Optical detection device and sample analyzer

Abstract

The invention discloses an optical detection device and a sample analyzer, in the optical detection device, a bright / dark field illumination assembly is used for emitting an illumination light beam, and a fluorescence excitation assembly is used for emitting an excitation light beam; the bearing table is used for bearing a to-be-detected sample; the infinite objective lens and the barrel lens are used for imaging a to-be-detected sample in a bright field to form a bright field imaging light beam, imaging the to-be-detected sample in a dark field to form a dark field imaging light beam, and performing microscopic imaging on fluorescent particles in the to-be-detected sample to form a fluorescent imaging light beam; the imaging receiving assembly is used for receiving the bright field imaging light beam, the dark field imaging light beam and the fluorescence imaging light beam. The optical detection device is an infinite microscopic optical system, can perform bright / dark field detection on the to-be-detected sample, can perform fluorescence detection on the to-be-detected sample, is compact in structure, high in sharing rate of all optical elements, saves optical element materials, and is low in cost.

Description

Technical Field

[0001] The present application relates to the technical field of medical equipment, and in particular to an optical detection device and a sample analyzer. Background Art

[0002] Blood component testing mainly includes red blood cell testing, white blood cell testing and platelet testing. Blood component testing is an important clinical medical indicator. Optical testing is a common method for testing blood components. Different optical testing devices are generally used for different testing items.

[0003] In the related art, if an optical detection device is to achieve accurate identification, classification and counting of blood cells, it usually requires multiple optical imaging modes to assist each other. The existing optical detection device is not compact enough, has many optical moving parts, and has poor optical performance stability. Summary of the invention

[0004] The present application provides an optical detection device and a sample analyzer to solve the technical problems in the prior art that the optical detection device has a less compact structure, has more optical moving parts, and has poor optical performance stability.

[0005] In order to solve the above technical problems, a technical solution adopted in the present application is: to provide an optical detection device, which includes: a bright / dark field illumination component, a fluorescence excitation component, a carrier platform, an infinite objective lens, a barrel lens and an imaging receiving component, wherein the bright / dark field illumination component is used to emit an illumination beam, and the fluorescence excitation component is used to emit an excitation beam; the carrier platform is located on the light output path of the bright / dark field illumination component and the fluorescence excitation component, and the carrier platform is used to carry a sample to be detected; the infinite objective lens is located on the side of the carrier platform away from the bright / dark field illumination component, the barrel lens is located on the light output path of the infinite objective lens, the image-side focal plane of the infinite objective lens overlaps with the object-side focal plane of the barrel lens, the infinite objective lens and the barrel lens are used to image the sample to be detected in the bright field to form a bright field imaging beam, and to image the sample to be detected in the dark field to form a dark field imaging beam, and to perform microscopic imaging of fluorescent particles in the sample to be detected to form a fluorescence imaging beam; the imaging receiving component is located on the light output path of the barrel lens, and is used to receive the bright field imaging beam, the dark field imaging beam and the fluorescence imaging beam.

[0006] Furthermore, the bright / dark lighting component includes: an illumination light source for emitting an illumination light beam; a first focusing lens, located on the light exit path of the illumination light source, for focusing the illumination light beam; a field diaphragm, located on the light exit path of the first focusing lens; an aperture diaphragm, located on the light exit path of the field diaphragm; and a collimating lens, located on the light exit path of the aperture diaphragm, for collimating the illumination light beam.

[0007] Furthermore, the optical detection device also includes: a dark field condenser, which can be movably placed between the collimating lens and the supporting platform to switch the imaging environment of the sample to be detected from a bright field environment to a dark field environment. In the dark field environment, the dark field condenser is used to block and focus the illumination light beam emitted by the collimating lens to provide a dark field environment for the imaging of the sample to be detected.

[0008] Furthermore, the dark field condenser includes: a dark field ring, arranged on the light output path of the collimating lens, for blocking the illumination light beam within a preset range; a second focusing lens, arranged on the light output path of the dark field ring, for focusing the illumination light beam passing through the dark field ring; and a dark field auxiliary component, arranged on the light output path of the second focusing lens, for focusing the illumination light beam emitted by the second focusing lens to increase the deflection angle of the illumination light beam.

[0009] Furthermore, the dark field auxiliary component includes a third condensing lens, the focal length of the third condensing lens is less than or equal to the focal length of the second condensing lens, or the dark field auxiliary component is a hollow frustum, the inner wall of the dark field auxiliary component is provided with a reflective layer, and the inclination angle of the inner wall of the dark field auxiliary component is greater than the cone angle of the illumination light beam emitted by the second condensing lens.

[0010] Furthermore, the optical detection device also includes: a dark field light shielding sheet, which is movably placed between the infinity objective lens and the imaging receiving assembly to switch the imaging environment of the sample to be detected from a bright field environment to a dark field environment.

[0011] Furthermore, in a dark field environment, the dark field shading sheet is located at the rear focal plane of the infinity objective lens.

[0012] Furthermore, the optical detection device also includes: a dichroic mirror, which is movably arranged so that the dichroic mirror has a first state and a second state. In the first state, the dichroic mirror is located on the light output path of the fluorescent excitation component, and the dichroic mirror is used to emit the excitation light beam to the infinite objective lens, and the infinite objective lens is used to collimate the excitation light beam and then emit it to the sample to be detected. The dichroic mirror is also used to guide the fluorescent imaging light beam emitted by the objective lens to the imaging receiving component; in the second state, the dichroic mirror is not located between the infinite objective lens and the imaging receiving component.

[0013] Furthermore, the optical detection device also includes a light selection component, which includes at least two light selection sub-components connected in sequence, and the at least two light selection sub-components are movably arranged so that at least two light selection sub-components can be selectively moved between the infinity objective lens and the barrel lens to select the imaging mode of the fluorescence imaging beam, the bright field imaging beam and the dark field imaging beam.

[0014] In order to solve the above technical problems, another technical solution adopted in the present application is: to provide a sample analyzer, which includes a sample preparation device and an optical detection device of any of the above embodiments, the sample preparation device is used to prepare the sample to obtain the sample to be detected; the optical detection device is used to perform optical detection on the sample to be detected.

[0015] Beneficial effects of the present application: Different from the prior art, the optical detection device provided by the present application includes: a bright / dark field illumination component and a fluorescence excitation component, a carrier, an infinite objective lens and a tube lens, and an imaging receiving component. The infinite objective lens and the tube lens are used to image the sample to be detected in the bright field to form a bright field imaging beam, to image the sample to be detected in the dark field to form a dark field imaging beam, to perform microscopic imaging of fluorescent particles in the sample to be detected to form a fluorescence imaging beam, and the imaging receiving component is used to receive the bright field imaging beam, the dark field imaging beam, and the fluorescence imaging beam. The optical detection device of the present application uses an infinite objective lens, which will not impose a large restriction on the position of the extended components, such as the adjustable range of the position of the dichroic mirror and the light selection component on one side of the infinite objective lens, and the optical performance stability is good. The present application integrates the fluorescence imaging mode, the bright field imaging mode, and the dark field imaging mode into one, and can perform detection of multiple projects, with clear optical imaging effects and high detection reliability. The imaging systems of multiple optical systems are unified through an infinite objective lens, reducing the cost of setting up a set of optical components for each optical system. The structure is compact, and three optical systems share one imaging system, which can achieve comprehensive detection of samples to be detected, such as blood cells. There is a high sharing rate between optical components, saving optical component materials and low cost. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] In order to more clearly illustrate the technical solutions in the embodiments of the present application, the drawings required for use in the description of the embodiments are briefly introduced below. Obviously, the drawings described below are only some embodiments of the present application. For those skilled in the art, other drawings can be obtained based on these drawings without creative work, among which:

[0017] Figure 1 is a structural schematic diagram of an embodiment of an optical detection device provided by the present application;

[0018] Figure 2 yes Figure 1 A schematic structural diagram of an embodiment of a bright field detection system in an optical detection device shown;

[0019] Figure 3 yes Figure 1 A schematic structural diagram of an embodiment of a dark field detection system in an optical detection device shown;

[0020] Figure 4yes Figure 3 A schematic structural diagram of an embodiment of a dark field condenser in an optical detection device shown;

[0021] Figure 5 yes Figure 4 A schematic structural diagram of an embodiment of a dark field ring in a dark field condenser shown;

[0022] Figure 6 yes Figure 4 A schematic structural diagram of an embodiment of a dark field auxiliary component in a dark field condenser shown;

[0023] Figure 7 yes Figure 4 A schematic structural diagram of another embodiment of a dark field auxiliary component in a dark field condenser shown;

[0024] Figure 8 yes Figure 1 A schematic structural diagram of another embodiment of a dark field detection system in an optical detection device shown;

[0025] Fig. 9 yes Figure 8 A schematic structural diagram of an embodiment of a dark field light shielding sheet in an optical detection device shown;

[0026] Fig.10 yes Figure 1 A schematic structural diagram of an embodiment of a fluorescence detection system in an optical detection device shown;

[0027] Fig.11 It is a structural schematic diagram of an embodiment of a sample analyzer provided by the present application. DETAILED DESCRIPTION

[0028] In order to make the above-mentioned purposes, features and advantages of the present application more obvious and easy to understand, the specific implementation methods of the present application are described in detail below in conjunction with the accompanying drawings. It is to be understood that the specific embodiments described herein are only used to explain the present application, rather than to limit the present application. It should also be noted that, for ease of description, only some structures related to the present application are shown in the accompanying drawings, rather than all structures. Based on the embodiments in the present application, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of this application.

[0029] The terms "first", "second", etc. in this application are used to distinguish different objects, rather than to describe a specific order. In addition, the terms "including" and "having" and any variations thereof are intended to cover non-exclusive inclusions. For example, a process, method, system, product or device that includes a series of steps or units is not limited to the listed steps or units, but optionally includes steps or units that are not listed, or optionally includes other steps or units inherent to these processes, methods, products or devices.

[0030] Reference to "embodiments" herein means that a particular feature, structure, or characteristic described in conjunction with the embodiments may be included in at least one embodiment of the present application. The appearance of the phrase in various locations in the specification does not necessarily refer to the same embodiment, nor is it an independent or alternative embodiment that is mutually exclusive with other embodiments. It is explicitly and implicitly understood by those skilled in the art that the embodiments described herein may be combined with other embodiments.

[0031] The present application provides an optical detection device, which is an infinite microscope optical system. Compared with the finite microscope optical system, the infinite microscope optical system provides an operable space for the structural change of imaging between the infinite objective lens and the barrel lens, and will not impose a large restriction on the position of the extended components, such as the adjustable range of the position of the dichroic mirror and the light selection component on one side of the infinite objective lens, and has good optical performance stability, which has structural advantages. The optical detection device provided by the present application combines the bright field optical detection system, the dark field optical detection system and the fluorescent optical detection system together compactly to realize the free switching of the three imaging modes. Moreover, each imaging module has a high commonality rate, a simple and compact structure, and a low cost.

[0032] See also Figure 1 As shown, Figure 1 1 is a schematic diagram of the structure of an embodiment of the optical detection device provided by the present application. Specifically, the optical detection device 10 includes: a bright / dark field illumination component 11, a fluorescence excitation component 12, a carrier 13, an infinite objective lens 14, a tube lens 15 and an imaging receiving component 16. The bright / dark field illumination component 11 and the fluorescence excitation component 12 are arranged at intervals.

[0033] The bright / dark field illumination component 11 is used to emit an illumination beam, and the fluorescent excitation component 12 is used to emit an excitation beam. The carrier 13 is located on the light output path of the bright / dark field illumination component 11 and the fluorescent excitation component 12. The carrier 13 is used to carry the sample to be detected (not shown in the figure), and the sample to be detected can be a blood sample, a sweat sample, or a saliva sample after a specific treatment. In a specific embodiment, for example, the preparation of a white blood cell sample: the blood and the chemical dye are mixed in a certain proportion for staining, and the staining lasts for 2 minutes, and then 10ul of the mixed solution is sucked with a pipette and dropped on the blood cell counting plate to make the droplets spread evenly. After the diffusion is completed, the sample slide is placed under the bright field optical system for imaging; the preparation of a red blood cell sample: the blood and the red blood cell spheroidization diluent are mixed and diluted in a certain proportion, and the spheroidization treatment is performed for 2 minutes, and then 10ul of the mixed solution is sucked with a pipette and dropped on the blood cell counting plate to make the droplets spread evenly. After the diffusion is completed, the sample slide is placed under the bright field optical system for imaging. Platelet sample preparation: Blood and chemical reagents are mixed and diluted in a certain ratio, treated for 2 minutes, and then 10ul of the mixed solution is pipetted and dropped on the blood cell counting plate to make the droplets spread evenly. After the diffusion is completed, the sample slide is placed under a bright field optical system for imaging. In other embodiments, the sample can also be processed in other ways according to the requirements of the test project to obtain the sample to be tested, such as diluting with a diluent, etc., which are not listed here one by one.

[0034] The illumination beam emitted by the bright / dark field illumination component 11 is emitted to the sample to be detected on the carrier 13, and the sample to be detected can transmit the illumination beam. The excitation beam emitted by the fluorescence excitation component 12 can be emitted to the sample to be detected on the carrier 13, and the fluorescent particles in the sample to be detected can be excited to emit a fluorescent beam under the action of the excitation beam.

[0035] The infinite objective lens 14 is located on the side of the carrier 13 away from the bright / dark field illumination assembly 11, and the lens barrel 15 is located on the light emitting path of the infinite objective lens 14. The image-side focal plane of the infinite objective lens 14 overlaps with the object-side focal plane of the lens barrel 15, and the infinite objective lens 14 is used to magnify and image the sample to be detected. The sample to be detected is imaged at infinity by the infinite objective lens 14, and is imaged on the imaging receiving assembly 16 through the lens barrel 15. The infinite objective lens 14 and the lens barrel 15 are used to image the sample to be detected in the bright field to form a bright field imaging beam, and to image the sample to be detected in the dark field to form a dark field imaging beam, and to perform microscopic imaging of fluorescent particles in the sample to be detected to form a fluorescent imaging beam. The infinite objective lens 14 and the lens barrel 15 can be used for both fluorescence detection and bright / dark field detection, and have a simple and compact structure, a high commonality rate between optical components, and low cost.

[0036] The imaging receiving assembly 16 is located on the light-emitting optical path of the lens barrel lens 15, and is used to receive the bright field imaging light beam, the dark field imaging light beam and the fluorescence imaging light beam. That is, the imaging receiving assembly 16 is used to receive the image of the sample to be detected formed by the infinite objective lens 14 and the lens barrel lens 15. The imaging receiving assembly 16 can be a camera module, for example, the imaging receiving assembly 16 can include a CMOS (Complementary Metal Oxide Semiconductor) camera module, a CMOS camera, or a CCD (charge-coupled device) camera.

[0037] In the present application, a fluorescence detection system is composed of a fluorescence excitation component 12, an objective lens 14, a tube lens 15 and an imaging receiving component 16. The excitation light beam emitted by the fluorescence excitation component 12 is emitted to the sample to be detected, and the sample to be detected is excited to produce fluorescence. The fluorescent particles are imaged by the objective lens 14 and the tube lens 15 to form a fluorescence imaging light beam. The imaging receiving component 16 receives the fluorescence imaging light beam to obtain an image of the sample to be detected, and fluorescence detection and analysis is performed on the sample to be detected based on the imaging.

[0038] The bright / dark field lighting component 11, the objective lens 14, the tube lens 15 and the imaging receiving component 16 constitute a bright / dark field detection system. The illumination light beam emitted by the bright / dark field lighting component 11 is emitted to the sample to be detected. In the bright field, the objective lens 14 and the tube lens 15 image the sample to be detected to form a bright field imaging light beam. In the dark field, the objective lens 14 and the tube lens 15 image the sample to be detected to form a dark field imaging light beam. The imaging receiving component 16 receives the bright / dark field imaging light beam to obtain an image of the sample to be detected, and performs bright / dark field detection and analysis on the sample to be detected based on the imaging.

[0039] Therefore, the optical detection device 10 of the present application uses an infinite objective lens 14, which will not impose a large restriction on the position of the extended components, such as the adjustable range of the position of the dichroic mirror 20 and the light selection component 17 on one side of the infinite objective lens 14 described below, and the optical performance stability is good. The present application integrates the fluorescence detection system and the bright / dark field detection system, which can perform both fluorescence detection and bright / dark field detection on the sample to be detected, so that the optical detection device 10 can perform multiple project detections, thereby improving the scope of application of the optical detection device 10. The cost of setting up a set of optical components for each optical system is reduced, and the structure is compact. Three optical systems share one imaging system, which can realize comprehensive detection of samples to be detected, such as blood cells. There is a high sharing rate between optical components, which saves optical component materials and simplifies the structure of the optical detection device 10. Moreover, the present application adopts an infinite system for imaging, and the imaging effect is clear, which can improve the accuracy of detection of samples to be detected.

[0040] Furthermore, if Figure 1 As shown, the optical detection device 10 further includes a light selection element 17, which is located on the light output path of the infinite objective lens 14, and is used to select imaging modes for the bright field imaging beam, the dark field imaging beam, and the fluorescence imaging beam.

[0041] Specifically, the light selection component 17 may include at least two light selection sub-components (not shown). The light selection component 17 is movably arranged so that one of the at least two light selection sub-components is moved between the infinite objective lens 14 and the tube lens 15 to select an imaging mode. That is, during the movement of the light selection component 17, a suitable light selection sub-component can be selected as needed to select a mode for the imaging light beam.

[0042] like Figure 1 As shown, in this embodiment, multiple light selection components are arranged in a straight line and connected side by side, and the light selection component 17 can move back and forth in the horizontal direction to switch the light selection components. In other embodiments, multiple light selection components can also be arranged in a circle, and the circular light selection component 17 rotates to switch the light selection components.

[0043] exist Figure 1 In the illustrated embodiment, the light selection component 17 includes a light selection sub-component 171, a light selection sub-component 172, a light selection sub-component 173, and a light selection sub-component 174. When the optical detection device 10 performs classification and counting of white blood cells, red blood cells, and platelets on a blood sample, the light selection sub-component 171 may be a green light bandpass filter, the light selection sub-component 172 may be a red light bandpass filter, the light selection sub-component 173 may be a 510nm long pass filter, and the light selection sub-component 174 may be a hollow cavity. For example, when it is necessary to observe red fluorescent cells, the light selection component 17 is moved to switch to the red light bandpass filter, when it is necessary to observe green fluorescent cells, the light selection component 17 is moved to switch to the green light bandpass filter, and when it is necessary to observe red and green fluorescent cells at the same time, the light selection component 17 is moved to switch to the 510nm long pass filter. The filter can filter the fluorescent imaging beam. The cavity is formed with an empty hole (not shown), which can be used to allow the bright field imaging beam or the dark field imaging beam to pass directly. It is understandable that the number and type of filters in the optical selection element 17 can be specifically set according to the requirements of the detection project, and are not specifically limited here.

[0044] Furthermore, if Figure 1As shown, the bright / dark field illumination assembly 11 includes an illumination light source 111, a first focusing lens 112, a field stop 113, an aperture stop 114, and a collimating lens 115. The illumination light source 111 is used to emit an illumination light beam, and the illumination light source 111 can be a white light emitting diode. For example, the illumination light source 111 is a white light emitting diode with a large divergence angle. In other embodiments, the illumination light source 111 can also be a yellow light emitting diode, a red light emitting diode, or a green light emitting diode, etc.

[0045] The first focusing lens 112 is located on the light-emitting path of the illumination light source 111, and is used to focus the illumination light beam emitted by the illumination light source 111. The field diaphragm 113 is provided on the light-emitting path of the first focusing lens 112, and the field diaphragm 113 is used to perform beam reduction processing on the illumination light beam to control the object side field of view. The field diaphragm 113 is used to adjust the size of the object side field of view illumination surface of the imaging system, and its light transmission diameter can be 1.5 mm, 2 mm, 3 mm or 4 mm. The aperture diaphragm 114 is located on the light-emitting path of the field diaphragm 113, and the aperture diaphragm 114 is used to filter out stray light and adjust the imaging contrast of the optical detection device 10, and the light transmission diameter of the aperture diaphragm is 1.5 mm, 2 mm, 3 mm, or 4 mm. The collimating lens 115 is located on the light-emitting path of the aperture diaphragm 114, and is used to collimate the illumination light beam. In the optical detection device 10 of the present application, a position for a dark field device is reserved behind the collimating lens 115, which provides conditions for the optical detection device 10 to switch to dark field imaging.

[0046] like Figure 2 As shown, in the bright field, the illumination light beam emitted by the illumination light source 111 is converged and focused by the first focusing lens 112, and then is reduced by the field diaphragm 113 to control the object field of view. The reduced illumination light beam passes through the aperture diaphragm 114 to filter out stray light and adjust the imaging contrast of the optical detection device 10. The illumination light beam passing through the aperture diaphragm 114 is collimated by the collimating lens 115 and emitted to the sample to be detected to achieve uniform illumination. That is, the illumination light source 111, the first focusing lens 112, the field diaphragm 113, the aperture diaphragm 114, and the collimating lens 115 constitute a bright field illumination system. The bright field illumination system, the infinite objective lens 14, the tube lens 15, and the imaging receiving assembly 16 constitute a bright field detection system. During bright / dark field detection, the light selection element 17 can be switched to an empty hole, or the light selection element 17 can be pulled out to allow the bright field imaging light beam or the dark field imaging light beam to pass directly.

[0047] Furthermore, a dark field device is added to the bright field detection system to switch the optical detection device 10 to a dark field detection system. In other words, the dark field detection system shares the main structure of the bright field detection system, specifically consisting of the bright field detection system and the dark field device. Figure 3 As shown, a dark field condenser 18 can be added to the bright field detection system to form a dark field detection system.

[0048] Specifically, Figure 3 As shown, the dark field condenser 18 is movably placed between the collimating lens 115 and the carrier 13 to switch the imaging environment of the sample to be detected from the bright field environment to the dark field environment. That is, when the optical detection device 10 is switched to the dark field environment, the dark field condenser 18 is placed between the collimating lens 115 and the carrier 13. The dark field condenser 18 is used to converge the illumination light beam emitted by the collimating lens 115 so that the illumination light beam is focused on the sample to be detected, and the scattered light cannot enter the entrance pupil of the infinite objective lens 14, providing a dark field environment for the imaging of the sample to be detected. When the optical detection device 10 is switched to the bright field environment, the dark field condenser 18 is not located between the collimating lens 115 and the carrier 13.

[0049] Furthermore, if Figure 4 As shown, the dark field condenser 18 includes a dark field ring 181, a second condensing lens 182 and a dark field auxiliary member 183. The dark field ring 181 is arranged on the light output path of the collimating lens 115. Figure 5 As shown, Figure 5 yes Figure 4 The structure schematic diagram of an embodiment of a dark field ring in a dark field condenser shown in FIG. 1 , the dark field ring 181 is used to block the illumination light beam within a preset range. The second condensing lens 182 is arranged on the light exiting path of the dark field ring 181, and is used to focus the illumination light beam emitted by the dark field ring 181. The dark field auxiliary member 183 is arranged on the light exiting path of the second condensing lens 182, and is used to increase the degree of deflection of the illumination light beam emitted by the second condensing lens 182, thereby increasing the numerical aperture of the dark field illumination system.

[0050] In the dark field optical path, the illumination light beam coming out of the collimating lens 115 passes through the dark field ring 181, is focused by the second condensing lens 182, and is further deflected by the dark field auxiliary component 183, so that the illumination light beam is focused on the sample to be detected, providing a dark field environment for the dark field imaging of the sample to be detected, and the scattered light of the illuminated sample to be detected is received and imaged via the infinity objective lens 14 and the tube lens 15 to form a dark field imaging light beam, and the imaging receiving component 16 receives the dark field imaging light beam to observe the sample to be detected. For example, when observing blood cells, the imaging background is dark, and only the blood cells are imaged as bright spots. The numerical aperture of the dark field illumination system of the present application is relatively large. Therefore, it can match the dark field imaging of the infinity objective lens 14 with a larger numerical aperture, thereby improving the clarity of the imaging.

[0051] In some specific embodiments, Figure 6 As shown, Figure 6 yes Figure 4Schematic structural diagram of an embodiment of a dark-field auxiliary member in the shown dark-field condenser. The dark-field auxiliary member 183 may include a third condenser lens disposed on the light-emitting optical path of the second condenser lens 182, and the focal length of the third condenser lens is less than or equal to the focal length of the second condenser lens 182. The third condenser lens is used to increase the deflection degree of the illumination light beam emitted by the second condenser lens 182, thereby increasing the numerical aperture of the dark-field illumination system. For example, by adding a third condenser lens with a numerical aperture of 0.5 behind the second condenser lens 182, the deflection degree of the dark-field emitted light is increased, and thus the numerical aperture of the dark-field illumination system can reach 0.5. By the above method, the numerical aperture of the dark-field illumination system can be increased. Therefore, it can match the dark-field imaging of an infinity objective 14 with a larger numerical aperture and improve the imaging clarity.

[0052] Optionally, in some other embodiments, such as Figure 7 shown, the dark-field auxiliary member 183 may include a dark-field condenser with a hollow (hollow) structure, and a reflective layer is provided on the inner wall of the dark-field condenser. The inclination angle α of the inner wall of the dark-field condenser is greater than the cone angle of the illumination light beam after being condensed by the second condenser lens 182. Specifically, the size of the inner diameter of the dark-field condenser gradually decreases in the direction approaching the carrier 13. The inclination angle of the inner wall of the dark-field condenser refers to the angle between the inner wall of the dark-field condenser and the central axis of the dark-field condenser, such as Figure 7 the angle α marked in. The cone angle of the illumination light beam after being condensed by the second condenser lens 182 refers to the angle between the illumination light beam after being condensed by the second condenser lens 182 and the central axis of the illumination light beam.

[0053] Furthermore, the dark-field auxiliary member 183 may be in the shape of a hollow frustum, and the cone angle (inclination angle) of the inner side wall of the frustum is greater than the cone angle of the light beam after being condensed by the second condenser lens 182. Among them, the greater the inclination angle of the dark-field condenser, the stronger the refractive ability, and the larger the numerical aperture of the entire dark-field illumination system. This embodiment can achieve dark-field imaging with the numerical aperture NA of the infinity objective 14 in the range of 0 < NA < 0.75. For example, when the frustum cone angle reaches 70 degrees - 80 degrees, the numerical aperture of the dark-field illumination system can reach 0.8, and the range of the numerical aperture NA of the infinity objective 14 that can be matched is: 0 < NA < 0.75.

[0054] In the dark-field detection system, the illumination light source 111, the first focusing lens 112, the field stop 113, the aperture stop 114, the collimating lens 115, the dark-field condenser 18, the infinity objective 14, the tube lens 15, and the imaging receiving component 16 can all be coaxially placed to improve the utilization rate of light.

[0055] In the above embodiment, it is only necessary to add a dark field condenser 18 after the bright field illumination system and adjust the height of the dark field illumination system so that the dark field condenser 18 focuses the illumination beam on the sample to be detected. That is, the sample slide on the carrier 13 is located at the focal plane of the dark field auxiliary component 183, and the position of the focal plane is the object plane of the infinite objective lens 14, providing a dark field environment for imaging the sample to be detected. This optimized dark field can match the dark field imaging of the infinite objective lens 14 with a larger numerical aperture, providing conditions for clear dark field imaging of the sample to be detected.

[0056] Alternatively, if Figure 8 and Fig. 9 As shown, the dark field detection system may further include a bright field detection system and a dark field shading sheet 19, and the dark field shading sheet 19 may be movably placed between the infinite objective lens 14 and the imaging receiving assembly 16 to switch the imaging environment of the sample to be detected from the bright field environment to the dark field environment. When the imaging environment of the sample to be detected is a dark field environment, the dark field shading sheet 19 is arranged on the light exit path of the infinite objective lens 14. Specifically, in the dark field environment, the dark field shading sheet 19 may be located on the back focal plane of the infinite objective lens 14. When the imaging environment of the sample to be detected is switched to a bright field environment, the dark field shading sheet 19 is not located between the infinite objective lens 14 and the imaging receiving assembly 16.

[0057] The dark field imaging method in this embodiment is as follows: the parallel light emitted by the bright / dark field illumination component 11 is projected onto the sample to be detected on the carrier 13, the light passing through the sample to be detected is focused by the infinite objective lens 14 onto the back focal plane of the infinite objective lens 14, a dark field light shielding sheet 19 is inserted into the back focal plane of the infinite objective lens 14 to block the focused light spot, and the illuminated sample to be detected is imaged at infinity by the infinite objective lens 14, and is imaged by the tube lens 15 onto the imaging receiving component 16. At this time, only the image is lit and the background is dark, so dark field imaging can be achieved.

[0058] In the dark field detection system of this embodiment, the illumination light source 111, the first focusing lens 112, the field diaphragm 113, the aperture diaphragm 114, the collimating lens 115, the infinity objective lens 14, the dark field shading sheet 19, the tube lens 15 and the imaging receiving assembly 16 can all be coaxially placed to improve the utilization rate of light.

[0059] The shading diameter of the dark field shading sheet 19 has a one-to-one correspondence with the illumination area of ​​the bright field illumination system on the sample to be detected, and the shading diameter may be 2 mm, 2.5 mm or 3 mm, etc.

[0060] In this embodiment, only a dark field shielding sheet is added to the rear focal plane of the infinite objective lens 14 in the bright field imaging system, and the bright field illumination system remains unchanged, so that the dark field mode can be realized. The bright / dark field detection system switching method is simple and the cost is low.

[0061] Furthermore, if Figure 1 and Fig.10 As shown, the optical detection device 10 may further include: a dichroic mirror 20. The dichroic mirror 20 is movably arranged so that the dichroic mirror 20 has a first state and a second state.

[0062] In the first state, the dichroic mirror 20 is located on the light output path of the fluorescence excitation component 12, and the dichroic mirror 20 is used to guide the excitation light beam to the infinite objective lens 14, and the infinite objective lens 14 is used to collimate the excitation light beam and then emit it to the sample to be detected, that is, the excitation light beam is emitted from the infinite objective lens 14 as collimated light, so as to achieve uniform excitation, thereby improving the accuracy of sample detection. The dichroic mirror 20 can also reflect the excitation light beam into the infinite objective lens 14, and the dichroic mirror 20 can allow the fluorescence imaging light beam to pass through the lens barrel lens 15. It can be understood that in other embodiments, the dichroic mirror 20 can also reflect the fluorescence imaging light beam to the lens barrel lens 15 and pass the laser light beam through the infinite objective lens 14. In the above manner, the structure of the optical detection device 10 can be made more compact.

[0063] In the second state, the dichroic mirror 20 is not located between the infinity objective lens 14 and the imaging receiving assembly 16. That is, in the bright field detection mode and the dark field detection mode, the dichroic mirror 20 can be withdrawn to prevent the dichroic mirror 20 from affecting the bright field imaging beam and the dark field imaging beam, thereby affecting the accuracy of the bright field detection and the dark field detection.

[0064] The fluorescence excitation component 12, the dichroic mirror 20 and the infinite objective lens 14 constitute a fluorescence excitation system. Specifically, the fluorescence excitation component 12 includes: an excitation light source 121, an aperture 122, a fourth condensing lens 123 and a filter 124 arranged in sequence. The excitation light source 121 is used to emit an excitation light beam. For example, the excitation light source 121 can be a blue light emitting diode. After the excitation light beam passes through the aperture 122 to filter out stray light, the excitation light beam is converged to the back focal plane of the infinite objective lens 14 through the fourth condensing lens 123, wherein the excitation light beam is reflected by the filter 124 and the dichroic mirror 20 before being converged to the infinite objective lens 14, and the excitation light beam converged on the back focal plane of the infinite objective lens 14 is collimated and emitted by the infinite objective lens 14 to achieve uniform excitation.

[0065] Furthermore, in the fluorescence excitation component 12, the geometric diameters of the aperture 123, the second condensing lens 122, and the filter 124 are the same, and the excitation light source 121, the aperture 122, the second condensing lens 123, the filter 124, the dichroic mirror 20 and the infinity objective lens 14 can be coaxially placed to improve the utilization of light.

[0066] The preparation process of the sample to be tested for fluorescence detection can be: the sample to be tested is mixed with the fluorescent dye in a certain proportion for staining. Leukocyte sample preparation: blood and fluorescent dye are mixed in a certain proportion for staining, staining for 2 minutes, then 10ul of the mixed solution is sucked with a pipette and dropped on the blood cell counting plate to make the droplets spread evenly. After the diffusion is completed, the sample slide is placed under the fluorescence optical system for imaging; platelet sample preparation: blood and fluorescent dye are mixed in a certain proportion for staining, processing for 2 minutes, then 10ul of the mixed solution is sucked with a pipette and dropped on the blood cell counting plate to make the droplets spread evenly, and after the diffusion is completed, the sample slide is placed under the fluorescence system for imaging.

[0067] The principle of fluorescence detection imaging is as follows: the sample to be detected is imaged on the imaging receiving component 16 by the infinite objective lens 14 and the tube lens 15. During the imaging process, the light selection component 17 switches to a suitable light selection sub-component, and the fluorescence excited by the sample to be detected is selected again, and unnecessary light is filtered out to achieve fluorescence imaging of the sample to be detected.

[0068] In the fluorescence imaging mode, the optical selector 17 can be switched to a green bandpass filter to realize a fluorescent green light mode to observe blood cells containing DNA (DeoxyriboNucleic Acid); in the fluorescence imaging mode, it can be switched to a red bandpass filter to realize a fluorescent red light mode to observe blood cells containing RNA (Ribonucleic acid); in the fluorescence imaging mode, it can be switched to a 510nm longpass filter to realize a fluorescent red-green light mode, and blood cells containing DNA and RNA can be observed simultaneously.

[0069] The optical detection device 10 of the present application integrates the fluorescence imaging mode, the bright field imaging mode, and the dark field imaging mode into one, and the fluorescence imaging mode and the bright / dark field optical imaging mode share the main imaging system. The bright and dark field optical imaging modes share the main lighting system and the imaging system, and have a compact structure, a high sharing rate between the optical components, saving optical component materials, low cost, and easy disassembly. The optical detection device 10 can realize the free switching of the fluorescence imaging mode, the bright field imaging mode, and the dark field imaging mode, with clear imaging and high resolution, providing a strong platform support for the identification, classification, and counting of the samples to be detected.

[0070] The present application also provides a sample analyzer, such as Fig.11 As shown, Fig.11 It is a structural schematic diagram of an embodiment of a sample analyzer provided in the present application. Specifically, the sample analyzer 30 includes: a sample preparation device 31 and an optical detection device 32. The sample preparation device 31 is used to prepare the sample to obtain a sample to be detected, and the optical detection device 32 is used to perform optical detection on the sample to be detected.

[0071] The optical detection device 32 of this embodiment may be the optical detection device 10 of any of the above embodiments. For the structure of the optical detection device 32 , please refer to the introduction of any of the above embodiments, which will not be described again.

[0072] The sample analyzer 30 in the present application may be a blood analyzer or a body fluid analyzer, etc. The test sample of the sample analyzer 30 may be blood, such as animal blood or human blood, etc., and the test sample may also be body fluids such as saliva, sweat or urine. The sample analyzer 30 of the present application can perform multiple tests on the sample, has a wide range of applications, and has a simple structure and low cost.

[0073] The above description is only an implementation method of the present application, and does not limit the patent scope of the present application. Any equivalent structure or equivalent process transformation made using the contents of the description and drawings of this application, or directly or indirectly applied in other related technical fields, are also included in the patent protection scope of the present application.

Claims

1. An optical detection device, characterized in that: The optical detection device comprises: A bright / dark field illumination component and a fluorescence excitation component, wherein the bright / dark field illumination component is used to emit an illumination beam, and the fluorescence excitation component is used to emit an excitation beam; A carrying platform, located on the light output path of the bright / dark lighting component and the fluorescent excitation component, and the carrying platform is used to carry the sample to be detected; An infinite objective lens and a tube lens, wherein the infinite objective lens is located at a side of the carrier platform away from the bright / dark illumination assembly, the tube lens is located on a light-emitting optical path of the infinite objective lens, an image-side focal plane of the infinite objective lens overlaps with an object-side focal plane of the tube lens, the infinite objective lens and the tube lens are used to image the sample to be detected in a bright field to form a bright-field imaging beam, and to image the sample to be detected in a dark field to form a dark-field imaging beam, and to perform microscopic imaging of fluorescent particles in the sample to be detected to form a fluorescent imaging beam; The imaging receiving component is located on the light output path of the lens barrel and is used for receiving the bright field imaging light beam, the dark field imaging light beam and the fluorescent imaging light beam.

2. The optical detection device according to claim 1, characterized in that: The bright / dark lighting assembly comprises: An illumination light source, used for emitting the illumination light beam; A first focusing lens, located on the light output path of the illumination light source, and used for focusing the illumination light beam; A field stop, located on the light-outgoing path of the first focusing lens; An aperture stop, located on the light exit path of the field stop; A collimating lens is located on the light-emitting path of the aperture diaphragm and is used to collimate the illumination light beam.

3. The optical detection device according to claim 2, characterized in that: The optical detection device also includes: A dark field condenser is movably placed between the collimating lens and the supporting platform to switch the imaging environment of the sample to be detected from a bright field environment to a dark field environment. In the dark field environment, the dark field condenser is used to block and focus the illumination light beam emitted by the collimating lens to provide a dark field environment for the imaging of the sample to be detected.

4. The optical detection device according to claim 3, characterized in that: The dark field condenser comprises: A dark field ring, arranged on the light output path of the collimating lens, for shielding the illumination light beam within a preset range; A second focusing lens is provided on the light-emitting path of the dark field ring and is used to focus the illumination light beam passing through the dark field ring; The dark field auxiliary component is arranged on the light output path of the second focusing lens, and is used for focusing the illumination light beam emitted by the second focusing lens to increase the deflection angle of the illumination light beam.

5. The optical detection device according to claim 4, characterized in that: The dark field auxiliary comprises a third condenser lens, the focal length of the third condenser lens is less than or equal to the focal length of the second condenser lens, or, The dark field auxiliary component is in the shape of a hollow truncated cone, and a reflective layer is provided on the inner wall of the dark field auxiliary component. The inclination angle of the inner wall of the dark field auxiliary component is greater than the cone angle of the illumination light beam emitted by the second condenser lens.

6. The optical detection device according to claim 1, characterized in that: The optical detection device also includes: The dark field light shielding sheet is movably placed between the infinity objective lens and the imaging receiving assembly to switch the imaging environment of the sample to be detected from a bright field environment to a dark field environment.

7. The optical detection device according to claim 6, characterized in that: In the dark field environment, the dark field shading sheet is located at the rear focal plane of the infinity objective lens.

8. The optical detection device according to claim 1, characterized in that: The optical detection device further comprises: a dichroic mirror, the dichroic mirror being movably arranged so that the dichroic mirror has a first state and a second state, In the first state, the dichroic mirror is located on the light output path of the fluorescence excitation component, the dichroic mirror is used to emit the excitation light beam to the infinite objective lens, the infinite objective lens is used to collimate the excitation light beam and then emit it to the sample to be detected, and the dichroic mirror is also used to guide the fluorescence imaging light beam emitted by the objective lens to the imaging receiving component; In the second state, the dichroic mirror is not located between the infinity objective lens and the image receiving assembly.

9. The optical detection device according to claim 1, characterized in that: The optical detection device further comprises a light selection element, The light selection component includes at least two light selection sub-components connected in sequence, and the at least two light selection sub-components are movably arranged so that one of the at least two light selection sub-components can be moved between the infinity objective lens and the tube lens to select the imaging mode of the fluorescence imaging beam, the bright field imaging beam and the dark field imaging beam.

10. A sample analyzer, characterized in that: The sample analyzer comprises: A sample preparation device, used for preparing samples to obtain samples to be tested; The optical detection device according to any one of claims 1 to 9 is used to perform optical detection on the sample to be detected.

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