CN211878296U - Micro-imaging module of miniature fluorescence - Google Patents
Micro-imaging module of miniature fluorescence Download PDFInfo
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Abstract
The utility model discloses a micro fluorescence microscopic imaging module, which comprises an imaging lens module and a lighting device; the imaging lens module comprises an objective lens and a cylindrical lens which are symmetrically arranged in sequence from an object space to an image space, an image sensor with high spatial sampling rate arranged at the image side of the cylindrical lens, and an emission optical filter arranged in front of the objective lens, or between the objective lens and the cylindrical lens, or between the cylindrical lens and the image sensor; the illuminating device and the imaging lens module are separately arranged to form uniform illumination on the sample; the excitation light path of the lighting device is separated from the detection light path of the imaging lens module. The utility model discloses an approximate symmetrical structure designs objective and section of thick bamboo mirror, combines the sensor of high spatial sampling rate, realizes the high resolution formation of image under low magnification. The design can effectively shorten the conjugate distance and solve the problem that the image distance is still long after the microscope is miniaturized. The three excitation lighting schemes save the arrangement of adding an excitation filter and a dichroic mirror, and can meet the lighting requirements of different observation samples.
Description
Technical Field
The utility model relates to an optical imaging field especially relates to a micro-imaging module of miniature fluorescence.
Background
In the field of life sciences, fluorescence microscopy is often used. Fluorescence microscopes typically include a light source, excitation filters, dichroic mirrors, objective lenses, emission filters, tube lenses, detectors, and the like. The light source obtains exciting light through the exciting filter, and the exciting light is reflected by the dichroic mirror to enter the objective lens and is focused on the surface of the sample. The excited fluorescence signal is collected by the objective lens, passes through the dichroic mirror and the optical filter, and is imaged to the target surface of the detector by the cylindrical mirror. Such microscopes often have a certain magnification, which inevitably results in short object distance and long image distance, and the image space must be left with enough space to meet the requirements of the optical path arrangement. In addition, the light source needs to be connected with the commercial power, the detector such as a CCD or a CMOS also needs to be connected with the commercial power or a computer for power supply, and the display and the storage of the image also need to be connected with the computer. Moreover, the microscope must be provided with a focusing mechanism, and the mechanical structure of the microscope is complex and occupies a small space no matter the microscope is manually focused or automatically focused. For these reasons, fluorescence microscopes are generally bulky, complex, expensive, and not portable.
The core components of the fluorescence microscope capable of imaging are an objective lens and a tube lens, and the main factor for limiting the miniaturization of the fluorescence microscope is that the system has larger magnification, so that the focal length of the tube lens is usually very long. And the filter set continues to shrink with previous settings without further consideration of its miniaturized design.
SUMMERY OF THE UTILITY MODEL
The to-be-solved technical problem of the utility model lies in that, to prior art's defect, a miniature fluorescence microscopic imaging module is provided.
The utility model provides a technical scheme that its technical problem adopted is: constructing a micro fluorescent microscopic imaging module which comprises an imaging lens module and an illuminating device;
the imaging lens module comprises an objective lens and a tube lens which are symmetrically arranged in sequence from an object space to an image space, an image sensor with high spatial sampling rate and arranged at the image side of the tube lens, and an emission optical filter arranged in front of the objective lens, or between the objective lens and the tube lens, or between the tube lens and the image sensor;
the illuminating device and the imaging lens module are arranged separately to form uniform illumination on the sample;
an excitation light path of the illuminating device is separated from a detection light path of the imaging lens module; the detection light path comprises a sample, an emission optical filter, an objective lens, a cylindrical lens and an image sensor from an object side to an image side in sequence; or a sample, an objective lens, an emission filter, a barrel mirror, and an image sensor; or the sample, the objective lens, the barrel mirror, the emission filter and the image sensor.
Preferably, in the micro fluorescence microscopic imaging module of the present invention, the objective lens and the lens assembly is composed of a lens group, a distance from an object surface to an image surface of the lens assembly on an optical axis is TTL, a distance from an object side surface to an image side surface of the lens assembly on the optical axis is TD, which satisfies the following relationship: TD/TTL is more than or equal to 0.15 and less than or equal to 0.9;
focal length f of the objective lens1And focal length f of said barrel mirror2Satisfies the following relationship: f is not less than 0.11/f2≤10；
The optical cylinder length of the lens group satisfies the following relationship: TTL/(f) is not less than 0.21+f2)≤15；
Distance L from object space main plane of objective lens to object planeObjThe following relationship is satisfied: 0.5f1≤LObj≤1.5f1；
The cylindrical mirrorIs located at a distance L from the image-side principal plane to the image planeImaSatisfies the following relation: 0.5f2≤LIma≤1.5f2。
Preferably, in the micro fluorescence microscopic imaging module of the present invention, the illumination device includes an excitation light source disposed at the periphery of the objective lens, and a uniform light guide plate corresponding to the excitation light source;
and the light emitted by the excitation light source forms uniform illumination on the sample after passing through the uniform light guide plate.
Preferably, in the micro fluorescence microscopic imaging module of the present invention, the excitation light source includes a plurality of excitation light sources, and the excitation light sources are uniformly distributed on the periphery of the objective lens in an annular shape; the light homogenizing light guide plate is an annular light guide plate arranged in the light outgoing direction of the excitation light source; and the end face of the annular light guide plate is substantially flush with the end face of the objective lens close to the sample.
Preferably, in the micro fluorescence microscopic imaging module of the present invention, the illumination device includes an excitation light source disposed on one side of the sample; the light emitted by the excitation light source irradiates a target area from the side of the sample.
Preferably, in the micro fluorescence micro imaging module of the present invention, the micro fluorescence micro imaging module further includes a support arm disposed at one side of the imaging lens module; the excitation light source is installed on the supporting arm.
Preferably, in the micro fluorescence microscopic imaging module of the present invention, the lighting device set up in the sample below, including corresponding set up in first excitation light source under the objective visual field periphery, with first even light guide plate that first excitation light source corresponds and corresponding to the shielding plate that objective visual field edge set up, light follows the sample side below shines to the target area.
Preferably, in the micro fluorescence microscopic imaging module of the present invention, the illumination device further includes a second excitation light source correspondingly disposed under the field of view of the objective lens, and a second uniform light guide plate corresponding to the second excitation light source, and the light is irradiated to the target area from right below the sample;
the shielding plate isolates the first excitation light source and the second excitation light source, and the first uniform light guide plate and the second uniform light guide plate respectively.
Preferably, in the micro fluorescence microscopic imaging module of the present invention, the micro fluorescence microscopic imaging module further includes a support base, the support base includes a support arm and a base fixed at the end of the support arm; the support arm fixes the specimen from the side thereof.
Preferably, in the micro fluorescence microscopic imaging module of the present invention, the excitation light source is a laser diode or a light emitting diode;
the objective lens and the tube lens are respectively composed of at least three lens.
Preferably, in the micro fluorescence microscopic imaging module of the present invention, the micro fluorescence microscopic imaging module further includes a driving circuit board with a USB interface, for driving the lighting device and the image sensor; andor or
And the protective glass is arranged in front of the objective lens.
Preferably, in the micro fluorescence microscopic imaging module of the present invention, the micro fluorescence microscopic imaging module further includes a focusing motor, which drives the objective lens or the barrel lens or the whole movement composed of the objective lens, the emission filter and the barrel lens to realize focusing.
Through implementing the utility model discloses, following beneficial effect has:
the utility model discloses based on traditional infinity rectifies fluorescence microscope's structure, adopts approximate symmetrical structure design objective and section of thick bamboo mirror, combines the sensor of high spatial sampling rate, realizes the high resolution formation of image under low magnification. The design can effectively shorten the conjugate distance and solve the problem that the image distance is still long after the microscope is miniaturized. The three excitation lighting schemes save the arrangement of adding an excitation filter and a dichroic mirror, and can meet the lighting requirements of different observation samples. The three different positions of the emission filter reduce aberration and ensure imaging quality. In addition, due to the consideration of convenient carrying and simple operation, the driving circuit of the whole microscope can supply power only through simple USB connection, and the image of the image sensor can be connected to a computer and a mobile phone end through the USB for displaying and storing. In addition, this microscopic imaging module can also add the focusing motor, because the small of objective and section of thick bamboo mirror, light in weight uses ripe focusing motor such as voice coil motor alright with drive the focusing of lens group.
Drawings
The invention will be further explained with reference to the drawings and examples, wherein:
FIG. 1 is a schematic structural diagram of a first embodiment of a micro fluorescence microscopy imaging module according to the present invention;
FIG. 2 is a schematic structural diagram of a second embodiment of the micro fluorescence microscopy imaging module of the present invention;
FIG. 3 is a schematic structural diagram of a third embodiment of the fluorescence microscopy imaging module of the present invention;
fig. 4 is a schematic structural diagram of a fourth embodiment of the micro fluorescence microscopic imaging module of the present invention.
Detailed Description
In order to clearly understand the technical features, objects, and effects of the present invention, embodiments of the present invention will be described in detail with reference to the accompanying drawings.
It should be understood that the directions and positional relationships indicated by the terms "front", "back", "upper", "lower", and the like are constructed and operated in specific directions based on the directions and positional relationships shown in the drawings, and are only for convenience of describing the present invention, and do not indicate that the device or element indicated must have a specific direction, and thus, should not be construed as limiting the present invention.
As shown in fig. 1-4, the present invention constructs a micro fluorescence microscopic imaging module, which comprises an imaging lens module and an illumination device;
the imaging lens module comprises an objective lens 18 and a tube lens 16 which are symmetrically arranged from an object side to an image side in sequence, an image sensor 28 which is arranged on the image side of the tube lens 16 and has a high spatial sampling rate, and an emission filter 17 which is arranged in front of the objective lens 18, or between the objective lens 18 and the tube lens 16, or between the tube lens 16 and the image sensor 28. The illumination device is disposed separately from the imaging lens module to provide uniform illumination of the sample 10. An excitation light path of the illuminating device and a detection light path of the imaging lens module are separately arranged; the detection light path comprises a sample 10, an emission filter 17, an objective lens 18, a cylindrical lens 16 and an image sensor 28 from an object side to an image side in sequence; or the sample 10, the objective lens 18, the emission filter 17, the barrel mirror 16 and the image sensor 28; or sample 10, objective lens 18, barrel mirror 16, emission filter 17 and image sensor 28.
In some embodiments, the objective lens 18 and the tube lens 16 are composed of at least three lenses, the lens material can be glass or plastic, and the surface shape of the lens can be set to be spherical or aspherical according to the requirement of aberration correction. The objective lens 18 and the cylindrical lens 16 constitute a lens group which is an infinity corrected microscope, adopts an approximately symmetrical design, facilitates optimization of aberration, and can realize a resolution of 1 μm at low magnification. The low magnification can effectively reduce the image distance length, realize the miniaturization of the microscope and increase the field range.
Wherein, an axial distance from an object plane to an image plane of the lens assembly is TTL, and an axial distance from an object-side surface to an image-side surface of the lens assembly is TD, which satisfy the following relationships: TD/TTL is more than or equal to 0.15 and less than or equal to 0.9;
focal length f of objective lens 181And focal length f of the barrel mirror 162Satisfies the following relationship: f is not less than 0.11/f2≤10；
The optical cylinder length of the lens group satisfies the following relationship: TTL/(f) is not less than 0.21+f2)≤15；
Distance L from the object main plane of the objective 18 to the object planeObjThe following relationship is satisfied: 0.5f1≤LObj≤1.5f1；
Distance L from image side principal plane to image side of cylindrical mirror 16ImaSatisfies the following relation: 0.5f2≤LIma≤1.5f2。
In some embodiments, the emission filter 17 may be an absorption filter or an interference filter, which, when placed in front of the objective lens 18, may reduce the distance between the objective lens 18 and the tube lens 16, facilitate pupil articulation, and better correct aberrations; when it is arranged between the objective lens 18 and the tube lens 16, because the light incident to the optical filter is parallel light, the aberration can be avoided; when the optical fiber is arranged between the cylindrical mirror 16 and the image sensor 28, the incident angle of light is small when the optical fiber works in an afocal mode, and the function of filtering light is favorably realized.
In some embodiments, the micro fluorescence microscopy imaging module further comprises a driving circuit board 11 with a USB interface for driving the illumination means, the image sensor 28 and the focus motor 27. The USB interface can be connected with equipment such as a computer, a mobile phone and the like to realize power supply and display images.
In some embodiments, the illumination device includes an excitation light source 19 and a dodging light guide plate 29 corresponding to the excitation light source 19. The excitation light source 19 may be a laser diode or a light emitting diode, and the light-homogenizing light guide plate 29 may be made of PC (polycarbonate), PMMA (acrylic), glass, or the like, and the light-homogenizing effect is realized by processing the surface of the material into a frosted surface.
In some embodiments, the lighting device can be illuminated in the following ways: 1) annular light reflection type dark field illumination is carried out above the sample, and annular light is distributed on the periphery of the objective lens and is flush with the lower surface of the objective lens;
2) and (3) illuminating the side of the sample, wherein light rays in the light emitting direction of 0 degree of the light source emit to the center of the field of view of the sample within the range of forming an included angle of 10 degrees with the horizontal plane. Preferably, light rays in the light emitting direction of 0 degrees of the light source are incident to the center of the sample view field along the horizontal direction;
3) and annular transmission type dark field illumination is arranged below the sample, and annular light is distributed below the sample and on the periphery of a view field. According to the three illumination modes, the uniform illumination of the surface of the sample is realized, and meanwhile, the phenomenon that exciting light directly enters the lens to form stray light is avoided, so that the imaging effect is influenced.
In addition, the three lighting modes omit the configuration of a dichroic mirror and an excitation filter, so that the space can be effectively reduced, and the compactness of the structure is improved.
In some embodiments, a protective glass 15 is disposed in front of the objective lens 18, which can effectively protect the whole imaging module from contamination by external dust, moisture, etc.
Through implementing the utility model discloses, following beneficial effect has:
1) no additional excitation filtering and dichroic mirrors are required. The absorption spectrum of the fluorescent substance is usually a spectral band, and the spectra of the laser diode and the light emitting diode are narrow, so that the light source with the corresponding wavelength is selected according to the fluorescent substance, and an excitation filter does not need to be added. And because the excitation light path and the detection light path are separately arranged, a dichroic mirror is not needed to separate the excitation light from the fluorescence. The arrangement can effectively reduce the whole volume of the fluorescence microscope and also make the device simpler.
2) The structure is miniature, compact and simple, and is convenient to install and adjust. Because the microscope adopts an approximately symmetrical structure and is combined with the image sensor with high spatial sampling rate, the imaging with large visual field and high resolution can be realized under low magnification, the problem of longer image distance is solved, the conjugate distance is favorably shortened, and the imaging module is miniaturized. In addition, an excitation filter and a dichroic mirror are not needed, so that the occupied space is further reduced while the structure is simplified, and the more compact design is realized.
3) Simple operation and convenient carrying. The fluorescence microscopic imaging module can work only by supplying power through the USB, and the trouble of connecting a mains supply is eliminated. In addition, the mobile phone can be directly connected to display and store images, and the mobile phone is convenient to carry to outdoor use.
In some embodiments, the micro fluorescence microscopy imaging module may further comprise a focus motor 27, optionally the focus motor 27 may be a voice coil motor, stepper motor, ultrasonic motor, memory alloy motor, or the like. The focusing motor 27 can drive the objective lens 18 to realize focusing without influencing the magnification of the system. The focusing motor 27 can also drive the barrel lens 16 or the entire assembly of the objective lens 18, the emission filter 17, and the barrel lens 16 to move to achieve focusing, regardless of the effect on the magnification of the system.
When the focusing motor 27 drives the objective lens 18 or the barrel lens 16 to realize focusing, the object plane position of the micro fluorescence microscopic imaging module is changed. The object plane closest to the micro fluorescence microscopic imaging module is a near-focus object plane, the object plane farthest from the micro fluorescence microscopic imaging module is a far-focus object plane, and the direction far away from the imaging lens module is the positive direction. The objective lens 18 and the cylindrical lens 16 constitute a lens group, the object side of the lens group comprises a limit surface, and when the object side of the lens group is provided with the protective glass 15, the object side surface of the protective glass 15 is the limit surface; when the lens assembly has no protective glass 15 on the object side, the end surface of the housing of the imaging lens module or other mechanical structure matching with the imaging lens module is a limiting surface. The near-focus object plane is positioned in the range of +/-50 mu m of the limiting plane, and the distance between the far-focus object plane and the limiting plane is more than or equal to 220 mu m.
The stroke of the focusing motor 27 is more than or equal to 300 μm and less than or equal to 600 μm, and when the focusing motor 27 drives the objective lens 18 to move, the focusing distance is the same as the stroke of the focusing motor 27. In the focusing range, the minimum distance between the objective lens 18 and the barrel lens 16 is more than or equal to 50 μm. When the protective glass 15 is provided, the minimum distance between the objective lens 18 and the protective glass 15 is more than or equal to 50 μm within the focusing range.
The adoption of the parameters has the following beneficial effects:
1) the near-focus object plane can cover the close-range area of the protective glass 15, can image the close object, and simultaneously, the far-focus object plane can surpass a cover glass commonly used by a microscope, so that the requirements of biomedical imaging are met;
2) the stroke of the focusing motor 27 can effectively cover the dimensional tolerance of each component in the module caused by processing and installation, and the manufacturability of mass production is improved;
3) minimum gaps are reserved among the objective lens 18, the protective glass 15 and the barrel lens 16, so that the reliability of the imaging lens module is improved, and damage caused by mutual collision of components in the module when the focusing motor 27 exceeds a rated stroke is avoided;
4) when realizing above-mentioned beneficial effect, realized the miniaturization of module.
In addition, when the focus motor 27 moves the objective lens 18, the position of the barrel mirror 16 is fixed. The object plane close to focus at this time is the object plane focused by the lens group when the distance between the objective lens 18 and the cylindrical lens 16 is minimum;
when the focus motor 27 moves the barrel mirror 16, the objective lens 18 is fixed in position. The object plane at this time is the object plane focused by the lens group when the distance between the objective lens 18 and the cylindrical lens 16 is minimum.
The following detailed description of the present invention will be made with reference to the accompanying drawings and examples, which are provided for illustration of the present invention and are not intended to limit the scope of the present invention.
In the first embodiment, as shown in fig. 1, the detection optical path is, in order from the object side to the image side, a sample 10, a cover glass 15, an objective lens 18, a barrel mirror 16, an emission filter 17, and an image sensor 28. The objective lens 18 is fixed in the voice coil motor 27, and the barrel mirror 16 is fixed in the barrel mirror holder 12. Wherein the focal length f of the objective lens 1812.6mm, focal length f of the tube mirror 1625.1 mm. An axial distance TTL between an object plane and an image plane of the lens assembly is 10.2mm, and an axial distance TD between an object-side surface and an image-side surface of the lens assembly is 7.2 mm. The emission filter 17 is fixed in the housing 26, and the image sensor 28 is directly integrated on the drive circuit board 11.
The illumination device includes an excitation light source 19 disposed on the periphery of the objective lens 18, and a dodging light guide plate 29 corresponding to the excitation light source 19. The light from the excitation light source 19 passes through the light homogenizing guide 29 to provide uniform illumination of the sample 10. In the present embodiment, the excitation light source 19 includes a plurality of excitation light sources, and is uniformly distributed in an annular shape around the objective lens 18, the dodging light guide plate 29 is an annular light guide plate disposed in the light outgoing direction of the excitation light source 19, and an end surface of the annular light guide plate is substantially flush with an end surface of the objective lens 18 close to the sample 10, so as to form annular light reflection type dark field illumination above the sample. Preferably, the excitation light source 19 is an LED lamp, the LED lamp and the uniform light guide plate 29 are fixed on the housing 26, the LED lamp is at least 2 LED light sources distributed at equal angular intervals along the circumferential direction, and the specific number of the light sources can be set according to parameters such as the illumination uniformity and the illuminance value of the sample surface.
The driving circuit board 11 is provided with a USB interface, which can be directly connected to a computer USB port or connected to a mobile phone through an OTG cable. When the circuit board is electrified, the fluorescence microscope is in a working state. The LED lamp 19 emits excitation light, which passes through the light-homogenizing guide plate 29 and the protective glass 15 and then irradiates the sample 10, thereby forming uniform illumination. The fluorescent substance is excited to generate a fluorescent signal, the fluorescent signal is collected by the objective lens 18 after passing through the protective glass 15, and is finally imaged on the image sensor 28 after sequentially passing through the focusing of the barrel lens 16 and the filtering of the emission optical filter 17. The signal of the image sensor 28 is processed by the driving circuit board 11 and displayed on a computer or a mobile phone through a USB data line, so that an operator can watch the fluorescent image in real time and store the fluorescent image as required.
The voice coil motor 27 is fixed to the housing 26 around the objective lens 18. The driving circuit board 11 drives the voice coil motor 27, and an electrified coil inside the voice coil motor 27 generates an ampere force under the action of a magnetic field, so that the objective lens 18 can move axially, and the automatic focusing function is realized.
In the second embodiment, as shown in fig. 2, the lighting device, the position of the emission filter 17, and the setting of the focusing target are different compared to the first embodiment. The illumination device includes an excitation light source 19 provided on one side of the specimen 10, and light emitted from the excitation light source 19 is irradiated to a target region from the side of the specimen 10. The micro fluorescence microscopic imaging module further comprises a supporting arm 24 arranged on one side of the imaging lens module, and the excitation light source 19 is arranged on the supporting arm 24.
In this embodiment, the micro fluorescence microscopy imaging module further comprises a support base, wherein the support base comprises a support arm 24 and a base 23 fixed at the end of the support arm 24. The excitation light source 19 is a light emitting diode fixed on the support arm 24, and emits light to the target area from the side of the sample, so that the structure is simple and the effect of uniform illumination can be realized. The number of excitation light sources 19 may be distributed along the sample perimeter with support arms 24 connected to the pcb 11 to provide circuit support for the excitation light sources 19 according to actual illumination requirements. The base 23 provides support for the entire fluorescence microscopy imaging module. The emission filter 17 is disposed between the objective lens 18 and the barrel mirror 16, and fixed to the housing 26. The emission filter 17 filters out other unwanted wavelengths of light that pass through the objective lens 18 to avoid aberrations. The voice coil motor 27 is fixed on the housing 26 around the barrel mirror 16, and drives the barrel mirror 16 to move axially, thereby realizing automatic focusing.
In the third embodiment, as shown in fig. 3, the lighting device, the position of the emission filter 17, and the setting of the focusing target are different compared to the first embodiment. The illuminating device is arranged below the sample 10 and comprises at least two first excitation light sources 19 correspondingly arranged below the periphery of the field of view of the objective lens 18, a first dodging light guide plate 29 corresponding to the first excitation light sources 19 and a shielding plate 30 correspondingly arranged at the edge of the field of view of the objective lens 18, and light rays are irradiated to a target area from the lower side of the sample 10.
The illumination device further comprises at least one second excitation light source 19A correspondingly arranged under the field of view of the objective lens 18 and a second dodging light guide plate 29A corresponding to the second excitation light source 19A, and light is irradiated to the target area from right below the sample 10. The shielding plate 30 isolates the first and second excitation light sources 19 and 19A from the first and second uniform light guide plates 29 and 29A, respectively.
In the present embodiment, the excitation light source includes a plurality of excitation light sources, and is uniformly distributed in an annular shape under the periphery of the field of view of the objective lens 18, and the dodging light guide plate is an annular light guide plate disposed in the light emitting direction of the excitation light source.
In this embodiment, the micro fluorescence microscopy imaging module further comprises a support base, wherein the support base comprises a support arm 24 and a base 23 fixed at the end of the support arm 24. The support arm 24 holds the specimen 10 from the side.
In this embodiment, the first excitation light source 19 and the second excitation light source 19A are both LED lamps, the LED lamps are fixed on the base 23 and located at the periphery of the field of view of the objective lens, and the light emitted by the LED lamps passes through the first light homogenizing guide plate 29 and then is irradiated to the target area from the lower side of the sample to form the transmission dark field illumination. The illumination mode is suitable for uniform illumination of the transparent sample, and can also avoid direct incidence of the illumination light into the objective lens. The shielding plate 30 functions to prevent light of the LED lamp 19 from being directly incident to the sample surface from the inside while isolating the first and second uniform light guide plates 29 and 29A. Further, directly below the sample, LED lamps 19A are also distributed, which are independently controllable from the LED lamps 19. When the LED lamp 29A is turned on, light directly irradiates the sample to excite fluorescence after passing through the second uniform light guide plate 29A from right below the sample. The two light source illumination modes can be selected according to different samples or different requirements. The supporting arm 24 is connected to the driving circuit board 11 and provides a circuit support for the ring-shaped light sources, i.e., the first excitation light source 19 and the second excitation light source 19A. The emission filter 17 is disposed in front of the objective lens 18, and can shorten the distance between the objective lens 18 and the tube lens 16, thereby facilitating pupil articulation and better correcting aberration. The voice coil motor 27 is provided at a different position. The voice coil motor 27 is arranged on the periphery of the fixing parts of the objective lens 18, the barrel mirror 16 and the emission light filter 17, and can drive the lens group to move integrally, so that automatic focusing is realized.
In the fourth embodiment, as shown in fig. 4, compared with the second embodiment, the voice coil motor 27 is omitted, and the objective lens 18 is fixed to the objective lens holder 13, so that the whole microscope structure is simpler and more miniaturized. The cover glass 15 is reduced so that it reduces stray light from entering the lens. The light source 22 is a laser diode, has a small divergence angle, and is sufficiently focused by light to be incident on the sample surface.
The present invention has been described in terms of specific embodiments, and it will be understood by those skilled in the art that various changes may be made and equivalents may be substituted without departing from the scope of the invention. In addition, many modifications may be made to adapt a particular situation or material to the teachings of the invention without departing from its scope. Therefore, it is intended that the invention not be limited to the particular embodiment disclosed, but that the invention will include all embodiments falling within the scope of the appended claims.
Claims (12)
1. A micro fluorescence microscopic imaging module is characterized by comprising an imaging lens module and an illuminating device;
the imaging lens module comprises an objective lens (18) and a tube lens (16) which are symmetrically arranged in sequence from an object side to an image side, an image sensor (28) which is arranged on the image side of the tube lens (16) and has high spatial sampling rate, and an emission filter (17) which is arranged in front of the objective lens (18), or between the objective lens (18) and the tube lens (16), or between the tube lens (16) and the image sensor (28);
the illuminating device and the imaging lens module are arranged separately, and uniform illumination on a sample (10) is formed;
an excitation light path of the illuminating device is separated from a detection light path of the imaging lens module; the detection light path comprises a sample (10), an emission filter (17), an objective lens (18), a tube lens (16) and an image sensor (28) from an object side to an image side in sequence; or a sample (10), an objective lens (18), an emission filter (17), a barrel mirror (16) and an image sensor (28); or the sample (10), the objective lens (18), the barrel mirror (16), the emission filter (17) and the image sensor (28).
2. The micro fluorescence microscopy imaging module as claimed in claim 1, wherein the objective lens (18) and the cylindrical lens (16) constitute a lens group, the distance from the object surface to the image surface on the optical axis of the lens group is TTL, and the distance from the object side surface to the image side surface of the lens group on the optical axis is TD, which satisfies the following relationship: TD/TTL is more than or equal to 0.15 and less than or equal to 0.9;
the focal length f of the objective lens (18)1And the focal length f of the barrel mirror (16)2Satisfies the following relationship: f is not less than 0.11/f2≤10；
The optical cylinder length of the lens group satisfies the following relationship: TTL/(f) is not less than 0.21+f2)≤15；
A distance L from an object main plane of the objective (18) to the object planeObjThe following relationship is satisfied: 0.5f1≤LObj≤1.5f1；
The distance L from the image space main plane of the cylindrical mirror (16) to the image surfaceImaSatisfies the following relation: 0.5f2≤LIma≤1.5f2。
3. The micro fluorescence microscopy imaging module according to claim 1, wherein the illumination device comprises an excitation light source (19) disposed at the periphery of the objective lens (18), and a dodging light guide plate (29) corresponding to the excitation light source (19);
the light emitted by the excitation light source (19) passes through the uniform light guide plate (29) to form uniform illumination on the sample (10).
4. The micro fluorescence microscopy imaging module according to claim 3, wherein the excitation light source (19) comprises a plurality of excitation light sources, and is uniformly distributed on the periphery of the objective lens (18) in a ring shape; the uniform light guide plate (29) is an annular light guide plate arranged in the light outgoing direction of the excitation light source (19); and the end face of the annular light guide plate is substantially flush with the end face of the objective lens (18) close to the sample (10).
5. The micro fluorescence microscopy imaging module according to claim 1, wherein the illumination means comprises an excitation light source (19) arranged on one side of the sample (10); the light emitted by the excitation light source (19) is irradiated to a target area from the side of the sample (10).
6. The micro fluorescence microscopy imaging module according to claim 5, further comprising a support arm (24) disposed on one side of the imaging lens module; the excitation light source (19) is mounted on the support arm (24).
7. The micro fluorescence microscopy imaging module of claim 1,
the illuminating device is arranged below the sample (10), and comprises a first excitation light source (19) correspondingly arranged below the periphery of a field of view of the objective lens (18), a first dodging light guide plate (29) corresponding to the first excitation light source (19), and a shielding plate (30) correspondingly arranged at the edge of the field of view of the objective lens (18), wherein light rays are irradiated to a target area from the side lower part of the sample (10).
8. The micro fluorescence microscopy imaging module according to claim 7, wherein the illumination device further comprises a second excitation light source (19A) correspondingly disposed under the field of view of the objective lens (18), and a second dodging light guide plate (29A) corresponding to the second excitation light source (19A), wherein light is irradiated to a target area from right below the sample (10);
the shielding plate (30) isolates the first excitation light source (19) and the second excitation light source (19A), and the first uniform light guide plate (29) and the second uniform light guide plate (29A) respectively.
9. The micro fluorescence microscopy imaging module according to claim 8, further comprising a support base comprising a support arm (24) and a base (23) fixed to a distal end of the support arm (24); the support arm (24) fixes the specimen (10) from the side thereof.
10. The micro fluorescence microscopy imaging module according to any one of claims 3 to 9, wherein the excitation light source (19) is a laser diode or a light emitting diode;
the objective lens (18) and the tube lens (16) are each composed of at least three lenses.
11. The micro fluorescence microscopy imaging module according to any one of claims 1 to 9, further comprising a driving circuit board (11) with a USB interface for driving the illumination means and the image sensor (28); andor or
And a protective glass (15) arranged in front of the objective lens (18).
12. The micro fluorescence microscopy imaging module according to claim 1, further comprising a focusing motor (27) for driving the objective lens (18) or the barrel lens (16) or the whole body composed of the objective lens (18), the emission filter (17) and the barrel lens (16) to move for focusing.
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