CN116577924A - Focus detection coupling structure, automatic focus detection device and microscopic imaging system - Google Patents

Abstract

The application relates to a focusing coupling structure, an automatic focusing device and a microscopic imaging system, wherein a signal transmitting component and a signal receiving component are positioned on the same side of an objective lens; the emission beam of the signal emission component irradiates the sample through the 1/2 clear aperture of the objective lens; the receiving assembly receives the reflected beam of the sample through another 1/2 clear aperture of the objective lens. The sample is used for replacing the reflecting component, so that the problems of complex debugging process, high requirement on the debugging process, long debugging time, high difficulty and the like caused by the traditional mode of arranging the reflecting component are solved radically; the clear aperture of the objective lens is fully utilized and used as a passing path for signal transmission and reception, so that a transmitting light path and a receiving light path are simplified, the device is simple in structure, beneficial to miniaturization and integration, beneficial to improving anti-vibration interference capability, avoiding focus detection signal deviation caused by offset, improving focus detection efficiency and success rate, and high in stability, reliability and durability.

Description

Focus detection coupling structure, automatic focus detection device and microscopic imaging system

Technical Field

The application relates to the field of microscopic imaging, in particular to a microscopic imaging coupling structure, an automatic microscopic imaging device and a microscopic imaging system.

Background

In microscopic imaging application, the observed sample structure is tiny, a high-magnification objective lens is generally needed, and because of high resolution and small depth of field, focal planes of different sample areas are inconsistent, so that the condition of image acquisition blurring occurs, and the detection speed and the image acquisition quality are affected. Therefore, an automatic focusing device is needed to assist in application, and high-precision real-time dynamic focusing is realized in the testing process. The detector is utilized to obtain the signal difference reflected by the sample at the focal plane and different defocusing positions, the defocusing amount is calculated, and the focus detection is realized, so that the method is a mature automatic focus detection technology at present, and the simple example is described as follows.

Chinese patent publication No. CN109283672a discloses an automatic focusing device comprising: an emission module including a light source; a reflection module comprising a mirror, the edge of which is tangential to the optical axis of the light source for generating a semicircular eccentric light beam; the collimating mirror module comprises a collimating mirror, the collimating mirror is arranged between the reflecting mirror and the optical path of the objective lens, the central axis of the collimating mirror coincides with the optical axis of the objective lens, and the light source coincides with the reflecting focus of the collimating mirror; and a receiving module including a photosensor, a signal processing circuit, and a signal output terminal. According to the application, the edge of the reflecting mirror is tangential to the optical axis of the light source, so that the light source and the photoelectric sensor can be positioned at the focus of the collimating mirror, and in addition, the defocus amount is calculated by processing and calculating the reflected light of the measuring light, so that the position of the objective lens is quickly adjusted, the time required for focusing is reduced, and the automatic focusing speed and accuracy are improved.

Chinese patent publication No. CN207675646U discloses a focusing module for a gene sequencer, comprising an incident light path and a reflection light path; the incident light path comprises an LED point light source, a mask positioned in front of the LED point light source, a narrow-band filter, a collimating lens, a knife edge reflector and an imaging objective lens, wherein the narrow-band filter, the collimating lens, the knife edge reflector and the imaging objective lens are positioned on the incident light path; the reflection light path comprises an imaging objective lens, a knife-edge reflector, a collimating lens, a narrow-band filter and a two-dimensional PSD sensor, wherein the collimating lens and the narrow-band filter are positioned on the reflection light path, light spots of a specific pattern irradiate onto fluorophores of the biochip, the obtained reflection light spots pass through the imaging objective lens to form parallel light beams, and the part of the parallel light beams, which passes through the knife-edge of the knife-edge reflector, sequentially passes through the collimating lens and the narrow-band filter and then reaches the two-dimensional PSD sensor; the two-dimensional PSD sensor is connected with a signal processing circuit, and the signal processing circuit obtains characteristic information of the imaging light spots.

The technical solutions of the two patent documents each include a signal transmitting assembly, a reflecting assembly and a receiving assembly, wherein the reflecting assembly mainly uses a reflecting mirror, such as a right-angle reflecting prism or a plane reflecting mirror, to reflect the transmitted light beam, change the transmission direction thereof and form a semicircular eccentric light beam. The transmitting component, the reflecting component and the receiving component are arranged in a T shape or inverted T shape, namely inverted T shape, so that the structural volume is increased, the device has more components and large structure, and therefore, the miniaturization design cannot be carried out, and the miniaturization integration is not facilitated; the reflection assembly has extremely high sensitivity, the debugging process is complex, the requirement on the debugging process is high, the debugging time is long, and the difficulty is high; in addition, in practical application, the anti-vibration interference capability is lower, and deviation is easy to generate, so that focus detection signal deviation is easy to generate, and focus detection failure is caused, and the stability, reliability and durability are lower.

Disclosure of Invention

Based on this, it is necessary to provide a focus detection coupling structure, an automatic focus detection device and a microscopic imaging system.

In one embodiment, a focus detection coupling structure includes a signal transmitting component and a receiving component, and further includes a transmitting light path and a receiving light path:

the signal transmitting component and the receiving component are positioned on the same side of the objective lens;

the emission light path includes: the emission light beam of the signal emission component irradiates the sample through the 1/2 clear aperture of the objective lens;

the receiving optical path includes: the receiving assembly receives the reflected beam of the sample through another 1/2 clear aperture of the objective lens.

According to the focus detection coupling structure, on one hand, the sample is used for replacing the reflecting component, so that the debugging process is simple, and the problems of complex debugging process, high requirement on the debugging process, long debugging time, high difficulty and the like caused by the fact that the reflecting component is arranged in a traditional mode are solved from the root; on the other hand, the clear aperture of the objective lens is fully utilized and is used as a passing path for signal transmission and reception, so that a transmitting light path and a receiving light path are simplified, the device is simple in structure, beneficial to miniaturization and integration, beneficial to improving the anti-vibration interference capability, and capable of avoiding focus detection signal deviation caused by offset, thereby improving the focus detection efficiency and success rate, and high in stability, reliability and durability.

Further, in one embodiment, the sample is used as a reflecting component to form the receiving light path.

In one embodiment, the signal emitting assembly includes a light source and a collimating element;

the light source is used for outputting the emission light beam, and the emission light beam is processed by the collimating element to form a parallel light beam to irradiate the objective lens.

In one embodiment, the signal emitting assembly further comprises a diaphragm disposed between the light source and the collimating element; and/or the number of the groups of groups,

the signal transmitting assembly further comprises an optical filter, wherein the optical filter is arranged between the collimating element and the objective lens or between the light source and the collimating element.

In one embodiment, the receiving assembly includes a first focusing element and a first detector;

the first focusing element is used for receiving the reflected light beam and converging the reflected light beam to the first detector;

the first detector is used for detecting and outputting the optical signal of the reflected light beam.

In one embodiment, the receiving assembly further comprises a diaphragm disposed between the first focusing element and the first detector; and/or the number of the groups of groups,

the receiving assembly further comprises an optical filter arranged between the first focusing element and the objective lens or between the first focusing element and the first detector.

In one embodiment, an automatic focus detection device comprises an industrial personal computer and the focus detection coupling structure of any embodiment;

the industrial personal computer is respectively connected with the signal transmitting assembly and the receiving assembly and is used for controlling the signal transmitting assembly to output a transmitting light beam, processing a reflected light beam of the receiving assembly and calculating according to the reflected light beam to obtain a focal plane offset;

the industrial personal computer is also used for connecting with an objective lens, and the objective lens is controlled to move according to the focal plane offset to perform focus detection.

According to the automatic focus detection device, on one hand, the sample is used for replacing the reflection assembly, so that the debugging process is simple, and the problems of complex debugging process, high requirement on the debugging process, long debugging time, high debugging difficulty and the like are avoided; on the other hand has simplified emission light path and receiving light path, not only simple structure is favorable to miniaturized integration, is favorable to promoting anti-vibration interference ability moreover, avoids leading to examining burnt signal deviation because of the skew to automatic burnt efficiency and success rate have been examined in the promotion, and have higher stability, reliability and durability.

In one embodiment, the automatic focusing device further comprises an objective lens table connected with the industrial personal computer, wherein the objective lens table is used for fixedly mounting the objective lens, and the industrial personal computer is used for changing the distance between the objective lens and the sample by controlling the movement of the objective lens table and is used for focusing the sample; and/or the number of the groups of groups,

the automatic focus detection device further comprises a moving device connected with the industrial personal computer, the moving device is used for fixedly installing a sample, and the industrial personal computer is used for adjusting the position of the sample by controlling the moving device to move.

In one embodiment, the automatic focusing device further comprises a reference component connected with the industrial personal computer, wherein the reference component comprises a spectroscope, a second focusing element and a second detector;

the emission light beam of the signal emission component passes through the spectroscope, part of the emission light beam irradiates the sample through the 1/2 clear aperture of the objective lens, and the rest part irradiates the second detector through the second focusing element;

the industrial personal computer also processes the optical signal of the second detector to obtain a reference signal so as to reduce fluctuation interference of the optical power emitted by the light source.

In one embodiment, a microscopic imaging system includes an objective lens and an automated focusing apparatus of any of the embodiments.

In one embodiment, the microscopy imaging system is a gene sequencer.

Drawings

In order to more clearly illustrate the technical solutions of the embodiments or the conventional techniques of the present application, the drawings required for the descriptions of the embodiments or the conventional techniques will be briefly described below, and it is apparent that the drawings in the following descriptions are only some embodiments of the present application, and other drawings may be obtained according to the drawings without inventive effort for those skilled in the art.

Fig. 1 is a schematic diagram illustrating an application of an embodiment of a focus detection coupling structure according to the present application.

Fig. 2 is a schematic diagram of an emission light path of the embodiment shown in fig. 1.

Fig. 3 is a schematic diagram of a receiving optical path of the embodiment shown in fig. 1.

Fig. 4 is a schematic application diagram of another embodiment of the focus detection coupling structure according to the present application.

FIG. 5 is a schematic diagram showing the position change of the light spot on the detector at different focal plane offsets according to the embodiment shown in FIG. 4.

FIG. 6 is a graph showing the relationship between the focal plane offset and the signal difference.

Fig. 7 is a schematic diagram illustrating an application of another embodiment of the focus detection coupling structure according to the present application.

Fig. 8 is a schematic diagram illustrating an application of an embodiment of an automatic focusing apparatus according to the present application.

Fig. 9 is a schematic diagram illustrating an application of another embodiment of the automatic focusing apparatus of the present application.

Fig. 10 is a schematic view illustrating an application of another embodiment of the automatic focusing apparatus of the present application.

Reference numerals:

a focus detection coupling structure 100;

a signal emitting assembly 10, a light source 101, a collimating element 102;

a receiving assembly 20;

an emission beam 30, a reflection beam 40;

a reference assembly 50, a beam splitter 501, a second focusing element 502, a second detector 503;

objective lens 200, first focusing element 201, first detector 202, spot 203, first pixel 204, second pixel 205;

sample 300, mobile device 301;

the industrial personal computer 400.

Detailed Description

In order that the above objects, features and advantages of the application will be readily understood, a more particular description of the application will be rendered by reference to the appended drawings. In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present application. The present application may be embodied in many other forms than described herein and similarly modified by those skilled in the art without departing from the spirit of the application, whereby the application is not limited to the specific embodiments disclosed below.

It will be understood that when an element is referred to as being "mounted" or "disposed" on another element, it can be directly on the other element or intervening elements may also be present. When a component is considered to be "connected" to another component, it can be directly connected to the other component or intervening components may also be present. The terms "vertical", "horizontal", "upper", "lower", "left", "right" and the like are used in the description of the present application for the purpose of illustration only and do not represent the only embodiment.

Furthermore, the terms "first," "second," and the like, are used for descriptive purposes only and are not to be construed as indicating or implying a relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defining "a first" or "a second" may explicitly or implicitly include at least one such feature. In the description of the present application, the meaning of "plurality" means at least two, for example, two, three, etc., unless specifically defined otherwise.

In the present application, unless expressly stated or limited otherwise, a first feature "up" or "down" on a second feature may be that the first feature is in direct contact with the second feature, or that the first feature and the second feature are in indirect contact through intermedial media. Moreover, a first feature "above," "over" and "on" a second feature may be a first feature directly above or obliquely above the second feature, or simply indicate that the first feature is higher in level than the second feature. The first feature being "under", "below" and "beneath" the second feature may be the first feature being directly under or obliquely under the second feature, or simply indicating that the first feature is less level than the second feature.

Unless defined otherwise, all technical and scientific terms used in the specification of the present application have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used in the description of the application herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the application. The term "and/or" as used in the description of the present application includes any and all combinations of one or more of the associated listed items.

The application discloses a focus detection coupling structure, an automatic focus detection device and a microscopic imaging system, which comprise part of structures or all structures of the following embodiments; that is, the focus detection coupling structure, the automatic focus detection device, and the microscopic imaging system include some or all of the following technical features. In one embodiment of the application, a focus detection coupling structure comprises a signal transmitting component, a receiving component, a transmitting light path and a receiving light path: the signal transmitting component and the receiving component are positioned on the same side of the objective lens; the emission light path includes: the emission beam 30 of the signal emission component irradiates the sample through the 1/2 clear aperture of the objective lens; the receiving optical path includes: the receiving assembly receives the reflected beam 40 of the sample through another 1/2 clear aperture of the objective lens. According to the focus detection coupling structure, on one hand, the sample is used for replacing the reflecting component, so that the debugging process is simple, and the problems of complex debugging process, high requirement on the debugging process, long debugging time, high difficulty and the like caused by the fact that the reflecting component is arranged in a traditional mode are solved from the root; on the other hand, the clear aperture of the objective lens is fully utilized and is used as a passing path for signal transmission and reception, so that a transmitting light path and a receiving light path are simplified, the device is simple in structure, beneficial to miniaturization and integration, beneficial to improving the anti-vibration interference capability, and capable of avoiding focus detection signal deviation caused by offset, thereby improving the focus detection efficiency and success rate, and high in stability, reliability and durability.

Further, in each embodiment, the focus detecting coupling structure and the automatic focus detecting device with the focus detecting coupling structure can be applied to a microscopic imaging system such as a gene sequencer, for example, in microscopic imaging application, an observed sample structure is tiny, a high-magnification objective lens is generally needed, and due to high resolution and small depth of field, focal planes of different sample areas are inconsistent, so that the condition of image acquisition blurring occurs, and the detection speed and the image acquisition quality are affected. Therefore, an automatic focusing device is needed to assist in application, and high-precision real-time dynamic focusing is realized in the testing process. However, as described above, the structural design of the reflective component of the conventional automatic focusing device causes problems such as difficult debugging, complicated structure and difficult focusing.

In one embodiment, as shown in fig. 1, an application of a focusing coupling structure 100 is shown, where the focusing coupling structure 100 includes a signal transmitting component 10 and a receiving component 20, where the signal transmitting component 10 and the receiving component 20 are located on the same side of an objective lens 200; the focus detection coupling structure 100 further includes a transmitting light path and a receiving light path; the focusing coupling structure 100 can emit a signal to irradiate the sample 300 through the objective lens 200 and receive the signal reflected by the sample 300. As shown in fig. 1, the signal transmitting element 10 and the receiving element 20 are located on the same side of the objective lens 200, i.e. the signal transmitting element 10 and the receiving element 20 are located on one side of the objective lens 200 at the same time, and the sample 300 to be used is located on the other side of the objective lens 200. The structure design is innovative to couple the transmitting component and the receiving component, omits the reflecting component relative to the traditional technology, takes the clear aperture of the objective lens as a passing path for signal transmission and reception, completely solves the defects caused by the existence of the reflecting component, and has the advantages of simple structure, contribution to miniaturization and integration, simple debugging process, high stability and reliability and the like.

In various embodiments, the emission light path is as shown in fig. 2, and the emission light path includes: the emission beam 30 of the signal emission assembly 10 is irradiated onto the sample 300 through the 1/2 clear aperture of the objective lens 200; clear Aperture (Clear Aperture) is also known as the effective Aperture, and is used to represent the size of the Clear Aperture. Further, in each embodiment, the clear aperture of the objective lens 200 is divided into two parts in application, i.e. the clear aperture is essentially a whole, and is not physically changed, but is artificially defined in application to be divided into two parts which are substantially equivalent, one part passing the emitted light beam 30 as a 1/2 clear aperture and the other part passing the reflected light beam 40 as another 1/2 clear aperture, i.e. passing the reflected light beam 40 in opposite directions with respect to the emitted light beam 30.

In various embodiments, the receiving optical path is as shown in fig. 3, and the receiving optical path includes: the receiving assembly 20 receives the reflected beam 40 of the sample 300 through another 1/2 clear aperture of the objective lens 200. Further, in one embodiment, the sample 300 is used as a reflective component to form the receiving optical path. According to the structural design, the sample is used for replacing the reflecting component to form the reflecting light beam 40, the receiving light beam 40 is used for forming the receiving light path, the problems of complex debugging process, high debugging process requirement, long debugging time, high difficulty and the like caused by the fact that the reflecting component is arranged in a traditional mode are fundamentally solved, and compared with a traditional focusing device, the debugging process is simple, quick and easy to realize.

As shown in fig. 4, a focus detection coupling structure 100, in which a signal transmitting assembly 10 is composed of a light source 101 and a collimating element 102, and a receiving assembly 20 is composed of a first focusing element 201 and a first detector 202. The optical signal emitted from the light source 101 is irradiated onto the objective lens 200 in a parallel beam through the collimator element 102, and is focused on the sample 300 through the objective lens 200. Wherein the sample 300 is loaded on a mobile device 301. The optical signal reflected by the sample 300 is parallel to the first focusing element 201 after passing through the objective lens 200, and is irradiated onto the photosensitive surface of the first detector 202 through the first focusing element 201. The first detector 202 processes and outputs the obtained optical signal, and in combination with fig. 8, the focal plane offset is obtained after processing, analyzing and calculating by the industrial personal computer 400, and the objective lens is controlled to move to realize the function of focus detection.

In this embodiment, the signal transmitting assembly 10 and the receiving assembly 20 are spatially located on the same side. The light beam emitted from the signal emitting assembly 10 irradiates through the 1/2 clear aperture of the objective lens 200 and onto the sample 300; the beam reflected from the sample 300 passes through the other 1/2 clear aperture of the objective lens 200 and is detected by the receiving assembly 20. Further, in one embodiment, the optotype coupling structure does not include a reflection component, an objective lens 200 and a sample 300, but includes only the signal transmitting component 10 and the receiving component 20, and the transmitting light path and the receiving light path formed by the signal transmitting component 10 and the receiving component 20 in cooperation with the objective lens 200 and the sample 300. The structure design fully utilizes the clear aperture of the objective lens, takes the objective lens as a passing path for signal transmission and receiving, and is matched with the design of replacing a reflecting component by utilizing a sample, so that the transmitting light path and the receiving light path are greatly simplified, the structure has the advantage of simple structure, miniature structural parts which are easy to realize miniaturized integration are greatly improved, the application field of products is greatly improved, and the large-scale integrated design scheme obtained by adopting the focus detection coupling structure is beneficial to improving the vibration interference resistance, avoiding focus detection signal deviation caused by offset and has higher stability, reliability and durability.

In one embodiment, as shown in fig. 4, the signal emitting assembly 10 includes a light source 101 and a collimating element 102; the light source 101 is configured to output the emitted light beam 30, and the emitted light beam is processed by the collimating element 102 to form a parallel light beam to irradiate the objective lens 200. Further, in one embodiment, the light source 101 in the emitting component may be a xenon lamp, a halogen lamp, an LED, a laser or other light source, and the type is not particularly limited; in practical applications, the wavelength of the light source 101 needs to avoid overlapping with the wavelength of the excitation light and the fluorescence emitted from the sample 300. Further, in one embodiment, the collimating element 102 may be a single-piece or multi-piece lens, and is not particularly limited, depending on the specific requirements and application scenario.

In one embodiment, the signal emitting assembly 10 further comprises a diaphragm disposed between the light source 101 and the collimating element 102. In one embodiment, the signal emitting assembly 10 further includes a filter disposed between the collimating element 102 and the objective lens 200, or between the light source 101 and the collimating element 102. In one embodiment, the signal emitting assembly 10 further includes a light filter and a diaphragm disposed between the light source 101 and the collimating element 102, wherein the light filter is disposed between the collimating element 102 and the objective lens 200, or between the light source 101 and the collimating element 102. The rest of the embodiments are analogized and will not be described in detail. Specifically, in one embodiment, the signal emitting component 10 formed by the light source 101 and the collimating element 102 may have a diaphragm added to the front end of the light source 101 according to actual needs, and the diaphragm light-passing hole may be circular, square or other shapes; meanwhile, a filter or the like may be added before or after the collimating element 102 for processing the emitted light beam 30 of the light source 101.

In one embodiment, as shown in fig. 4, the receiving assembly 20 includes a first focusing element 201 and a first detector 202; the first focusing element 201 is configured to receive the reflected light beam 40 and converge to the first detector 202; the first detector 202 is configured to detect and output the optical signal of the reflected light beam 40. Further, in one embodiment, the first focusing element 201 may be a single-piece or multi-piece lens, and is not limited in particular, depending on the specific requirements and application. Further, in one embodiment, the first detector 202 is a (photo diode, PD) detector, which may be a single-pixel, two-pixel or multi-pixel detector, and is not limited in particular.

In one embodiment, the receiving assembly 20 further comprises a diaphragm disposed between the first focusing element 201 and the first detector 202; in one embodiment, the receiving assembly 20 further includes a filter disposed between the first focusing element 201 and the objective lens 200 or between the first focusing element 201 and the first detector 202. Specifically, in one embodiment, the receiving component 20 formed by the first focusing element 201 and the first detector 202 may have a diaphragm added to the front end of the first detector 202 according to actual needs, and the diaphragm light-passing hole may be circular, square or other shapes; meanwhile, a filter may be added before or after the first focusing element 201 for processing the light signal reflected from the sample 300.

Specifically, in one embodiment, as shown in fig. 5, the first detector 202 is a two-pixel photodiode, the first pixel 204 detects the first signal I1, and the second pixel 205 detects the second signal I2. When the distance between the sample 300 and the objective lens 200 changes, that is, the focal plane position of the sample 300 at the objective lens 200 or the focal plane position is deviated, the light beam reflected from the sample 300 irradiates on different positions of the photosensitive surface of the first detector 202, and the detected first signal I1 and the second signal I2 change. Fig. 5 shows the change in position of the spot 203 on the first detector 202 as the sample 300 moves from a position away from the focal plane of the objective lens to a position closer to the focal plane of the objective lens. Thus, the relationship between the focal plane offset (focal plane offset, FPO) and the signal difference (Intensity difference, i_diff) can be obtained as shown in fig. 6, where i_diff=i1-I2. Based on the relationship curve of the focal plane offset and the signal difference, the focal plane offset calculation can be performed by matching with the signal detected by the first detector 202 through the industrial personal computer 400 in the test process, so as to realize real-time focal detection.

In one embodiment, as shown in fig. 7, the autofocus coupling structure 100 further includes a reference component 50, and the automatic focus detecting device includes a signal transmitting component 10, a receiving component 20, and a reference component 50. The optical signal emitted by the signal emitting assembly 10 is divided into two parts, one part irradiates the reference assembly 50, the other part irradiates the sample 300 through the objective lens 200, and the receiving assembly 20 detects the signal reflected by the sample 300. Specifically, the optical signal emitted by the signal emitting component 10 or the light source 101 thereof irradiates the reference component 50 through the collimating element 102, a part of the parallel light beams irradiates the reference component 50 to form a reference signal, and the other part of the parallel light beams penetrate the objective 200 and are focused on the sample 300 through the objective 200, and the reference component 50 is adopted to overcome the influence of incomplete consistency of the output of the signal emitting component, reduce the fluctuation interference of the optical power emitted by the signal emitting component 10 or the light source 101 thereof, improve the calculation precision of the focal plane offset, and realize the focus detection function with higher precision.

In one embodiment, an automatic focusing device is shown in fig. 8, and includes an industrial personal computer 400 and the focusing coupling structure 100 according to any of the embodiments; in one embodiment, the automatic focus detection device includes an industrial personal computer 400 and a focus detection coupling structure 100, wherein the focus detection coupling structure 100 includes a signal transmitting component 10, a receiving component 20, a transmitting light path and a receiving light path: the signal transmitting component 10 and the receiving component 20 are positioned on the same side of the objective lens 200; the emission light path includes: the emission beam 30 of the signal emission assembly 10 is irradiated onto the sample 300 through the 1/2 clear aperture of the objective lens 200; the receiving optical path includes: the receiving assembly 20 receives the reflected beam 40 of the sample 300 through another 1/2 clear aperture of the objective lens 200. The rest of the embodiments are analogized and will not be described in detail.

As shown in fig. 8, the industrial personal computer 400 is respectively connected to the signal transmitting assembly 10 and the receiving assembly 20, and is configured to control the signal transmitting assembly 10 to output a transmitting beam 30, process a reflected beam 40 of the receiving assembly 20, and calculate and obtain a focal plane offset according to the reflected beam 40; the industrial personal computer 400 is further used for connecting with the objective lens 200, and performing focus detection by controlling the objective lens 200 to move according to the focal plane offset. In connection with the following embodiments, the industrial personal computer 400 is connected to the objective lens 200 in an indirect connection manner, for example, the industrial personal computer 400 is connected to the objective lens 200 through a middleware, and the position of the objective lens (200) is adjusted by controlling the middleware to move. It should be noted that, in the embodiments of the present application, the structural design of the focus detection coupling structure 100 is mainly that the focal length calculation mode is not improved, and the specific calculation mode can be implemented by using a conventional technology, which is omitted here. According to the automatic focus detection device disclosed by the embodiments of the application, on one hand, a sample is used for replacing a reflection assembly, so that the debugging process is simple, and the problems of complex debugging process, high requirement on the debugging process, long debugging time, high debugging difficulty and the like are avoided; on the other hand has simplified emission light path and receiving light path, not only simple structure is favorable to miniaturized integration, is favorable to promoting anti-vibration interference ability moreover, avoids leading to examining burnt signal deviation because of the skew to automatic burnt efficiency and success rate have been examined in the promotion, and have higher stability, reliability and durability.

In one embodiment, the automatic focusing device further includes an objective table connected to the industrial personal computer 400, the objective table is used for fixedly mounting the objective 200, the industrial personal computer 400 is used for controlling the objective table to move so as to change the distance between the objective 200 and the sample 300, and is used for focusing the sample 300, i.e. the objective 200 is mounted on the objective table, and the industrial personal computer 400 drives the objective table to move so as to drive the objective 200 to move, thereby adjusting the distance between the objective 200 and the sample 300, and realizing high-precision real-time dynamic focusing. With such a structural design, when in use, the objective lens 200 is only fixed on the object lens table, and the position of the object lens table can be conveniently adjusted by the industrial personal computer 400, so as to change the position of the objective lens 200. In various embodiments, the objective table may also be referred to as an objective lens moving device, and the objective lens 200 may be fixed on the objective table by clamping, negative pressure adsorption, limiting mounting, or the like.

In one embodiment, as shown in fig. 8, the automatic focusing device further includes a moving device 301 connected to the industrial personal computer 400, the moving device 301 is used for fixedly mounting the sample 300, and the industrial personal computer 400 is used for adjusting the position of the sample 300 by controlling the moving device 301 to move; that is, the sample 300 is mounted on the moving device 301, and the industrial personal computer 400 drives the moving device 301 to move, so as to drive the sample 300 to move. With such a structural design, the position of the mobile device 301 can be conveniently adjusted by the industrial personal computer 400 so as to change the position of the sample 300 only by fixing the sample 300 on the mobile device 301 when in use. In various embodiments, the moving device 301 may also be referred to as a sample stage, and the sample 300 may be fixed on the moving device 301 by clamping, negative pressure adsorption or accommodation.

Specifically, in one embodiment, the industrial personal computer 400 is physically connected to the light source 101, the moving device 301 carrying the objective lens 200, the moving device 301 carrying the sample 300, and the first detector 202, and controls the light source 101 to be turned on or off and controls the objective lens stage carrying the objective lens 200 and the moving device 301 carrying the sample 300 to move by an electric signal command. Meanwhile, the industrial personal computer 400 may convert, process, calculate and analyze the signal acquired by the first detector 202.

In one embodiment, as shown in fig. 9, the automatic focus detecting apparatus further includes a reference component 50 connected to the industrial personal computer 400. In this embodiment, the automatic focusing apparatus includes a signal transmitting component 10, a receiving component 20 and a reference component 50. The optical signal emitted from the optotype coupling structure 100 is divided into two parts, one part is irradiated to the reference assembly 50, and the other part is irradiated to the sample 300 through the objective lens 200. The receiving assembly 20 detects the signal reflected back from the sample 300. The reference component 50 directly receives the optical signal emitted by the focus detection coupling structure 100, and is not affected by other optical paths basically, so that the obtained reference signal can be used as a reference state of the emission beam 30, and thus when the reflected beam 40 is analyzed, the change of the emission beam 30 can be analyzed by adopting the reference signal or the reference state, thereby reducing the fluctuation interference of the light power emitted by the light source, improving the calculation precision of the focal plane offset, and realizing the focus detection function with higher precision.

As shown in fig. 10, the reference assembly 50 includes a beam splitter 501, a second focusing element 502, and a second detector 503; the emitted light beam 30 of the signal emitting assembly 10 passes through the beam splitter 501, part of the emitted light beam passes through the 1/2 clear aperture of the objective lens 200 to be irradiated onto the sample 300, and the rest of the emitted light beam passes through the second focusing element 502 to be irradiated onto the second detector 503; furthermore, the beam splitter (501) can split the emitted light beam according to different energy ratios, for example, the emitted light beam is split according to a ratio of 1:1 or 1:2, and the specific ratio is determined according to practical application conditions. And, the industrial personal computer 400 further processes the optical signal of the second detector 503 to obtain a reference signal, so as to reduce the fluctuation interference of the optical power emitted by the light source, and by adjusting the focal plane offset, the calculation accuracy of the focal plane offset is improved, and a higher-accuracy focal length detection function is realized. Unlike the embodiment shown in fig. 8, the reference component 50 is added in this embodiment, and the reference component 50 is composed of the beam splitter 501, the second focusing element 502 and the second detector 503, and in this embodiment, the reference component 50 may be used as a part of the focusing coupling structure 100 or may be separately disposed with respect to the focusing coupling structure 100.

Specifically, the optical signal emitted from the light source 101 irradiates the beam splitter 501 with a parallel light beam through the collimating element 102, one part of the light beam passes through the objective lens 200, is focused on the sample 300 through the objective lens 200, and the other part of the light beam is reflected by the beam splitter 501 to the second focusing element 502, and is focused on the second detector 503 through the second focusing element 502. Wherein the sample 300 is loaded on a mobile device 301. The optical signal reflected by the sample 300 is parallel to the first focusing element 201 after passing through the objective lens 200, and is irradiated onto the photosensitive surface of the first detector 202 through the first focusing element 201. The first detector 202 processes and outputs the obtained optical signal, and obtains the focal plane offset after processing, analyzing and calculating by the industrial personal computer 400, and controls the objective lens to move so as to realize the function of focus detection. By adopting the reference component 50, the influence of incomplete consistency of the output of the signal transmitting component can be overcome, the fluctuation interference of the light power emitted by the light source 101 can be reduced, the calculation precision of the focal plane offset can be improved, and the higher-precision focus detection function can be realized.

In one embodiment of a specific application, the first detector 202 is a two-pixel PD, and detects the first signal I1 and the second signal I2 respectively; the second detector 503 is a single pixel PD and can detect the signal Iref. When the distance between the sample 300 and the objective lens 200 changes, that is, the focal plane position of the sample 300 in the objective lens 200 or the focal plane position is deviated, the light beam reflected from the sample 300 irradiates on different positions of the photosensitive surface of the first detector 202, and the detected first signal I1 and second signal I2 change; in this process, the signal Iref detected by the second detector 503 is unchanged and is used as a reference signal. Therefore, by fitting a relation curve between the first signal I1 and the second signal I2 on the first detector 202 and the signal Iref on the second detector 503, the focal plane offset FPO can be obtained, and the focus detection function is realized; in addition, by adding the reference component 50 and using the obtained signal Iref as a reference signal, the interference of the fluctuation of the light power emitted by the light source 101 can be reduced better, the calculation precision of the focal plane offset is improved, and the higher-precision focusing function is realized.

In one embodiment, a microscopic imaging system includes an objective lens 200 and an automated focusing apparatus as described in any of the embodiments. In one embodiment, the microscopy imaging system is a gene sequencer. In the same way, the microscopic imaging system also has the advantages of simple debugging process, simple automatic focusing light path, quick automatic focusing, high success rate and the like, is beneficial to the large-scale integration of the automatic focusing device, and has higher stability, reliability and durability in the aspect of automatic focusing.

It should be noted that other embodiments of the present application further include a focus detection coupling structure, an automatic focus detection device, and a microscopic imaging system, which are formed by combining the technical features of the above embodiments.

The technical features of the above-described embodiments may be arbitrarily combined, and all possible combinations of the technical features in the above-described embodiments are not described for brevity of description, however, as long as there is no contradiction between the combinations of the technical features, they should be considered as the scope of the description.

The above examples illustrate only a few embodiments of the application, which are described in detail and are not to be construed as limiting the scope of the claims. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the application, which are all within the scope of the application. Accordingly, the scope of the application should be determined from the following claims.

Claims (10)

1. The utility model provides a examine burnt coupling structure, includes signal transmission subassembly (10) and receiving element (20), its characterized in that still includes transmission light path and receiving light path:

the signal transmitting component (10) and the receiving component (20) are positioned on the same side of the objective lens (200);

the emission light path includes: an emission beam (30) of the signal emission assembly (10) is irradiated onto a sample (300) through a 1/2 clear aperture of the objective lens (200);

the receiving optical path includes: the receiving assembly (20) receives a reflected light beam (40) of the sample (300) through another 1/2 clear aperture of the objective lens (200).

2. The optotype coupling structure of claim 1, wherein the signal emitting assembly (10) comprises a light source (101) and a collimating element (102);

the light source (101) is used for outputting the emission light beam (30), and the emission light beam is processed by the collimating element (102) to form a parallel light beam to irradiate the objective lens (200).

3. The optotype coupling structure of claim 2, wherein the signal emitting assembly (10) further comprises a diaphragm arranged between the light source (101) and the collimating element (102); and/or the number of the groups of groups,

the signal emitting assembly (10) further comprises a light filter arranged between the collimating element (102) and the objective lens (200) or between the light source (101) and the collimating element (102).

4. The optotype coupling structure of claim 1, wherein the receiving assembly (20) comprises a first focusing element (201) and a first detector (202);

-said first focusing element (201) for receiving said reflected light beam (40) and converging to said first detector (202);

the first detector (202) is configured to detect and output an optical signal of the reflected light beam (40).

5. The optotype coupling structure of claim 4, wherein the receiving assembly (20) further comprises a diaphragm disposed between the first focusing element (201) and the first detector (202); and/or the number of the groups of groups,

the receiving assembly (20) further comprises a filter arranged between the first focusing element (201) and the objective lens (200) or between the first focusing element (201) and the first detector (202).

6. An automatic focus detection device, characterized by comprising an industrial personal computer (400) and a focus detection coupling structure according to any one of claims 1 to 5;

the industrial personal computer (400) is respectively connected with the signal transmitting assembly (10) and the receiving assembly (20) and is used for controlling the signal transmitting assembly (10) to output a transmitting light beam (30) and processing a reflected light beam (40) of the receiving assembly (20), and calculating and obtaining a focal plane offset according to the reflected light beam (40);

the industrial personal computer (400) is also used for connecting an objective lens (200), and the objective lens (200) is controlled to move according to the focal plane offset to perform focus detection.

7. The automatic focus detection apparatus according to claim 6, further comprising an objective table connected to the industrial personal computer (400), the objective table being for fixedly mounting the objective (200), the industrial personal computer (400) being for performing focus detection on a sample (300) by controlling the objective table to move to change a distance between the objective (200) and the sample (300); and/or the number of the groups of groups,

the automatic focus detection device further comprises a moving device (301) connected with the industrial personal computer (400), the moving device (301) is used for fixedly installing the sample (300), and the industrial personal computer (400) is used for adjusting the position of the sample (300) by controlling the moving device (301) to move.

8. The automatic focus detection apparatus according to any one of claims 6 to 7, further comprising a reference assembly (50) connected to the industrial personal computer (400), the reference assembly (50) comprising a beam splitter (501), a second focusing element (502) and a second detector (503);

the emitted light beam (30) of the signal emitting assembly (10) passes through the spectroscope (501), part of the emitted light beam irradiates the sample (300) through a 1/2 clear aperture of the objective lens (200), and the rest part of the emitted light beam irradiates the second detector (503) through the second focusing element (502);

the industrial personal computer (400) also processes the optical signal of the second detector (503) to obtain a reference signal so as to reduce fluctuation interference of the optical power emitted by the light source.

9. A microscopic imaging system, characterized by comprising an objective lens (200) and an automatic focusing device according to any of claims 6 to 8.

10. The microscopy imaging system of claim 9, wherein the microscopy imaging system is a gene sequencer.