CN114136938A - Multifunctional miniature wide-field microscopic imaging device and imaging method thereof - Google Patents
Multifunctional miniature wide-field microscopic imaging device and imaging method thereof Download PDFInfo
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- CN114136938A CN114136938A CN202111358242.4A CN202111358242A CN114136938A CN 114136938 A CN114136938 A CN 114136938A CN 202111358242 A CN202111358242 A CN 202111358242A CN 114136938 A CN114136938 A CN 114136938A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/47—Scattering, i.e. diffuse reflection
- G01N21/4738—Diffuse reflection, e.g. also for testing fluids, fibrous materials
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N2021/6463—Optics
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Abstract
A multifunctional miniature wide-field microscopic imaging device and an imaging method thereof belong to the technical field of microscopic imaging. The upper end of the outer shell is connected with a camera adapter provided with an image sensor circuit board and an image sensor and a light source, a focusing lens, an objective lens, a collimating lens and a light splitting mechanism are arranged in the outer shell, the focusing lens corresponds to the image sensor, the collimating lens is externally provided with the light source, and the light source, the image sensor circuit board and an external data acquisition card are in data connection. The imaging method comprises the following steps: the objective lens and the microscope adapter are connected with the outer shell, and the objective lens is aligned to the sample to be measured; starting a light source to emit light, transmitting the light to a light splitting mechanism and then reflecting an objective lens; the objective lens converges the light to the surface of the measured sample to generate reflected light; the reflected light forms parallel light after passing through the objective lens, and the parallel light reaches the light splitting mechanism and then is transmitted to the focusing objective lens; the focusing objective lens converges the light to the surface of the image sensor for imaging. The invention reduces the volume of the device, and expands the application range and application scene of the micro-imaging device.
Description
Technical Field
The invention relates to a multifunctional miniature wide-field microscopic imaging device and an imaging method thereof, belonging to the technical field of microscopic imaging.
Background
The microscope is an optical instrument formed by one lens or a combination of several lenses, and is a mark for human beings to enter the atomic era. It is mainly used for magnifying the instruments that tiny objects can be seen by naked eyes of people. Microscope spectroscopic microscope and electron microscope: the optical microscope was pioneered in 1590 by jameson in the netherlands. The existing optical microscope can magnify an object by 1600 times, the minimum limit of resolution reaches 1/2 of wavelength, and the length of a mechanical barrel of the domestic microscope is generally 160 mm.
Understanding of technology by modern technological developments has recognized that miniaturized integration is a key advance in facilitating low cost production, often leading to improved performance and unexpected applications. This impact has been manifested in various areas including communications and computer and like technologies.
Optical microscopy is a largely non-integratable technique and remains a large and expensive bench-top instrument.
Most of the existing wide-field microscopes are scientific research instruments and equipment, are large in size and high in price, are generally only suitable for places such as hospitals, research institutes and universities, and cannot be widely used. And advanced microscope operation is relatively complicated and tedious, and personnel who have not been trained by professionals cannot operate the microscope. Meanwhile, the device is not suitable for many fields due to the large volume.
Disclosure of Invention
In order to solve the problems in the background technology, the invention provides a multifunctional miniature wide-field microscopic imaging device and an imaging method thereof.
The invention adopts the following technical scheme: a multifunctional miniature wide-field microscopic imaging device comprises a collimating lens, a light splitting mechanism, an objective lens, a focusing lens, an image sensor, a light source, a camera adapter, an image sensor circuit board and an outer shell; the camera adapter is characterized in that a first optical channel is arranged at the upper end of the outer shell, a second optical channel is arranged at the bottom end of the outer shell, a third optical channel is arranged at the side end of the outer shell, the first optical channel and the second optical channel are coaxially communicated, the third optical channel and the second optical channel are vertically communicated, the upper end of the outer shell is connected with the camera adapter in an adjustable and limiting manner, an image sensor circuit board is arranged at the upper end of the camera adapter, an image sensor is integrated at the lower end of the image sensor circuit board, a focusing lens is arranged at a light outlet of the first optical channel, and the focusing lens is arranged corresponding to the image sensor; an objective lens is arranged in the second optical channel; a collimating mirror is arranged at the outer end part of the third optical channel, a light source is arranged on the outer side of the collimating mirror and connected with the outer shell, and a light splitting mechanism is arranged at the communication part of the third optical channel and the second optical channel and used for reflecting light of the third optical channel into the second optical channel; the light source is in data transmission connection with the image sensor circuit board, and the image sensor circuit board is in data transmission connection with the external data acquisition card.
The invention discloses a still imaging method of a multifunctional miniature wide-field microscopic imaging device, which comprises the following steps:
s1: connecting the objective lens with the outer shell;
s2: aligning the focal plane of the objective lens to the sample to be measured;
s3: turning on a light source;
s4: light rays emitted by the light source form parallel light beams after passing through the collimating lens for transmission;
s5: after the parallel light beams reach the light splitting mechanism, the parallel light beams are reflected to change the propagation direction to the objective lens;
s6: the objective lens converges the parallel light beams to the surface of the measured sample;
s7: the measured sample generates reflected light;
s8: the reflected light is modulated by the objective lens to form parallel light, and the parallel light reaches the light splitting mechanism and then is transmitted to the focusing objective lens;
s9: the focusing objective lens converges the parallel light to the surface of the image sensor for imaging.
The invention discloses a living animal imaging method of a multifunctional miniature wide-field microscopic imaging device, which comprises the following steps:
s1: connecting the objective lens with the outer shell;
s2: mounting a first microscope adapter at the lower end of the outer shell;
s3: aligning the focal plane of the objective lens to the sample to be measured;
s4: turning on a light source;
s5: light rays emitted by the light source form parallel light beams after passing through the collimating lens for transmission;
s6: after the parallel light beams reach the light splitting mechanism, the parallel light beams are reflected to change the propagation direction to the objective lens;
s7: the objective lens converges the parallel light beams to the surface of the measured sample;
s8: the measured sample generates reflected light;
s9: the reflected light is modulated by the objective lens to form parallel light, and the parallel light reaches the light splitting mechanism and then is transmitted to the focusing objective lens;
s10: the focusing objective lens converges the parallel light to the surface of the image sensor for imaging.
The invention discloses a laboratory sample imaging method of a multifunctional miniature wide-field microscopic imaging device, which comprises the following steps:
s1: connecting the objective lens with the outer shell;
s2: connecting a microscope adapter II, an optical connecting rod I, a connecting rod adapter, an optical connecting rod II and an XYZ displacement table at one time;
s3: mounting a first microscope adapter at the lower end of the outer shell, and mounting a second microscope adapter at the lower end of the first microscope adapter;
or directly mounting the microscope adapter II at the lower end of the outer shell;
s4: aligning the focal plane of the objective lens to the sample to be measured;
s5: turning on a light source;
s6: light rays emitted by the light source form parallel light beams after passing through the collimating lens for transmission;
s7: after the parallel light beams reach the light splitting mechanism, the parallel light beams are reflected to change the propagation direction to the objective lens;
s8: the objective lens converges the parallel light beams to the surface of the measured sample;
s9: the measured sample generates reflected light;
s10: the reflected light is modulated by the objective lens to form parallel light, and the parallel light reaches the light splitting mechanism and then is transmitted to the focusing objective lens;
s11: the focusing objective lens converges the parallel light to the surface of the image sensor for imaging.
Compared with the prior art, the invention has the beneficial effects that:
1. on the premise that core indexes such as a view field, resolution and the like are not weaker than those of a traditional desk microscope, the volume of the desk microscope is greatly reduced, and the advantages of microscopic imaging can be revealed in many fields which cannot be realized originally;
2. by arranging the light splitting mechanism and the lens with the gradient refractive index of 8 degrees, the micro-imaging device can perform imaging detection on a fluorescent sample and non-fluorescent samples (such as undyed animal and plant sample slices, industrial chips and other samples), and the application range and application scene of the micro-imaging device are expanded.
Drawings
FIG. 1 is a schematic structural view of the present invention;
FIG. 2 is a cross-sectional view of FIG. 1;
FIG. 3 is an exploded view of FIG. 1;
FIG. 4 is a schematic view of the mounting of the objective lens;
FIG. 5 is a schematic view of the structure of the objective lens;
FIG. 6 is a schematic view of the mounting of the second microscope adapter to the outer housing;
FIG. 7 is a schematic view of the installation of the first microscope adapter, the second microscope adapter and the outer housing;
FIG. 8 is a use state reference diagram of the present invention;
FIG. 9 is an image of an onion section specimen taken in accordance with the present invention;
FIG. 10 is an image of a kidney section specimen in accordance with the present invention;
FIG. 11 is an image of a wafer structure according to the present invention;
FIG. 12 is an image of the present invention taken with a standard resolution plate.
Detailed Description
The technical solutions in the present invention will be described clearly and completely with reference to the accompanying drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the invention, rather than all embodiments, and all other embodiments obtained by those skilled in the art without any creative work based on the embodiments of the present invention belong to the protection scope of the present invention.
A multifunctional miniature wide-field microscopic imaging device comprises a collimating lens 1, a light splitting mechanism 3, an objective lens 4, a focusing lens 6, an image sensor 7, a light source 8, a camera adapter 14, an image sensor circuit board 17 and an outer shell 22; the outer shell 22 is a carrier of all optical elements and circuit elements of the present invention, and is made of black resin, or any hard metal or nonmetal material, the upper end of the outer shell 22 is provided with a first optical channel, the bottom end of the outer shell 22 is provided with a second optical channel, and the side end of the outer shell 22 is provided with a third optical channel, the first optical channel and the second optical channel are coaxially communicated to form a reflective optical path; the third light channel is vertically communicated with the second light channel to form an illumination light path; the illumination light path is as follows according to the light propagation direction in sequence: the light source 8, the collimating mirror 1, the light splitting mechanism 3 and the objective 4, the transmission direction of the reflected light path illumination light is as follows in sequence: the objective lens 4, the light splitting mechanism 3, the focusing lens 6 and the image sensor 7; the upper end of the outer shell 22 is connected with the camera adaptor 14 in an adjustable limit manner (the camera adaptor 14 is responsible for connecting the image sensor circuit board 17 with the outer shell 22. in the invention, the lower half part of the camera adaptor 14 is a male sliding groove which is in sliding clearance fit with a female sliding groove of the upper half part of the outer shell 22, the adjustable limit fixing connection is carried out through a first jackscrew 18 and a second jackscrew 19 which are arranged on the lateral side of the male sliding groove of the camera adaptor 14, the outer shell 22 and the side wall of the camera adaptor 14 are correspondingly provided with two threaded holes M1.4, but not limited to the range, the first jackscrew 18 and the second jackscrew 19 are arranged in the corresponding threaded holes), the upper end of the camera adaptor 14 is provided with the image sensor circuit board 17 (the upper half part of the camera adaptor 14 is fixedly connected with the image sensor circuit board 17 through a fourth fastening screw 15, a fifth fastening screw 16 and a sixth fastening screw 23, keeping the volume to a minimum. Since the objective lens 4 is fixed, the camera adaptor 14 needs to move up and down to find the focusing plane, and besides the sliding adjustment mode, the adjustment mode can also be in the form of screw rotation, sawtooth groove and the like. The camera adaptor 14 is made of resin, and may be made of any hard metal or nonmetal), the image sensor 7 is integrated at the lower end of the image sensor circuit board 17, the image sensor circuit board 17 is used for controlling parameters such as exposure gain of the image sensor 7, the light outlet of the first optical channel is provided with the focusing lens 6, and the focusing lens 6 is arranged corresponding to the image sensor 7; an objective lens 4 is arranged in the second optical channel; a collimating mirror 1 is arranged at the outer end part of the third optical channel, a light source 8 is arranged on the outer side of the collimating mirror 1 through a third fastening screw 20, the light source 8 is connected with an outer shell 22, a light splitting mechanism 3 is arranged at the communication part of the third optical channel and the second optical channel, and the light splitting mechanism 3 is used for reflecting the light of the third optical channel into the second optical channel;
the objective lens 4 is an infinite objective lens, the gradient index lens is selected, more specifically, the gradient index lens with an angle of 8 degrees of 0.23 pitch is selected, and the inclined plane needs to be arranged close to one side of the light splitting mechanism 3. The gradient index lens with two end faces perpendicular to the optical axis is used, imaging cannot be performed, the illumination light is modulated and reflected by the light splitting mechanism, then vertically enters the first end face of the objective lens 4, and then is focused to the measured sample through the second end face of the objective lens 4 to generate reflected light. The reflected light passes through the objective lens 4, the beam splitting mechanism 3 and the focusing lens 6 and then is converged to the image sensor 7 for imaging.
Multiple experiments prove that the first end face of the gradient refractive index lens can generate reflected light, the intensity of the reflected light is similar to that of signal light, serious noise is generated, and clear imaging cannot be achieved. The use of the 8 DEG gradient index lens can ensure that the first section of surface reflected light does not directly enter the image sensor 7, and the reflected signal light which enters the image sensor 7 and is generated only by the sample can be clearly imaged.
The objective lens 4 may be a plano-convex lens, a cemented lens, a spherical lens, or the like, which has a larger aberration than a gradient index lens, or may be a metamaterial lens attached to a glass substrate. The diameter d2 of the objective 4 is dimensioned such that: d2 is more than 25.4mm and more than or equal to 0.1mm, and the working distance of the focal length phi meets the following requirements: the focal length selection can affect the indexes of imaging field of view, resolution, magnification and the like when phi is more than 30mm and is more than or equal to 0 mm.
The light source 8 is an LED light source or an optical fiber, the LED light source is a white light source, the wavelength is 400-750nm, the LED light source can also be a non-visible near-infrared light source, and can also be a narrow-band light source with any bandwidth, and the light source selection can influence the imaging resolution according to the airy disc formula of 0.61 lambda/NA; the light source may be a laser, a halogen lamp, or other types of light sources. The LED patches are packaged on the light source 8, the packages can be 0402, 0603 and the like, the thickness is 0.8mm but not limited to the thickness, and the LED patches are manufactured into a heat dissipation hole process and have any color.
The collimating lens 1 can be any optical element or non-optical element for collimating light, such as a hemispherical lens, a spherical lens, a drum lens, a gradient index lens, etc., the lens diameter d1 can be 25.4mm > d1 ≧ 0.1mm, 25.4mm is a common size for conventional optical elements, and the size is not limited to miniaturization or miniaturization equipment. The non-optical element may be a metamaterial lens that may be attached to a smaller and thinner glass substrate, making the overall size lighter and smaller.
The focusing lens 6 is an achromatic cemented lens, in the invention, the light source wavelength is 400-750nm, and the imaging quality can be improved by using the achromatic lens. The focusing lens 6 may be a plano-convex lens, a spherical lens, or the like. The diameter d3 of the focusing lens 6 has dimensions such that: 25.4mm > d3 is more than or equal to 1mm, and the common focal length is 5mm, 7mm, 10mm, 15mm, 20mm and the like, but is not limited in the range.
The image sensor 7 is a color or black and white image sensor. The pixel number is 752 multiplied by 480(360960 pixels), the pixel size is 6 mu m, the maximum pixel sampling frame rate can reach 60fps, the shutter efficiency is more than 99 percent, and the double-line serial interface, the Automatic Exposure Control (AEC) and the Automatic Gain Control (AGC), the AEC/AGC with variable regions and variable weights meet the imaging requirements under most conditions.
The light splitting mechanism 3 is a spectroscope or a light splitting film, and when the light splitting mechanism 3 is the spectroscope, the spectroscope is 50: in the 50 spectroscopes, 50% of light energy is lost when incident light passes through the spectroscope mechanism 3 once, and in the present invention, the light energy reaching the image sensor 7 is lost by about 75% in total, regardless of the lost energy of the sample reflected light. When the light-splitting mechanism 3 is a light-splitting film, the weight and volume of the micro-imaging device are further reduced.
The light source 8 is connected with the image sensor circuit board 17 through a single-core lead 39 in a data transmission mode, and the on-off and the brightness of the LED light source circuit board 8 can be controlled. The image sensor circuit board 17 is connected with an external data acquisition card through a flexible shielding wire 38 for data transmission, power supply and data transmission are carried out, and the torque of the miniature wide-field microscopic imaging device can be increased by using the flexible wire to prevent the miniature wide-field microscopic imaging device from being damaged;
the vertical plane of the detection light path is an X-Y plane, the parallel is a Z axis, the size in the XY direction is about 1cm multiplied by 1cm, the size in the Z direction is about 2cm, and the volume of the miniature wide-field microscopic imaging device is about 2cm3The weight was about 2 g. Due to different design schemes, the size in XYZ direction can be expanded to 0.1cm to 20cm, and the volume can be 0.1cm3To 100cm3The weight can be 0.1g to 1000g, the sizes are not limited to the range, and the overlarge value is not a micro microscopic imaging device in a macroscopic sense but a traditional large-volume table-type microscope.
An objective lens adapter 11 is arranged between the objective lens 4 and the second optical channel, the objective lens adapter 11 is responsible for connecting the objective lens 4 and the outer shell 22, the diameter of the objective lens 4 can be designed to be consistent with the diameter of the second optical channel, but the wider application of the objective lens is limited, so the diameter of the second optical channel is slightly larger than the diameter of the objective lens 4, and the objective lens adapter 11 needs to be designed to fixedly connect the objective lens 4 and the outer shell 22. The objective lens adapter 11 is tightly fitted on the outside of the objective lens 4, and the objective lens adapter 11 is tightly inserted into the second optical channel. The objective adapter 11 is made of resin and manufactured by using a 3D printing technology, and may be made of metal or nonmetal such as copper, iron, and aluminum alloy.
The outer wall of the objective lens adapter piece 11 is provided with a limiting protrusion along the circumferential direction, the limiting protrusion is arranged on the outer side of the outer shell 22, the limiting protrusion limits the position of the objective lens adapter piece 11 penetrating into the second optical channel, and collision between the objective lens 4 and the light splitting mechanism 3 is prevented.
The outer side of the outer shell 22 is detachably and fixedly connected with the lens protection cover 25 through a first fastening screw 13 and a second fastening screw 21 (the size of the fastening screw is M1 multiplied by 3, but not limited to the range), a magnet, a mortise and tenon structure, a groove structure, glue fixation and the like.
The lens protective cover 25 can protect the internal optical elements from being polluted, and can also enable the light path transmission route to be in a completely closed state, so that the influence of stray light is reduced, and the imaging quality is improved. The lens protection cover 25 may be made of resin, and may be made of any hard metal or nonmetal.
The lower end of the outer shell 22 is provided with a first microscope adapter 12 and/or a second microscope adapter 31, and when the lower end of the outer shell 22 is provided with the first microscope adapter 12 and the second microscope adapter 31, the second microscope adapter 31 is arranged at the lower end of the first microscope adapter 12.
The lower end of the outer shell 22 is detachably and fixedly connected with the microscope adaptor I12.
The first microscope adapter 12 and the outer shell 22 can also be fixed by using structures such as threads, clamping grooves, mortise and tenon joints and the like. All the magnets of the present invention have a diameter of 1mm and a thickness of 1mm, but are not limited thereto.
The jackscrew four 26 fixes the objective adapter 11 and the outer housing 22. The back surface of the microscope adapter piece I12 is provided with a U-shaped groove to avoid the jackscrew II 26, so that the microscope adapter piece I12 and the objective lens adapter piece 11 do not conflict when being used simultaneously.
The microscope adaptor two 31 is arranged at one end of the optical extension rod one 32, the other end of the optical extension rod one 32 is arranged on the extension rod adaptor 33, the extension rod adaptor 33 is arranged on the XYZ displacement table 35, and the optical extension rod two 34 is arranged on the extension rod adaptor 33.
The present invention can constitute a bench microscope with an extremely simple structure as shown in fig. 6, and functions as the conventional bench microscope. The optical extension rod one 32, the extension rod adaptor 33, the optical extension rod two 34 and the XYZ displacement table 35 used in fig. 6 are all the most commonly used optical-mechanical devices and displacement platforms in a laboratory, fig. 6 only shows an assembly manner, and in practical applications, there are various alternatives, such as the length of the optical extension rod, the connection manner, and the like, and the displacement table can be a single-axis displacement table, a double-axis displacement table, a three-axis displacement table, a four-axis displacement table, a five-axis displacement table, a six-axis displacement table, and the like.
The microscope adapter piece II 31 has two types which can be selected, the internal threads corresponding to the side surface of the microscope adapter piece II 31 are M3 and M4, and the two types are common adapter types of laboratory optical connecting rods. The bottom end of the microscope adaptor second 31 can be connected with the microscope adaptor first 12 through three magnets at corresponding positions, and can also be connected with the outer shell 22, and using methods are different according to different requirements. The material of the second microscope adaptor 31 is resin, and may be any hard metal or nonmetal material.
A method of still imaging for a multi-functional miniature wide-field microscopic imaging device, said method comprising the steps of:
s1: connecting the objective 4 to the outer housing 22 via the objective adapter 11;
s2: the invention is held by hand, the focal plane of the objective lens 4 is aligned with the sample to be measured;
s3: turning on the light source 8;
s4: light rays emitted by the light source 8 form parallel light beams after passing through the collimating lens 1 for transmission;
s5: after the parallel light beams reach the light splitting mechanism 3, the parallel light beams are reflected to change the propagation direction to the objective lens 4;
s6: the objective lens 4 converges the parallel light beams to the surface of the measured sample;
s7: the measured sample generates reflected light;
s8: the reflected light is modulated by the objective lens 4 to form parallel light, and the parallel light reaches the light splitting mechanism 3 and then is transmitted to the focusing objective lens 6;
s9: the focusing objective 6 focuses the parallel light to the surface of the image sensor 7 for imaging.
A method for imaging a living animal with a multifunctional miniature wide-field microscopic imaging device, the method comprising the steps of:
s1: connecting the objective 4 to the outer housing 22 via the objective adapter 11;
s2: mounting the microscope adaptor one 12 at the lower end of the outer shell 22;
s3: aligning the focal plane of the objective lens 4 to the sample to be measured;
s4: turning on the light source 8;
s5: light rays emitted by the light source 8 form parallel light beams after passing through the collimating lens 1 for transmission;
s6: after the parallel light beams reach the light splitting mechanism 3, the parallel light beams are reflected to change the propagation direction to the objective lens 4;
s7: the objective lens 4 converges the parallel light beams to the surface of the measured sample;
s8: the measured sample generates reflected light;
s9: the reflected light is modulated by the objective lens 4 to form parallel light, and the parallel light reaches the light splitting mechanism 3 and then is transmitted to the focusing objective lens 6;
s10: the focusing objective 6 focuses the parallel light to the surface of the image sensor 7 for imaging.
A method for laboratory sample imaging with a multifunctional miniature wide-field microscopy imaging device, said method comprising the steps of:
s1: connecting the objective 4 to the outer housing 22 via the objective adapter 11;
s2: connecting a microscope adaptor II 31, an optical connecting rod I32, a connecting rod adaptor 33, an optical connecting rod II 34 and an XYZ displacement table 35 at one time;
s3: installing the microscope adapter piece I12 at the lower end of the outer shell 22, and then installing the microscope adapter piece II 31 at the lower end of the microscope adapter piece I12;
or directly mounting the second microscope adapter 31 at the lower end of the outer shell 22;
s4: aligning the focal plane of the objective lens 4 to the sample to be measured;
s5: turning on the light source 8;
s6: light rays emitted by the light source 8 form parallel light beams after passing through the collimating lens 1 for transmission;
s7: after the parallel light beams reach the light splitting mechanism 3, the parallel light beams are reflected to change the propagation direction to the objective lens 4;
s8: the objective lens 4 converges the parallel light beams to the surface of the measured sample;
s9: the measured sample generates reflected light;
s10: the reflected light is modulated by the objective lens 4 to form parallel light, and the parallel light reaches the light splitting mechanism 3 and then is transmitted to the focusing objective lens 6;
s11: the focusing objective 6 focuses the parallel light to the surface of the image sensor 7 for imaging.
The object provided by the invention is really manufactured according to the invention, can be used, and does not have false enlarged protection range. The eighth group of elements in the fifth row of the standard resolution plate can be resolved in fig. 12, the resolution is about 1 μm, the diameter of the field of view is about 2mm, but the resolution and the field of view are not limited to the sub-resolution and the field of view, and the resolution and the field of view of the optical system are mutually restricted and can be higher or lower, fig. 9 and 10 show the imaging effect of the biological sample, fig. 11 shows the imaging effect of the industrial sample, and the structures of the samples in fig. 9, 10 and 11 are all in the micron level.
The prior art adopts a fluorescence detection method, a spectroscope is a dichroic mirror and only modulates specified wavelength, a detected sample can excite reflected light larger than the wavelength of illumination light, and the light splitting mechanism of the invention can not differentially modulate full-wavelength light, and the detected sample only generates reflected light with the same wavelength.
In the prior art, the objective lens is generally a gradient index lens, and two end faces are planes perpendicular to an optical axis, while the end face of the gradient index lens is perpendicular to the optical axis, and the other end face of the gradient index lens forms an included angle of 8 degrees with the optical axis.
The invention manufactures a miniature wide-field microscopic imaging device by elaborately designing an integral structure, customizing or independently designing appropriate optical elements such as an objective lens, a tube lens and the like, a miniature-sized CMOS sensor and a data acquisition card. On the premise that core indexes such as a view field and resolution are not weaker than that of a traditional table microscope, the size of the equipment is greatly reduced, and the advantages of microscopic imaging, such as detection of slits or the inside of a pipeline of large-scale industrial equipment, can be revealed in a plurality of fields which cannot be realized originally. In addition, the multifunctional multiplexing of a handheld microscope, a suspended microscope and a bench microscope can be realized through the microscope adaptor designed by the user. Allowing mass production and maintaining high stability comparable to conventional microscopes.
The micro-microscopy technique is based on micro-optics and semi-conductor, both of which are easy to manufacture in large quantities and at low cost, and our device has significant advantages over high resolution fiber optic microscopes in terms of optical sensitivity, field of view, resolution, mechanical flexibility, cost and portability.
It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein. Any reference sign in a claim should not be construed as limiting the claim concerned.
Furthermore, it should be understood that although the present description refers to embodiments, not every embodiment may contain only a single embodiment, and such description is for clarity only, and those skilled in the art should integrate the description, and the embodiments may be combined as appropriate to form other embodiments understood by those skilled in the art.
Claims (9)
1. A multifunctional miniature wide-field microscopic imaging device is characterized in that: the device comprises a collimating lens (1), a light splitting mechanism (3), an objective lens (4), a focusing lens (6), an image sensor (7), a light source (8), a camera adapter (14), an image sensor circuit board (17) and an outer shell (22); the upper end of the outer shell (22) is provided with a first optical channel, the bottom end of the outer shell (22) is provided with a second optical channel, a third optical channel is arranged at the side end of the outer shell (22), the first optical channel and the second optical channel are coaxially communicated, the third optical channel and the second optical channel are vertically communicated, the upper end of the outer shell (22) is connected with the camera adaptor (14) in an adjustable and limiting manner, the upper end of the camera adaptor (14) is provided with an image sensor circuit board (17), the lower end of the image sensor circuit board (17) is integrated with an image sensor (7), a light outlet of the first optical channel is provided with a focusing lens (6), and the focusing lens (6) and the image sensor (7) are correspondingly arranged; an objective lens (4) is arranged in the second optical channel; a collimating mirror (1) is arranged at the outer end part of the third optical channel, a light source (8) is arranged on the outer side of the collimating mirror (1), the light source (8) is connected with an outer shell (22), a light splitting mechanism (3) is arranged at the communication part of the third optical channel and the second optical channel, and the light splitting mechanism (3) is used for reflecting the light of the third optical channel into the second optical channel; the light source (8) is in data transmission connection with the image sensor circuit board (17), and the image sensor circuit board (17) is in data transmission connection with the external data acquisition card.
2. The multifunctional miniature wide-field microscopic imaging device according to claim 1, wherein: an objective lens adapter (11) is arranged between the objective lens (4) and the second optical channel, the objective lens adapter (11) is tightly sleeved on the outer side of the objective lens (4), and the objective lens adapter (11) is tightly inserted into the second optical channel.
3. The multifunctional miniature wide-field microscopic imaging device according to claim 2, wherein: the outer wall of objective lens adaptor (11) is equipped with spacing arch along its circumference, spacing arch sets up the outside in shell (22).
4. The multifunctional miniature wide-field microscopic imaging device according to claim 1 or 3, wherein: the outer side of the outer shell (22) is detachably and fixedly connected with the lens protection cover (25).
5. The multifunctional miniature wide-field microscopic imaging device according to claim 4, wherein: the microscope adapter is characterized in that a first microscope adapter (12) and/or a second microscope adapter (31) are/is arranged at the lower end of the outer shell (22), and when the first microscope adapter (12) and the second microscope adapter (31) are arranged at the lower end of the outer shell (22), the second microscope adapter (31) is arranged at the lower end of the first microscope adapter (12).
6. The multifunctional miniature wide-field microscopic imaging device according to claim 5, wherein: the microscope adaptor II (31) is arranged at one end of the optical extension rod I (32), the other end of the optical extension rod I (32) is arranged on the extension rod adaptor (33), the extension rod adaptor (33) is arranged on the XYZ displacement table (35), and the optical extension rod II (34) is arranged on the extension rod adaptor (33).
7. A still image forming method of the multi-functional miniature wide-field microscopic imaging device according to any one of claims 1-6, characterized in that: the method comprises the following steps:
s1: connecting the objective lens (4) with the outer housing (22);
s2: aligning the focal plane of the objective lens (4) to the sample to be measured;
s3: turning on a light source (8);
s4: light rays emitted by the light source (8) form parallel light beams after passing through the collimating lens (1) for transmission;
s5: after the parallel light beams reach the light splitting mechanism (3), the parallel light beams are reflected to change the propagation direction to the objective lens (4);
s6: the objective lens (4) converges the parallel light beams to the surface of the sample to be measured;
s7: the measured sample generates reflected light;
s8: the reflected light is modulated by the objective lens (4) to form parallel light, and the parallel light reaches the light splitting mechanism (3) and then is transmitted to the focusing objective lens (6);
s9: the focusing objective lens (6) converges the parallel light to the surface of the image sensor (7) for imaging.
8. A method for imaging a living animal by using the multi-functional miniature wide-field microscopic imaging device according to any one of claims 5-6, wherein the method comprises the following steps: the method comprises the following steps:
s1: connecting the objective lens (4) with the outer housing (22);
s2: mounting a microscope adaptor I (12) at the lower end of the outer shell (22);
s3: aligning the focal plane of the objective lens (4) to the sample to be measured;
s4: turning on a light source (8);
s5: light rays emitted by the light source (8) form parallel light beams after passing through the collimating lens (1) for transmission;
s6: after the parallel light beams reach the light splitting mechanism (3), the parallel light beams are reflected to change the propagation direction to the objective lens (4);
s7: the objective lens (4) converges the parallel light beams to the surface of the sample to be measured;
s8: the measured sample generates reflected light;
s9: the reflected light is modulated by the objective lens (4) to form parallel light, and the parallel light reaches the light splitting mechanism (3) and then is transmitted to the focusing objective lens (6);
s10: the focusing objective lens (6) converges the parallel light to the surface of the image sensor (7) for imaging.
9. A laboratory sample imaging method of the multifunctional miniature wide-field microscopic imaging device according to any one of claims 5 to 6, characterized in that: the method comprises the following steps:
s1: connecting the objective lens (4) with the outer housing (22);
s2: connecting a microscope adaptor II (31), an optical extension rod I (32), an extension rod adaptor (33), an optical extension rod II (34) and an XYZ displacement table (35) at one time;
s3: installing a microscope adapter piece I (12) at the lower end of the outer shell (22), and then installing a microscope adapter piece II (31) at the lower end of the microscope adapter piece I (12);
or directly installing a second microscope adapter (31) at the lower end of the outer shell (22);
s4: aligning the focal plane of the objective lens (4) to the sample to be measured;
s5: turning on a light source (8);
s6: light rays emitted by the light source (8) form parallel light beams after passing through the collimating lens (1) for transmission;
s7: after the parallel light beams reach the light splitting mechanism (3), the parallel light beams are reflected to change the propagation direction to the objective lens (4);
s8: the objective lens (4) converges the parallel light beams to the surface of the sample to be measured;
s9: the measured sample generates reflected light;
s10: the reflected light is modulated by the objective lens (4) to form parallel light, and the parallel light reaches the light splitting mechanism (3) and then is transmitted to the focusing objective lens (6);
s11: the focusing objective lens (6) converges the parallel light to the surface of the image sensor (7) for imaging.
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| CN115061270A (en) * | 2022-05-30 | 2022-09-16 | 中国人民解放军国防科技大学 | Tilt-mode telescopic microscope combined imaging method |
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