted to the outer surface of the cover body 34, and diffused into the outside air, and as the present invention In the example, acrylic is used as the light-transmitting heat-insulating layer 344 , because the thermal conductivity of acrylic is much lower than that of the heat-dissipating layer 346 made of metal, which successfully prevents the temperature rise in the accommodating space 340 . In this way, adverse reactions or even death of the fluorescent object to be observed 5 due to the increase of the ambient temperature are avoided. Of course, in order to increase the heat dissipation effect, as shown in FIG. 5 , a group of heat dissipation units exemplified as water cooling tubes 31 can be buried in the heat dissipation layer 346. The water cooling tubes 31 are at least partially buried in the heat dissipation layer 346, and by continuously providing water flow , so that the heat energy transmitted from the heat dissipation layer 346 is surely led out by the water in the water cooling pipe 31 , and the heat dissipation effect of the heat dissipation layer 346 is greatly improved.

When the object to be observed is a larger animal such as a fluorescent mouse, the fluorescence observation device of the present invention can also be implemented in another embodiment, as shown in Figure 6. At this time, a larger animal such as a mouse will be used The cover body 34' of the cage is used alone, and the fluorescent object to be observed 5' is directly placed on a desktop 6', and the cover body 34' is covered on the desktop 6' to form an accommodation space 340', and the The fluorescent object to be observed 5' is accommodated in it, and then the inspector drives the narrow light field excitation light element 362' to emit light, and observes directly from the observation hole 342' whether the fluorescent object to be observed 5' has a fluorescent reaction, and when the fluorescent object to be observed 5' When the fluorescent mouse is exemplified, since the fluorescent mouse can move freely in the accommodation space 340', the light-transmitting and heat-insulating layer 344' of the cover body 34' can provide heat insulation protection, preventing the fluorescent mouse from directly touching the narrow light field for excitation The light element 362' is burnt, and it can also prevent the fluorescent mouse from directly biting the light source or polluting the equipment with excrement.

On the other hand, the observation hole 342' of this example faces the side edge of the accommodating space 340', and a supporting side edge 348' protrudes further, and another filter 341' is set at the observation hole 342' and is supported. The support of the side edge 348' and the setting of the filter 341' are to block the penetration of non-predetermined fluorescent wavelength light such as excitation light, so as to facilitate the detection personnel to observe the fluorescent reaction.

Of course, the fluorescence observation device of the present invention can be directly installed and built into a group of microscope bases, and the user can install an existing general optical microscope body on the base to match a fluorescence microscope that can observe fluorescence. microscope. The preferred embodiment of the pedestal of the present invention, as shown in Figure 7 and Figure 8, in this example, the pedestal is made up of a base 32" and a cover 34", wherein the base 32" includes a thermal insulation transparent The optical layer 326" and the heat-conducting layer 324", wherein the heat-insulating light-transmitting layer 326" can also be made of acrylic material, and the heat-conducting layer 324" is set on the side of the heat-insulating light-transmitting layer 326" and away from the accommodating space 340". direction, and the thermal conduction layer 324" can also be made of metal material, so that the thermal conductivity of the thermal conduction layer 324" is higher than that of the heat-insulating light-transmitting layer 326".

The multi-angle light-emitting unit 36" of this example further includes a circuit board 360", which is arranged on the heat-conducting layer 324" in a heat-conducting contact mode, and is provided for conducting and connecting the narrow light-field excitation light element 362", so that the narrow light-field excitation The light beams emitted by the light element 362" pass through the heat-insulating light-transmitting layer 326" and excite the light source toward the accommodating space 340". " leads to the outside.

Referring to FIG. 9 together, the narrow light field excitation light element 362" of this example is arranged in three different ring positions in the base 32", in the middle and outside, and is controlled by the control unit 38" to be located in different positions respectively. The narrow light field at the position excites the light element 362", and the side of the heat dissipation layer 346" of the cover body 34" facing the light-transmitting reflective layer 344" is polished to form a smooth mirror surface as the reflective surface 341", when the narrow light When the light beam emitted by the field excitation element 362" hits the reflective surface 341", since the side of the cover body 34" presents an arc-shaped extension, the reflection angles of different incident light beams are different from each other. Therefore, the light beam can pass through the control unit 38 "Control the narrow light field excitation light element 362" located at different ring positions to excite the light source to obtain the desired light source. Especially in this example, even the LEDs on the same radius can have different color lights, for example, The LEDs in the circle are alternately arranged with ultraviolet light and green light respectively, so that there are two kinds of fluorescent genes to be observed at the same time during the experiment.

In this example, the six LEDs in the innermost circle are white LEDs to provide normal visible light illumination, so that the operator can clearly observe, for example, how many zebrafish are there in the petri dish. Of course, as those skilled in the art can It is easy to understand that the number or arrangement position of the narrow light field excitation light elements described in each embodiment of the present invention is for the convenience of description, and does not limit the implementation of the present invention.

In summary, the fluorescence observation device, the pedestal for the fluorescence micro-optical device and the device of the present invention can control the excitation light source of the narrow light field excitation light element at different positions through the control unit, so that the narrow light field arranged on the cover body The light field excitation light element emits light sources at different angles, or the narrow light field excitation light element is placed on the base, and the light source is reflected to different angles through the cover, which simplifies the light adjustment structure, not only reduces production costs, At the same time, it can also increase the service life of the product.

Moreover, the heat generated by the narrow light field excitation light element, the flexible driving board and the circuit board can be conducted away to the outside through the heat dissipation layer of the cover or the heat conduction layer of the base, so that the temperature in the accommodating space can be controlled. It is stably controlled so that living organisms are not easily disturbed by temperature changes, increasing the efficiency and accuracy of the experiment. When the fluorescent object to be observed is a fluorescent mouse, the light-transmitting thermal insulation layer can be used as heat insulation protection to prevent the fluorescent mouse from being directly affected. Touching the narrow light field to excite the light element will cause burns, which will affect the physiological response of the fluorescent mouse or pollute and damage the light source.

Most importantly, since the fluorescence observation device provides a near-field light source, the loss of the excitation light during transmission is reduced, and because the excitation light is operated in a dark-field illumination mode, a large amount of directly reflected excitation light noise is further eliminated, which not only simplifies the design of the light source , greatly improve the signal-to-noise ratio, so that the fluorescence observation device of the present invention can be used alone, and can also be easily connected to micro-optical pickup devices such as known optical microscopes or micro-cameras, increasing the efficiency of the micro-optic pickup device. The scope of application meets the expectations of consumers and achieves all the above-mentioned purposes.

But the above-mentioned ones are only preferred embodiments of the present invention, and cannot limit the scope of the present invention with this. All simple equivalent changes and modifications made according to the claims of the present invention and the contents of the description should still be It belongs to the scope covered by the patent of the present invention.

Claims (8)

1. A fluorescence observation device with multiple groups of different-angle light sources, which is used to cover a load-bearing observation area for observing a fluorescent object to be observed located in the load-bearing observation area, wherein the fluorescence observation device includes: A cover body, together with the above-mentioned load-bearing observation area, surrounds an accommodating space for accommodating the fluorescent object to be observed, and the cover body is formed with an observation hole, and the cover body includes: A layer of light-transmitting and heat-insulating layer is arranged facing the direction of the accommodating space; and A heat dissipation layer, the thermal conductivity of which is higher than that of the above-mentioned light-transmitting heat-insulating layer, and the light transmittance at least in the range of visible light is lower than that of the above-mentioned light-transmitting heat-insulating layer. The direction of the containing space; A group of multi-angle light-emitting units, with a plurality of narrow light field excitation light elements, the light beam emitted by each of the aforementioned narrow light field excitation light elements respectively travels through the above-mentioned transparent and heat-insulating layer, and irradiates to the accommodating space, and the aforementioned narrow light field excitation the light element has at least a plurality of mutually different illumination angles; and A control unit is used to control the illumination angle of at least the narrow light field exciting light element of the multi-angle light emitting unit. 2. The fluorescence observation device as claimed in claim 1, further comprising a base, said base having said carrying observation area. 3. The fluorescence observation device as claimed in claim 2, wherein the multi-angle light-emitting unit is arranged on the base, and the side of the heat dissipation layer facing the light-transmitting reflective layer is a reflective surface for the above-mentioned multi-angle light emission The excitation light emitted by the unit is reflected and irradiated to the accommodating space; wherein the base includes: a layer of thermal insulation and light-transmitting layer, which is arranged facing the direction of the accommodating space; and a layer of heat conduction layer, whose thermal conductivity is higher than the above-mentioned The heat-insulating light-transmitting layer is arranged on one side in a direction away from the accommodating space of the heat-insulating light-transmitting layer; and the above-mentioned multi-angle light emitting unit further includes at least one circuit board, and the above-mentioned circuit board is arranged in heat-conducting contact with the heat-conducting layer. 4. The fluorescence observation device according to claim 1, further comprising at least one flexible driving board, and the aforementioned flexible driving board is thermally conductive and in contact with the above-mentioned heat dissipation layer of the cover, so that the above-mentioned narrow light-field light-emitting element is arranged on the aforementioned flexible driving board. driver board. 5. The fluorescence observation device as claimed in claim 4, further comprising a heat dissipation unit at least partially buried in the heat dissipation layer. 6. A pedestal for a fluorescent micro-optical device, which is used to observe a fluorescent object to be observed, the pedestal comprising: A base has a load-bearing observation area for observing the fluorescent object to be observed; A cover body, together with the above-mentioned base, forms an accommodating space for accommodating the fluorescent object to be observed, and the cover body is formed with an observation hole, and the cover body includes: A layer of light-transmitting and heat-insulating layer is arranged facing the direction of the accommodating space; and A heat dissipation layer, the thermal conductivity of which is higher than that of the above-mentioned light-transmitting heat-insulating layer, and the light transmittance at least in the range of visible light is lower than that of the above-mentioned light-transmitting heat-insulating layer. The direction of the containing space; A group of multi-angle light-emitting units, with a plurality of narrow light field excitation light elements, the light beam emitted by each of the aforementioned narrow light field excitation light elements respectively travels through the above-mentioned light-transmitting and heat-insulating layer, and irradiates to the accommodating space, and the aforementioned narrow light field excitation the light element has at least a plurality of mutually different illumination angles; and A control unit is used to control the illumination angle of at least the narrow light field exciting light element of the multi-angle light emitting unit. 7. The pedestal according to claim 6, further comprising at least one flexible driving board and a heat dissipation unit, and the flexible driving board is in contact with the above-mentioned heat dissipation layer of the cover for heat conduction, and is provided for the above-mentioned narrow light-field light-emitting element On the aforementioned flexible driving board, the heat dissipation unit is a water-cooled pipe at least partially buried in the heat dissipation layer. 8. A fluorescent micro-optical device for observing a fluorescent object to be observed, comprising: a fuselage; an objective lens mounted on the body; and A fluorescence observation device, comprising: A base has a load-bearing observation area for observing the fluorescent object to be observed; A cover body, together with the above-mentioned base, forms an accommodating space for accommodating the fluorescent object to be observed, and the cover body is formed with an observation hole, for the objective lens to pick up the fluorescent light located in the accommodating space through the observation hole. The image of the object to be observed, the cover includes: A layer of light-transmitting and heat-insulating layer is arranged facing the direction of the accommodating space; and A heat dissipation layer, the thermal conductivity of which is higher than that of the above-mentioned light-transmitting heat-insulating layer, and the light transmittance at least in the range of visible light is lower than that of the above-mentioned light-transmitting heat-insulating layer. The direction of the containing space; A group of multi-angle light-emitting units, with a plurality of narrow light field excitation light elements, the light beam emitted by each of the aforementioned narrow light field excitation light elements respectively travels through the above-mentioned transparent and heat-insulating layer, and irradiates to the accommodating space, and the aforementioned narrow light field excitation the light element has at least a plurality of mutually different illumination angles; and A control unit is used to control the illumination angle of at least the above-mentioned narrow light field exciting light element of the multi-angle light emitting unit.