Illumination system and microscope device

Abstract

The present invention discloses an illumination system, comprising a light source assembly, a light-homogenizing shaping element arranged on the output light path of the light source assembly, and an objective lens group arranged on the output light path of the light-homogenizing shaping element; the light-homogenizing shaping element is used to homogenize and shape the light emitted by the light source assembly into a light beam with a predetermined light spot shape; the objective lens group is used to modulate the light spot size of the light beam output by the light source assembly. In the present application, a microlens array is used to reduce the energy loss of the illumination beam, improve the illumination intensity, and ensure the uniformity of the light spot illumination of the illumination beam, so that when the illumination system is applied to a microscope device, the noise of the detection image of the imaging system can be reduced, which is conducive to improving the accuracy of the detection image analysis result; the present application also provides a microscope device with the above-mentioned beneficial effects.

Description

Technical Field

[0001] The invention relates to the technical field of optical illumination, and in particular to an illumination system and a microscope device. Background Art

[0002] In the field of fluorescent molecule detection and analysis, microscope equipment is generally used for fluorescent image acquisition, and fluorescent molecule analysis is performed based on fluorescent image acquisition. The quality of the optical-mechanical system in the fluorescence microscope directly determines the quality of the image obtained by the image sensor. An excellent optical-mechanical system can greatly reduce the pressure for later image processing such as fluorescent molecule positioning, signal enhancement, and image denoising. In the field of fluorescent molecule detection, the optical-mechanical system can be roughly divided into an illumination system module and an imaging system module; the light beam output by the illumination light source in the illumination system module is modulated into a specific shape of light spot such as a circle or a rectangle through a series of optical elements and incident on the target surface, where fluorescent molecules or fluorescent groups are distributed on the target surface; the role of the imaging system module is to image the fluorescent signal emitted from the target surface on the industrial camera through a microscope objective.

[0003] In order to enable the optomechanical system to capture the best fluorescence signal, how to improve the matching degree between the illumination spot output by the illumination system in the microscope equipment and the effective field of view of the imaging system, and how to improve the illumination uniformity and illumination power density of the illumination spot are all issues that need to be focused on in fluorescent molecule detection. Summary of the invention

[0004] The purpose of the present invention is to provide an illumination system and a microscope device, which improve the spot illumination and spot illumination uniformity of the illumination beam to a certain extent, and provide high-quality illumination for detection image acquisition in the microscope device.

[0005] In order to solve the above technical problems, the present invention provides a lighting system, comprising:

[0006] A light source assembly, a light homogenizing and shaping element arranged on an output light path of the light source assembly, and an objective lens group arranged on an output light path of the light homogenizing and shaping element;

[0007] The light homogenizing and shaping element is used to homogenize and shape the light emitted by the light source assembly into a light beam with a predetermined light spot shape;

[0008] The objective lens group is used to modulate the spot size of the light beam output by the light source assembly.

[0009] In an optional embodiment of the present application, the light homogenizing and shaping element includes a microlens array arranged on the output light path of the light source assembly, and a condenser lens arranged on the output light path of the microlens array;

[0010] Wherein, the micro-element of the micro-lens array is a micro-element structure of a set shape.

[0011] In an optional embodiment of the present application, the microlens array is a double-row microlens array, and the spacing between the two rows of microlens arrays is equal to the focal length of a single microlens array.

[0012] In an optional embodiment of the present application, the light homogenizing and shaping element includes a light homogenizing rod disposed on the output light path of the light source assembly, and the light emitting end face profile of the light homogenizing rod is a set shape.

[0013] In an optional embodiment of the present application, the light source assembly includes a light source and a collimating lens arranged on an output light path of the light source.

[0014] In an optional embodiment of the present application, the number of the light sources is two, and a collimating lens is arranged on the output light path of each of the light sources; and a light combining element is arranged on the output light path of the two collimating lenses for combining the light beams output by the two collimating lenses for output.

[0015] In an optional embodiment of the present application, the objective lens group includes a doublet lens and a microscope objective lens.

[0016] A microscope device, comprising the illumination system as described in any one of the above items, and an image collector;

[0017] The lighting system is used to project lighting light onto the sample to be tested;

[0018] The image collector is used to collect the image of the sample to be tested under the irradiation of the illumination light.

[0019] In an optional embodiment of the present application, a dichroic mirror is further provided on the output light path of the condenser lens in the illumination system;

[0020] The image collector is arranged on the output light path of the dichroic mirror, and is used for receiving the reflected light from the surface of the sample to be measured that is incident on the image collector through the dichroic mirror.

[0021] In an optional embodiment of the present application, the dichroic mirror is arranged on the optical path between a plurality of lenses included in the objective lens group.

[0022] The present invention provides an illumination system, comprising a light source assembly, a light-homogenizing and shaping element arranged on an output light path of the light source assembly, and an objective lens group arranged on the output light path of the light-homogenizing and shaping element; the light-homogenizing and shaping element is used to homogenize and shape the light emitted by the light source assembly into a light beam with a predetermined light spot shape; the objective lens group is used to modulate the light spot size of the light beam output by the light source assembly.

[0023] In the present application, a uniform light shaping element is used to uniformly shape the output optical path of the light source component into a light beam of a predetermined spot shape, and then the light spot is modulated by the objective lens group to finally make the light beam spot meet the lighting requirements. When the lighting system in the present application is applied to a microscope device, it is only necessary to set the uniform light shaping element to a shape structure that is the same as the effective field of view of the imaging system, so that the light beam spot of the lighting system and the effective field of view of the imaging system can be matched. Compared with the matching of the illumination spot and the effective field of view by limiting the light beam spot shape by an aperture, the energy loss of the light beam can be reduced and the illumination illumination can be improved in the present application; on this basis, because the illumination spot in the present application is a light beam formed after the uniform light beam output by the light source component is uniformly lighted by the uniform light shaping element, the illumination uniformity of the light spot after the mixed output can be guaranteed to a certain extent.

[0024] In summary, when the illumination system in the present application is applied to a microscope device, it can ensure high illumination and good uniformity of the illumination beam on the basis of matching the effective field of view of the imaging system, thereby reducing the noise when collecting and detecting images in the subsequent imaging system, which is beneficial to the accuracy of fluorescent molecule detection.

[0025] The present application also provides a microscope device having the above-mentioned beneficial effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] In order to more clearly illustrate the embodiments of the present invention or the technical solutions of the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0027] Figure 1 A schematic diagram of the optical path structure of the lighting system provided in an embodiment of the present application;

[0028] Figure 2 Schematic diagram of the optical path structure of the microscope device provided in an embodiment of the present application. DETAILED DESCRIPTION

[0029] The illumination system in traditional microscope equipment is a critical illumination system; the critical illumination system directly images the light source on the illuminated sample plane, which makes it difficult to ensure the uniformity of the light spot illumination on the sample plane. For this reason, the Korah illumination system is proposed on this basis. In the Korah illumination system, the Korah mirror is imaged on the sample plane, so that the sample plane is illuminated more evenly.

[0030] However, in order to match the effective field of view of the imaging system, the illumination beam output by the illumination system in the microscope equipment needs to output a light spot with the same shape as the effective field of view of the imaging system. For this reason, it is necessary to use an aperture to shape the light spot shape of the illumination beam. For example, if the effective field of view is rectangular, it is necessary to set an aperture with a rectangular light hole on the optical path of the illumination system so that the light spot shape of the final output illumination beam matches the shape of the effective field of view.

[0031] Obviously, the reason why the aperture can shape the light spot is that it blocks part of the light output by the light source in the lighting system, so that only the light within the rectangular aperture can be output to finally obtain a rectangular light spot. Obviously, this will reduce the power of the lighting beam output by the lighting system to a certain extent.

[0032] To this end, the present application proposes an illumination system that can improve the illumination energy output efficiency of the illumination system and ensure the uniformity of the light spot illumination on the basis of matching the effective field of view of the imaging system, which is beneficial to improving the quality of the image collected and detected by the imaging system.

[0033] In order to enable those skilled in the art to better understand the scheme of the present invention, the present invention is further described in detail below in conjunction with the accompanying drawings and specific implementation methods. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention.

[0034] like Figure 1 As shown, Figure 1 A schematic diagram of the optical path structure of the lighting system provided in an embodiment of the present application. The lighting system may include:

[0035] Light source component 1;

[0036] A light homogenizing and shaping element disposed on the output light path of the light source assembly 1, and an objective lens group 4 disposed on the output light path of the light homogenizing and shaping element;

[0037] The light homogenizing and shaping element is used to homogenize and shape the light emitted by the light source assembly 1 into a light beam with a predetermined light spot shape.

[0038] The objective lens group 4 is used to modulate the spot size of the light beam output by the light source assembly 1 .

[0039] As mentioned above, the lighting system provided in the present application is mainly used in microscope equipment. Based on the requirements for the lighting system in the microscope equipment, the illumination light beam output by the illumination system should match the effective field of view of the imaging system; that is, the shape and size of the spot of the illumination light beam is exactly the same as the shape and size of the effective field of view, so the area where the imaging system can capture and collect the detection image is exactly the area illuminated by the projection spot of the illumination light beam.

[0040] In order to modulate the spot shape of the illumination light beam into the same shape as the effective field of view of the imaging system, a uniform light shaping element is arranged on the output light path of the light source assembly 1 in this embodiment, and the uniform light shaping effect of the uniform light shaping element on the light source assembly 1 is utilized to make the brightness of the light beam spot after passing through the uniform light shaping element uniform, and the spot shape is converted into a spot of a specific shape.

[0041] Optionally, the light homogenizing lens element may be a microlens array 2 disposed on the output light path of the light source assembly 1 and a condenser lens 3 disposed on the output light path of the microlens array 2 .

[0042] The micro-elements of the micro-lens array 2 are micro-element structures of a set shape; and the focusing lens 3 and the objective lens group 4 are used to modulate the spot size of the light beam output by the light source assembly 1 and output it.

[0043] In this embodiment, a microlens array 2 is arranged on the output light path of the light source assembly 1. The micro-elements of the microlens array 2 are used to cut and divide the projected light spot, so that the light spot projected on the microlens array 2 is cut and divided into a plurality of small light spots of the same shape as the micro-elements and transmitted for output. Based on the focusing effect of the condensing lens 3, the light beams corresponding to the small light spots converge, and the small light spots overlap and converge to form an image on the focal plane of the condensing lens 3. Obviously, the light spot on the imaging plane is the same size and shape as a single small light spot, and the illumination is the superposition of the light beams corresponding to all the small light spots.

[0044] It can be determined that the illumination light beam output by the light source assembly 1 can be modulated into a light beam having a spot shape identical to the micro-element shape of the micro-lens array 2 by the modulation of the micro-lens array 2 and the focusing lens 3. Therefore, the micro-lens array 2 in this embodiment can further set the shape of the micro-element so that the shape of each micro-element is identical to the shape of the effective field of view (the size is not necessarily the same, which is equivalent to the proportional scaling shape of the effective field of view); on this basis, the spot size is further modulated by the objective lens group 4 so that the spot shape and size of the illumination light beam output by the illumination system match the effective field of view.

[0045] Based on the above discussion, it can be known that the modulation of the spot shape of the illumination light beam in this embodiment is achieved by cutting and dividing the light beam by the microelements of the microlens array 2 and then re-converging it. In this process, it is not necessary to block and filter out part of the light in the light beam, but the light output by the light source assembly 1 is utilized to the maximum extent. Therefore, on the basis of matching the spot shape with the effective field of view, the utilization rate of the illumination light beam is further improved, ensuring the high illumination output of the illumination light beam.

[0046] Moreover, in the present embodiment, the illumination light beam output by the light source assembly 1 is cut and divided by the microlens array 2 and then overlapped by the focusing lens 3 to form a convergent light beam. Obviously, compared with the illumination light beam directly output by the light source assembly 1, the convergent light beam superimposes the bright and dark parts of different parts of the light spot, thereby reducing the speckle problem of the light beam to a certain extent and improving the illumination uniformity of the light spot corresponding to the illumination light beam.

[0047] It should be noted that the microlens array 2 is generally used in conjunction with a double-row microlens array. Therefore, the microlens array 2 used in this embodiment can also be a double-row microlens array, and the spacing between the two rows of microlens arrays can be equal to the focal length of a single microlens array 2, that is, the micro-element focal length of the microlens array 2; the second row of microlens arrays acts as a field lens to control the divergence angle of the light beam emitted by the first row of microlens arrays.

[0048] The shape of a single microelement is to modulate the light spot to be the same as the effective field of view shape. Therefore, the shape of the microelement should be set based on the effective field of view shape of the imaging system used in the microscope equipment, which can be a rectangle, square, circle, etc.

[0049] Among them, in order to further improve the modulation effect of the microlens array 2 on the illumination light beam, a collimating lens 12 can be set in the light source assembly 1, so that the illumination light beam output by the light-emitting light source 11 in the light source assembly 1 is formed into approximately parallel light to the microlens array 2 through the collimating effect of the collimating lens 12, thereby making the light beam output through the microlens array 2 almost completely parallel light of different field of view angles, so that the light beams corresponding to each small light spot can be better converged into a light spot through the focusing lens 3.

[0050] Furthermore, in order to improve the illumination of the illumination beam output by the light source assembly 1, multiple light sources 11 may be used, each light source 11 is collimated by a collimating lens 12, and then a light combining element 13 such as a dichroic mirror is used to combine the illumination beams output by each light source 11 into a single light beam for output. The wavelengths of the light waves output by each light source 11 may be the same or different.

[0051] The light source 11 in the light source assembly 1 can be an LED light source or a laser light source. When the light source is an LED light source, the spatial light intensity distribution of the output light beam of the LED light source is a Lambertian distribution. In order to achieve the best collimation effect, the collimating lens 12 can be an aspheric surface with NA0.7 to NA0.9, preferably an aspheric surface with NA0.8.

[0052] When the light source 11 is a laser light source, a beam expansion system component can be further arranged on the output light path of the collimating lens 12, so that the light beam output by the laser light source is collimated and expanded before being incident on the microlens array 2. The beam expansion system component can be a Galilean beam expansion system component.

[0053] In addition to the microlens array 2 and the condenser lens 3, the light homogenizing and shaping element may also be an optical element with other structures.

[0054] In another optional embodiment of the present application, the light homogenizing and shaping element includes a light homogenizing rod disposed on the output light path of the light source assembly, and the light emitting end face profile of the light homogenizing rod is a set shape.

[0055] The light incident end of the homogenizing rod can be set close to the light emitting surface of the light source assembly 1. The output light of the light source assembly 1 is parallel light, and there is no need to set a collimating lens. The shape of the light emitting surface of the homogenizing rod can be set to be the same as the shape of the effective field of view.

[0056] The homogenizer rods are divided into hollow and solid ones. The hollow ones are generally coated with a high-reflective film on the inner layer, and the solid ones are generally made of quartz glass or K9 glass. After the light enters the interior of the homogenizer rod from the incident end at a certain divergence angle, it is reflected multiple times. Each reflection will form a virtual light source image. Multiple reflections form a two-dimensional virtual light source matrix, making the light beam evenly distributed, thereby achieving the homogenization effect of the homogenizer rod. In theory, as the length of the homogenizer rod increases, the uniformity of the light beam fluctuates and rises. After reaching a certain extent, continuing to increase the length will not significantly improve the uniformity, but will reduce the brightness. Therefore, in practical applications, the appropriate length of the homogenizer rod can be selected according to actual needs.

[0057] The spot shape of the light beam output by the light homogenizing rod depends on the exit surface of the output light beam. Therefore, in order to obtain a light beam with a specific spot, the shape of the exit surface can be set.

[0058] In addition, in the illumination system provided in the present embodiment, the objective lens group 4 on the output light path of the uniform light shaping element may include a double-cemented lens 41 and a microscope objective lens 42 in the imaging system. The double-cemented lens 41 may also adopt a tube lens structure system consistent with the tube lens of the microscope objective lens 42 in the imaging system, which will not be described in detail in the present application.

[0059] To summarize, the illumination system provided in the present application uses a uniform light shaping element to achieve the shaping of the illumination light beam output by the light source assembly on the basis of minimizing light energy loss, so that the spot shape of the illumination light beam matches the effective field of view of the imaging system, thereby improving the light energy output efficiency of the illumination system, and based on the uniform light effect of the uniform light shaping lens, improving the uniformity of the illumination of the output light spot, ensuring the image quality of the detection images such as fluorescence images collected by the microscope equipment using the illumination system in the present application, reducing image noise, and facilitating improving the accuracy of the detection image analysis results.

[0060] The present application also provides an embodiment of a microscope device, referring to Figure 2 , Figure 2 This is a schematic diagram of the optical path structure of a microscope device provided in an embodiment of the present application. The microscope device may include an illumination system as described in any of the above items, and also includes an image collector 6.

[0061] Obviously, the image collector 6 is also equivalent to an imaging system, which is used to collect the image of the sample plane projected by the illumination system.

[0062] Generally, the illumination system projects illumination light onto the plane where the sample 5 to be tested is located, and the illumination light is reflected from the surface of the sample 5 to be tested and input to the image collector 6. Because the optical path of the illumination light incident on the surface of the sample 5 to be tested and the optical path of the light reflected from the surface of the sample 5 to the surface of the image collector 6 overlap to a certain extent in space, a dichroic mirror 7 can be further arranged on the optical path of the illumination system.

[0063] Reference Figure 2 , the dichroic mirror 7 can be arranged on the output optical path of the uniform light shaping element, taking the uniform light shaping element as the microlens array 2 and the focusing lens 3 as an example. The dichroic mirror 7 is located between the focusing lens 3 and the objective lens group 4, and can also be arranged on the optical path between multiple lenses in the objective lens group 4, for example, between the double glued lens 41 and the microscope objective lens 42. The dichroic mirror 7 reflects the illumination light beam, so that the illumination light beam is incident on the sample to be tested 5 after being deflected by the dichroic mirror 7, and the light reflected from the surface of the sample to be tested 5 is transmitted from the dichroic mirror 7 to the image collector 6; similarly, the dichroic mirror 7 is arranged between the double glued lens 41 and the microscope objective lens 42 to realize light transmission according to the same optical path principle. In actual application, a dichroic mirror 7 can be arranged at any one of the positions between the focusing lens 3 and the objective lens group 4 and between the double glued lens 41 and the microscope objective lens 42. Moreover, in the actual application process, the dichroic mirror 7 can also be used to transmit the illumination light beam and reflect the light reflected from the surface of the sample 5 to realize the technical solution of the present application. Based on the application common sense of the dichroic mirror 7, it can be easily based on Figure 2The optical path structure shown can obtain other optical path structures with similar structures and principles, which will not be described in detail in this application.

[0064] The lighting system in the microscope device of the present application can minimize the light energy loss of the output illumination light, improve the light energy output efficiency, and thus improve the illumination illumination, and can well achieve the mutual matching between the illumination light and the effective field of view of the imaging system, and ensure the uniformity of the illumination beam spot illumination, so that the microscope device can collect and obtain high-quality detection images during practical applications such as fluorescent molecule detection, ensure the accuracy of subsequent image analysis results, and is conducive to the wide application of microscope equipment in fields such as fluorescent molecule detection.

[0065] It should be noted that, in this article, relational terms such as first and second, etc. are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply that there is any such actual relationship or order between these entities or operations. Moreover, the term "include", "comprise" or any other variant thereof is intended to cover non-exclusive inclusion, so that the process, method, article or equipment that includes a series of elements are inherent to the elements. In the absence of more restrictions, the elements limited by the sentence "comprise one..." do not exclude the presence of other identical elements in the process, method, article or equipment that includes the elements. In addition, the above-mentioned technical solution provided in the embodiment of the present application is consistent with the corresponding technical solution in the prior art in principle, and the part is not described in detail, so as not to repeat too much.

[0066] The principles and implementation methods of the present invention are described in this article using specific examples. The description of the above embodiments is only used to help understand the method and core ideas of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made to the present invention without departing from the principles of the present invention, and these improvements and modifications also fall within the scope of protection of the claims of the present invention.

Claims

1. A lighting system, characterized in that: It comprises a light source assembly, a light homogenizing and shaping element arranged on the output light path of the light source assembly, and an objective lens group arranged on the output light path of the light homogenizing and shaping element; The objective lens group is used to modulate the spot size of the light beam output by the light source assembly; Matching the spot shape and size of the illumination light beam output by the illumination system with the effective field of view of the imaging system; The light homogenizing and shaping element comprises a microlens array arranged on the output light path of the light source assembly, and a condenser lens arranged on the output light path of the microlens array; the illumination light beam output by the light source assembly is modulated by the microlens array and the condenser lens to form a light beam with a light spot shape identical to the shape of a microelement of the microlens array; wherein the shape of each microelement of the microlens array is identical to the shape of the effective field of view of the imaging system; The microlens array is a double-row microlens array, and the spacing between the two rows of microlens arrays is equal to the focal length of a single microlens array; The condenser lens is used to overlap and converge a plurality of small light spots with the same shape as the micro-element output by the micro-lens array, so as to form a light spot with the same size and shape as a single small light spot on the focal plane of the condenser lens; The light source assembly comprises a light source and a collimating lens arranged on the output light path of the light source; There are two light sources, and one collimating lens is disposed on the output light path of each light source; and a light combining element is disposed on the output light path of the two collimating lenses for combining the light beams output by the two collimating lenses for output.

2. The lighting system according to claim 1, characterized in that The objective lens group includes a doublet lens and a microscope objective lens.

3. A microscope device, characterized in that: Comprising the lighting system as claimed in claim 1 or 2, and an image collector; The lighting system is used to project lighting light onto the sample to be tested; The image collector is used to collect the image of the sample to be tested under the irradiation of the illumination light.

4. The microscope device according to claim 3, characterized in that A dichroic mirror is also provided on the output light path of the condenser lens in the illumination system; The image collector is arranged on the output light path of the dichroic mirror, and is used for receiving the reflected light from the surface of the sample to be measured that is incident on the image collector through the dichroic mirror.

5. The microscope device according to claim 4, characterized in that The dichroic mirror is arranged on an optical path between a plurality of lenses included in the objective lens group.

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