CN218099754U - Line scanning type fluorescence microscope using multi-wavelength laser light source - Google Patents

Abstract

The utility model discloses an use line scanning formula fluorescence microscope of multi-wavelength laser light source, including beam combination optical group, powell prism, cylindrical mirror, exciting light filter wheel, beam splitter, microobjective, transmission light filter wheel, tube lens and TDI linear array camera, beam combination optical group, powell prism, cylindrical mirror, exciting light filter wheel, beam splitter set up along light incidence direction in proper order, and microobjective sets up in beam splitter one side, and the opposite side of beam splitter is provided with tube lens and TDI linear array camera. The utility model discloses in, adopt the excitation light source that the multi-wavelength laser was restrainted, compare in the LED light source that current fluorescence microscope adopted, luminance is higher, and the volume is littleer, and the cost is lower, simultaneously, has carried out the plastic to excitation light source, has obtained the better linear facula of homogeneity, matches TDI linear array camera's clear aperture more, has reduced the optical power loss that causes when using circular facula to incide.

Description

Line scanning type fluorescence microscope using multi-wavelength laser light source

Technical Field

The utility model belongs to the technical field of fluorescence microscope, concretely relates to use line scanning formula fluorescence microscope of multi-wavelength laser light source.

Background

Some substances can emit light with a wavelength longer than that of the excitation light under the irradiation of light with a specific wavelength range, namely fluorescence, and the fluorescence microscope utilizes the optical principle. The fluorescence microscope is mainly used for researching absorption and transportation of substances in cells, distribution and positioning of chemical substances and the like. The fluorescence microscope mainly comprises an excitation light source, an excitation optical filter, a beam splitter, an emission optical filter, a microscope objective and a camera. Most of excitation light sources adopted by the existing fluorescence microscope are LED light sources which can emit light with a plurality of different wavelengths, and in order to meet the requirement of high brightness, the LED light sources need to adopt arrays, and the LED light sources are generally large in size. With the development of laser, some high-end fluorescence microscopes use laser as an excitation light source, and compared with an LED light source, the laser has higher light energy density, lower cost, and smaller volume under the same light power.

In a fluorescence microscope using a TDI line camera, if a traditional LED light source is used, a great optical power loss is caused because the clear aperture of the TDI line camera is linear, such as rectangular, and the light spot emitted by the LED light source is a circular light spot, when the TDI line camera is covered by the whole circular light spot, the light beam is lost completely except for the part with the rectangular light aperture, which can enter the TDI line camera, so that the light beam loss is reduced or even avoided, and the light spot emitted by the LED light source needs to be shaped into the linear light spot matched with the TDI line camera through a light beam shaping structure. However, the LED light source generally has a large volume, a low light energy density compared to laser light, and a large divergence angle, so that the difficulty and cost of shaping the LED light source are large, and the illuminance after shaping is uneven. In addition, if the LED light source is required to achieve the same brightness as the laser, a plurality of LED light sources need to be arrayed to emit light simultaneously, which may increase the volume and cost.

It should be noted that even though the fluorescence microscope using the TDI line camera uses a laser light source, if the beam shaping is not performed, the problem of light loss due to incomplete adaptation to the linear light spot of the TDI line camera also occurs. However, no such beam shaping structure is found in the fluorescence microscope using the TDI line camera in the market at present, and a market blank exists.

Under the premise of shaping the light beam, in order to ensure the uniformity of the illumination of the light beam and reduce the volume as much as possible, a laser light source is more suitable. Therefore, the utility model provides a line scanning type fluorescence microscope using multi-wavelength laser light source.

SUMMERY OF THE UTILITY MODEL

In order to solve the technical problem existing in the prior art, an object of the present invention is to provide a line scanning fluorescence microscope using a multi-wavelength laser light source.

In order to realize the purpose, the technical effect is achieved, the utility model adopts the technical scheme that:

a line scanning type fluorescence microscope using a multi-wavelength laser light source comprises a beam combination light group, a Powell prism, a cylindrical mirror, an exciting light filter wheel, a beam splitter, a microscope objective, an emitted light filter wheel, a tube lens and a TDI linear array camera, wherein the beam combination light group, the Powell prism, the cylindrical mirror, the exciting light filter wheel and the beam splitter are sequentially arranged along a light incidence direction, the microscope objective is arranged on one side of the beam splitter, and the tube lens and the TDI linear array camera are arranged on the other side of the beam splitter.

Furthermore, beam combination optical group includes laser group, collimating lens group and dichroic mirror group that set gradually along light incidence direction, laser group includes the laser instrument of a plurality of wavelength diverse, collimating lens group includes a plurality of collimating lens, dichroic mirror group includes a plurality of dichroic mirror, and laser instrument, collimating lens and dichroic mirror's quantity is the same, and every laser instrument all corresponds with a collimating lens and a dichroic mirror.

Further, the laser group includes four lasers with different wavelengths, which are sequentially marked as: the device comprises a laser I, a laser II, a laser III and a laser IV; the collimating lens group comprises four collimating lenses which are sequentially arranged from left to right; dichroic mirror group includes four dichroic mirrors, records as in proper order from left to right: a dichroic mirror I, a dichroic mirror II, a dichroic mirror III and a dichroic mirror IV; the laser device I, the leftmost collimating lens and the dichroic mirror I are in one-to-one correspondence and adaptation in position, the laser device II, the middle left collimating lens and the middle right collimating lens are in one-to-one correspondence and adaptation in position, the laser device III, the middle right collimating lens and the dichroic mirror III are in one-to-one correspondence and adaptation in position, and the laser device IV, the rightmost collimating lens and the dichroic mirror IV are in one-to-one correspondence and adaptation in position.

Further, the wavelength range of the laser is 330 nm-550 nm.

Furthermore, the collimating lens is a spherical lens, an aspheric lens, a cylindrical lens or a spherical lens, and is coated with an antireflection film.

Furthermore, the diameter of an emergent light spot of the dichroic mirror group is 0.8-3 mm, and the diameter of a light spot incident on the Bawell prism is 0.8-3 mm.

Furthermore, an antireflection film is plated on the surface of the Bawell prism, the fan angle of the Bawell prism is 10-60 degrees, and light spots emitted by the Bawell prism are linear light spots.

Furthermore, the line view field of the microscope objective is 10-30 mm, the numerical aperture NA is 0.1-1.25, and the magnification is 4X-100X.

Furthermore, the focal length of the tube lens is 180 mm-200 mm, and the linear field of view is 10-30 mm.

Furthermore, the excitation light filter wheel and the emission light filter wheel have the same structure and respectively comprise a panel, a rotating motor M and a filter wheel, four filters with the same diameter are uniformly distributed on the panel, and the filter wheel is connected with the rotating motor M and controlled by the rotating motor M.

Compared with the prior art, the beneficial effects of the utility model are that:

the utility model discloses an use line scanning formula fluorescence microscope of multi-wavelength laser light source, compare in the LED light source that current fluorescence microscope adopted, the utility model discloses an excitation light source that different wavelength lasers restrainted, its luminance is higher, and the monochromaticity of laser is better, and a laser instrument corresponds a wavelength, can adapt to the requirement that fluorescence microscope needs different wavelength light-emitting, even combine a plurality of lasers of different wavelength, the volume of the laser group after the beam combination still is less than the volume of LED light source, satisfies the volume littleer, the lower demand of cost; meanwhile, in order to match the clear aperture of the TDI linear array camera, the Bawell prism, the cylindrical mirror, the exciting light filter wheel, the beam splitter, the emitted light filter wheel and the like are adopted for shaping the exciting light source, so that linear light spots with better uniformity are obtained, the optical power loss caused by the incidence of circular light spots is reduced, the shaping difficulty is lower, the cost is lower, and the method is more suitable for industrial popularization and use.

Drawings

Fig. 1 is a schematic structural view of the present invention;

fig. 2 is a schematic structural diagram of the powell prism of the present invention;

fig. 3 is a schematic structural diagram of a cylindrical mirror according to the present invention;

fig. 4 is a schematic structural view of an excitation light filter wheel or an emission light filter wheel according to the present invention;

fig. 5 is a linear light spot incident to the TDI line camera according to the present invention;

fig. 6 is an illuminance distribution curve of a light spot according to embodiment 1 of the present invention.

Detailed Description

The present invention is described in detail below, so that the advantages and features of the present invention can be more easily understood by those skilled in the art, thereby making more clear and definite definitions of the protection scope of the present invention.

The following presents a simplified summary of one or more aspects in order to provide a basic understanding of such aspects. This summary is not an extensive overview of all contemplated aspects, and is intended to neither identify key or critical elements of all aspects nor delineate the scope of any or all aspects. Its sole purpose is to present some concepts of one or more aspects in a simplified form as a prelude to the more detailed description that is presented later.

In the description of the present invention, it is to be understood that the terms "upper", "lower", "front", "rear", "left", "right", "top", "bottom", "inner", "outer", and the like indicate orientations or positional relationships based on the orientations or positional relationships shown in the drawings, and are only for convenience of description and simplicity of description, and do not indicate or imply that the device or element being referred to must have a particular orientation, be constructed and operated in a particular orientation, and therefore, should not be construed as limiting the present invention.

As shown in fig. 1 to 6, a line scanning fluorescence microscope using a multi-wavelength laser light source includes a beam combiner, a powell prism 10, a cylindrical mirror 11, an excitation light filter wheel 12, a beam splitter 13, a microscope objective 14, an emission light filter wheel 16, a tube lens 17, and a TDI line camera 18, the laser group, the collimating lens group 5, and the dichroic mirror group constitute the beam combiner, the laser group, the collimating lens group, the dichroic lens group, the powell prism 10, the cylindrical mirror 11, the excitation light filter wheel 12, and the beam splitter 13 are sequentially arranged along a light incidence direction, the microscope objective 14 and a sample 15 to be measured are disposed on the same side of the beam splitter 13, the tube lens 17 and the TDI line camera 18 are disposed on the other side of the beam splitter 13, a light beam emitted from the excitation light filter wheel 12 can be incident on the microscope objective 14, the light beam is irradiated on the sample 15 to be measured through the microscope objective 14 to excite fluorescence, the fluorescence is reflected with a portion of the excitation light and then is incident on the beam splitter 13 through the tube lens 14, the emission light emitted from the beam splitter 13 is filtered, only the excitation light is allowed to be transmitted through the tube filter wheel 16, and then incident on the tube lens 17 of the fluorescence tube lens 17.

The laser group comprises a plurality of lasers with different wavelengths, the wavelength range is 330 nm-550 nm, the collimating lens group comprises a plurality of collimating lenses, the collimating lenses can adopt spherical lenses, aspheric lenses, cylindrical lenses or spherical lenses, an antireflection film is plated on the collimating lenses to reduce the loss of optical power, the dichroic mirror group comprises a plurality of dichroic mirrors, the number of the lasers, the collimating lenses and the dichroic mirrors is the same, each laser corresponds to one collimating lens and one dichroic mirror, and the laser emitted by each laser is emitted as collimated light after passing through the corresponding collimating lens, is emitted through the corresponding dichroic mirror and then enters the Baville prism 10 after being emitted and combined.

The light spots emitted by the Powell prism 10 are linear light spots, strict requirements are imposed on the diameter of the light spots incident on the prism, and if the diameter of the incident light spots is not matched with the required diameter, the illumination intensity of the emitted light spots is not uniform.

The excitation light filter wheel 12 and the emission light filter wheel 16 have the same structure and are disposed at different angles. Taking the excitation light filter wheel 12 as an example to illustrate the structure, the excitation light filter wheel 12 includes a panel, a rotating motor M and a filter wheel, four filters with the same diameter are uniformly distributed on the panel, the filter wheel is controlled by the rotating motor M, and when the excitation light (fluorescence) with different wavelengths is used, the rotating motor M drives the filter wheel to rotate to the corresponding filter.

The utility model discloses a theory of operation does:

laser emitted by a laser group is converted into collimated light through a collimating lens group and emitted, the collimated light is combined into a beam of light through a dichroic lens group, the diameter of the combined light spot can be 0.8-3 mm, the combined light is incident on a Powell prism 10, the diameter of the light spot incident on the Powell prism 10 is consistent with the diameter of the incident light spot required by the prism, the diameter of the light spot incident on the Powell prism is usually required to be 0.8-3 mm, an antireflection film is plated on the surface of the Powell prism 10, the fan angle of the Powell prism 10 can be 10-60 degrees, the light spot emitted by the Powell prism 10 is a linear light spot with uniform illumination, the beam emitted by the Powell prism 10 is incident on a cylindrical mirror 11, the surface of the cylindrical mirror 11 is plated with an antireflection film, the cylindrical mirror 11 plays roles in focusing and changing the divergence angle of the beam, the beam emitted by the cylindrical mirror 11 passes through an excitation light filter wheel 12 to filter other light with specific wavelength, and the beam emitted by the excitation light filter wheel 12 is divided into two beams through a beam splitter 13, one light beam is incident into the microscope objective 14, the divergence angle of the light beam is matched with the view field of the microscope objective 14, the line view field of the microscope objective 14 is 10-30 mm, the numerical aperture NA is 0.1-1.25, the magnification is 4X-100X, the light beam irradiates on the sample 15 to be measured through the microscope objective 14 to excite fluorescence, the fluorescence and part of excitation light are reflected, so that the light reversely passes through the microscope objective lens 14 and then enters the beam splitter 13, the light emitted from the beam splitter 13 passes through the emission light filter wheel 16 to filter the excitation light, only the fluorescence is allowed to transmit, the fluorescence enters the tube lens 17 again, the tube lens 17 should be used in cooperation with the microscope objective 14, and usually the focal length of the tube lens 17 is 180 mm-200 mm, the linear field of view is 10-30 mm, and the fluorescence passing through the tube lens 17 is finally incident into the TDI line camera 18.

Example 1

As shown in fig. 1 to 6, a line scanning fluorescence microscope using a multi-wavelength laser light source includes a laser group, a collimating lens group 5, a dichroic lens group, a powell prism 10, a cylindrical lens 11, an excitation light filter wheel 12, a beam splitter 13, a microscope objective 14, an emission light filter wheel 16, a tube lens 17, and a TDI line camera 18, the laser group, the collimating lens group, the dichroic lens group, the powell prism 10, the cylindrical lens 11, the excitation light filter wheel 12, and the beam splitter 13 are sequentially arranged along a light incidence direction, the microscope objective 14 and a sample 15 to be measured are arranged on the same side of the beam splitter 13, the other side of the beam splitter 13 is provided with the tube lens 17 and the TDI line camera 18, a light beam emitted through the excitation light filter wheel 12 can be incident on the microscope objective 14 through the beam splitter 13, the light beam is irradiated onto the sample 15 to be measured through the microscope objective 14 to excite fluorescence, the fluorescence is reflected with a part of the excitation light and then enters the microscope objective 13 through the microscope objective 14 in a reverse direction, the emission light emitted from the beam splitter 13, and then is filtered, only the excitation light is allowed to be transmitted through the tube lens 17, and finally enters the TDI line camera 18.

The laser group includes four lasers with different wavelengths, and the four lasers are sequentially recorded as follows from left to right: the device comprises a laser I1, a laser II 2, a laser III 3 and a laser IV 4; the collimating lens group 5 comprises four collimating lenses which are sequentially arranged from left to right; dichroic mirror group includes four dichroic mirrors, marks in proper order from left to right: a dichroic mirror I6, a dichroic mirror II 7, a dichroic mirror III 8 and a dichroic mirror IV 9; each laser corresponds to one collimating lens and one dichroic mirror, that is: the laser I1, the leftmost collimating lens and the dichroic mirror I6 are in one-to-one correspondence and adaptation in position, the laser I1 is collimated light and emitted after passing through the leftmost collimating lens, the collimated light and the emitted light pass through the dichroic mirror I6, the laser IV 4, the rightmost collimating lens and the dichroic mirror IV 9 are in one-to-one correspondence and adaptation in position, the laser IV 4 is collimated light and emitted after passing through the rightmost collimating lens, the collimated light and the emitted light pass through the dichroic mirror IV 9 and are emitted, the laser II 2, the middle left collimating lens and the dichroic mirror II 7 are in one-to-one correspondence and adaptation in position, the laser II 2 is collimated light and emitted after passing through the middle left collimating lens, the laser II is emitted through the dichroic mirror II 7 and is emitted, the laser III 3, the middle right collimating lens and the dichroic mirror III 8 are in one-to-one correspondence and adaptation in position, the laser III 3 passes through the middle right collimating lens and is emitted as collimated light and emitted through the dichroic mirror III 8, the laser is emitted as a spot with a strict exit requirement that the exit diameter of the exit spot on the Babye-Boerhan exit prism, and the exit diameter of the incident light spot on the Babye-Boerhan incident prism is uneven.

The excitation light filter wheel 12 and the emission light filter wheel 16 have the same structure and are disposed at different angles. Taking the excitation light filter wheel 12 as an example to illustrate the structure, the excitation light filter wheel 12 includes a panel, a rotating motor M and a filter wheel, four filters with the same diameter are uniformly distributed on the panel, the filter wheel is controlled by the rotating motor M, and when excitation light (fluorescence) with different wavelengths is used, the rotating motor M drives the filter wheel to rotate to the corresponding filter.

The utility model discloses a theory of operation does:

laser emitted by a laser group is converted into collimated light through a collimating lens group and emitted, the collimated light is combined into a beam of light through a dichroic lens group, the diameter of a combined light spot is 0.8mm, the combined light is emitted onto a Powell prism 10, the diameter of the light spot emitted onto the Powell prism 10 is consistent with that of an incident light spot required by the Powell prism, the diameter of the light spot emitted onto the Powell prism is generally required to be 0.8mm, an antireflection film is coated on the surface of the Powell prism 10, the fan angle of the Powell prism 10 is 60 degrees, the light spot emitted out through the Powell prism 10 is a linear light spot with uniform illumination, the light beam emitted out through the Powell prism 10 is emitted onto a cylindrical lens 11, the surface of the cylindrical lens 11 is coated with an antireflection film, the cylindrical lens 11 plays a role in focusing and changing the divergence angle of the light beam, the light beam emitted through the cylindrical lens 11 is filtered by an excitation light filter wheel 12 to filter other light with a specific wavelength, the light beam emitted through the excitation light filter wheel filter 12 is divided into two beams, one beam 14, wherein the one beam 14 is emitted into a microscope objective lens 14, the microscope objective 14, the emission light is emitted from a fluorescence microscope objective lens barrel 17, and is emitted into a fluorescence microscope objective lens 14, and then emitted into a fluorescence microscope objective lens 17, and emitted light to be reflected by a fluorescence microscope objective lens 17 to be emitted.

As can be seen from fig. 6, the illuminance of the light spot obtained by the line scanning fluorescence microscope using the multi-wavelength laser light source of the present embodiment is relatively uniform, and the requirement of fluorescence illumination can be met.

The utility model discloses the part or the structure that do not specifically describe adopt prior art or current product can, do not do here and describe repeatedly.

The above only is the embodiment of the present invention, not limiting the patent scope of the present invention, all utilize the equivalent structure or equivalent flow transformation that the content of the specification does, or directly or indirectly use in other related technical fields, all including in the same way the patent protection scope of the present invention.

Claims (10)

1. A line scanning type fluorescence microscope using a multi-wavelength laser light source is characterized by comprising a beam combination optical group, a Powell prism, a cylindrical mirror, an exciting light filter wheel, a beam splitter, a microscope objective, an emitting light filter wheel, a tube lens and a TDI linear array camera, wherein the beam combination optical group, the Powell prism, the cylindrical mirror, the exciting light filter wheel and the beam splitter are sequentially arranged along a light incidence direction, the microscope objective is arranged on one side of the beam splitter, and the tube lens and the TDI linear array camera are arranged on the other side of the beam splitter.

2. The line-scanning fluorescence microscope using a multi-wavelength laser light source according to claim 1, wherein the combined beam light group includes a laser group, a collimating lens group and a dichroic lens group, which are sequentially arranged along a light incidence direction, the laser group includes a plurality of lasers having different wavelengths, the collimating lens group includes a plurality of collimating lenses, the dichroic lens group includes a plurality of dichroic mirrors, the number of lasers, collimating lenses and dichroic mirrors is the same, and each laser corresponds to one collimating lens and one dichroic mirror.

3. The line scanning fluorescence microscope using multi-wavelength laser light source as claimed in claim 2, wherein the laser set comprises four lasers with different wavelengths, which are sequentially recorded as follows from left to right: the device comprises a laser I, a laser II, a laser III and a laser IV; the collimating lens group comprises four collimating lenses which are sequentially arranged from left to right; dichroic mirror group includes four dichroic mirrors, records as in proper order from left to right: a dichroic mirror I, a dichroic mirror II, a dichroic mirror III and a dichroic mirror IV; the laser device I, the leftmost collimating lens and the dichroic mirror I are in one-to-one correspondence and adaptation in position, the laser device II, the middle left collimating lens and the middle right collimating lens are in one-to-one correspondence and adaptation in position, the laser device III, the middle right collimating lens and the dichroic mirror III are in one-to-one correspondence and adaptation in position, and the laser device IV, the rightmost collimating lens and the dichroic mirror IV are in one-to-one correspondence and adaptation in position.

4. The line scanning fluorescence microscope using the multi-wavelength laser light source as claimed in claim 2, wherein the laser has a wavelength ranging from 330nm to 550nm.

5. The line scanning fluorescence microscope of claim 2, wherein the collimating lens is a spherical lens, an aspherical lens, a cylindrical lens or a spherical lens, and the collimating lens is coated with an antireflection film.

6. The fluorescence microscope of claim 2, wherein the dichroic mirror group has an exit spot diameter of 0.8-3 mm, and the incident spot diameter on the Powell prism is 0.8-3 mm.

7. The line scanning fluorescence microscope using the multi-wavelength laser light source as claimed in claim 1, wherein the surface of the Powell prism is coated with an antireflection film, the sector angle of the Powell prism is 10-60 °, and the light spot emitted from the Powell prism is a linear light spot.

8. The line scanning fluorescence microscope using the multi-wavelength laser light source as claimed in claim 1, wherein the microscope objective has a line field of view of 10 to 30mm, a numerical aperture NA of 0.1 to 1.25, and a magnification of 4X to 100X.

9. The line scanning fluorescence microscope using a multi-wavelength laser light source according to claim 1, wherein the tube lens has a focal length of 180mm to 200mm and a line field of view of 10 mm to 30mm.

10. The line scanning fluorescence microscope of claim 1, wherein the excitation light filter wheel and the emission light filter wheel are identical in structure and each comprise a panel, a rotating motor M and a filter wheel, four filters with the same diameter are uniformly distributed on the panel, and the filter wheel is connected with and controlled by the rotating motor M.

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