An oblique phase contrast imaging device for an inverted biological microscope

Abstract

The present invention discloses an oblique phase contrast imaging device for an inverted biological microscope, belonging to the field of optical microscopic imaging technology. The device comprises an oblique phase contrast imaging plate disposed below the condenser of the inverted biological microscope, and an oblique phase contrast light source disposed on a frame support surface below the stage of the inverted biological microscope. The light beam emitted by the oblique phase contrast light source is emitted onto the oblique phase contrast imaging plate via a wave plate on the stage of the inverted biological microscope. The present invention solves the problem of poor performance and complex operation when observing brightfield fluorescence using an inverted biological microscope.

Description

Technical Field

[0001] The invention belongs to the technical field of optical microscopic imaging, and in particular relates to an oblique phase contrast imaging device for an inverted biological microscope. Background Art

[0002] Inverted biological microscopes are often used for cell observation. Because cells are thin and transparent, conventional brightfield observations struggle to reveal internal details. Traditional inverted biological microscopes employ imaging techniques such as phase contrast, differential interference contrast, and Hoffman imaging to render cells more three-dimensional while enhancing cell contrast and detail. Phase contrast is the most widely used and cost-effective method.

[0003] Phase contrast imaging requires the use of a phase contrast objective lens and its accompanying phase ring. Adjusting the ring's center position during use is inconvenient. Furthermore, when observing brightfield fluorescence using a phase contrast objective lens, the effect is affected by the phase ring within the objective lens, resulting in reduced clarity and brightness. Summary of the Invention

[0004] In response to the above-mentioned deficiencies in the prior art, the present invention provides an oblique phase contrast imaging device for an inverted biological microscope. By arranging an oblique phase contrast imaging plate and an oblique phase contrast light source on an inverted biological microscope, the problem of poor effect and complicated operation when using an inverted biological microscope to observe fluorescence in bright field is solved.

[0005] In order to achieve the above-mentioned object of the invention, the technical solution adopted by the present invention is:

[0006] The present invention provides an oblique phase contrast imaging device for an inverted biological microscope, comprising an oblique phase contrast imaging plate disposed below a condenser of the inverted biological microscope, and an oblique phase contrast light source disposed on a frame bearing surface below the stage of the inverted biological microscope;

[0007] The light beam emitted by the oblique phase contrast light source is emitted onto the oblique phase contrast imaging plate through the wave plate on the stage of the inverted biological microscope.

[0008] Furthermore, the angle between the oblique phase-contrast light source and the rack bearing surface below the stage of the inverted biological microscope is 50-60 degrees, and the projection distance between the light beam emission center of the oblique phase-contrast light source and the center of the objective lens turret is 80-90 mm.

[0009] Furthermore, the oblique phase difference light source includes lamp beads, plano-convex lenses and biconvex lenses whose centers are on a straight line;

[0010] The light beam emitted by the lamp bead is incident on the plane of the plano-convex lens, passes through the internal medium of the plano-convex lens, and is refracted from the convex surface of the plano-convex lens; the light beam refracted from the convex surface of the plano-convex lens is refracted and incident on the first convex surface of the biconvex lens, passes through the internal medium of the biconvex lens, and is refracted from the second convex surface of the biconvex lens; the light beam refracted from the second convex surface of the biconvex lens is emitted onto the oblique phase difference imaging plate through the wave plate on the stage of the inverted biological microscope.

[0011] Furthermore, the oblique phase difference light source further includes an assembly shell;

[0012] A pressure ring is provided in the assembly shell to lock the center positions of the lamp bead, the plano-convex lens and the biconvex lens in a straight line, wherein the shortest distance between the outer side of the lamp bead and the first convex surface of the biconvex lens is 30 mm.

[0013] Furthermore, a guide groove is provided on the assembly shell, in which a drag rod fixedly connected to the plano-convex lens is provided; by dragging the drag rod in the guide groove, the relative position of the plano-convex lens between the lamp bead and the double convex lens can be adjusted as the drag rod moves.

[0014] Furthermore, the oblique phase difference imaging plate adopts a light homogenizing plate or a black board.

[0015] The beneficial effects of the present invention are as follows: the present invention provides an oblique phase contrast imaging device for an inverted biological microscope, which realizes the adjustment of the position and angle of the illumination light source by adding an oblique phase contrast imaging plate and an oblique phase contrast light source on the basis of an ordinary inverted biological microscope, so that transparent or fluorescent observation objects such as cells can present a three-dimensional effect with similar phase contrast; the oblique phase contrast imaging device provided by the present invention can observe the three-dimensional effect of cells only by using a conventional bright field objective lens, without affecting the effects under bright field and fluorescence, and does not require adjusting the phase contrast ring. Compared with the cumbersome operation of replacing and adjusting the phase contrast ring of the traditional phase contrast objective lens, it is more convenient to use; the oblique phase contrast light source provided by the present invention can adjust the output light beam by dragging the drag rod to achieve the oblique phase contrast effect of objective lenses of different magnifications. After being matched with the oblique phase contrast imaging plate, it can not only be used to observe culture dish cells, but also has the same three-dimensional effect on cell slides.

[0016] Other advantages of the present invention will be analyzed in more detail in subsequent embodiments. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following is a brief introduction to the drawings required for use in the embodiments. It should be understood that the following drawings only illustrate certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other relevant drawings can be obtained based on these drawings without paying any creative work.

[0018] Figure 1 Schematic diagram of the structure of an oblique phase contrast imaging device for an inverted biological microscope according to an embodiment of the present invention.

[0019] Figure 2 Schematic diagram of the relative positions of the oblique phase difference light source and the objective lens turret in an embodiment of the present invention.

[0020] Figure 3 2 is a light path diagram of an oblique phase difference light source in an embodiment of the present invention.

[0021] Figure 4 Schematic diagram of the assembly housing, guide groove and drag rod of the oblique phase difference light source in an embodiment of the present invention.

[0022] FIG5( a ) is an image of a cell slide observed with an inverted biological microscope at 10x magnification in bright field according to an embodiment of the present invention.

[0023] FIG5( b ) is an image of a cell slide observed with an inverted biological microscope at 20 times magnification in bright field according to an embodiment of the present invention.

[0024] FIG5( c ) is an image of a cell slide observed at 10 times magnification using an oblique phase contrast imaging device combined with an inverted biological microscope in an embodiment of the present invention.

[0025] FIG5( d ) is an image of a cell slide observed at 20 times magnification using an oblique phase contrast imaging device combined with an inverted biological microscope in an embodiment of the present invention.

[0026] FIG6( a ) is an image of cells in a culture dish observed using an inverted biological microscope at 10x magnification in bright field according to an embodiment of the present invention.

[0027] FIG6( b ) is an image of cells in a culture dish observed using an inverted biological microscope at 20 times magnification in bright field according to an embodiment of the present invention.

[0028] FIG6( c ) is an image of cells in a culture dish observed using an oblique phase contrast imaging device combined with an inverted biological microscope at 10 times magnification in an embodiment of the present invention.

[0029] FIG6( d ) is an image of cells in a culture dish observed at 20 times magnification using an oblique phase contrast imaging device combined with an inverted biological microscope in an embodiment of the present invention.

[0030] Among them: 1. Condenser; 2. Oblique phase contrast imaging plate; 3. Stage; 4. Rack bearing surface; 5. Oblique phase contrast light source; 501. Lamp beads; 502. Plano-convex lens; 503. Biconvex lens; 504. Assembly shell; 505. Guide groove; 506. Drag rod; 6. Objective turret; 7. Wave plate. DETAILED DESCRIPTION

[0031] The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, not all of the embodiments. The components of the embodiments of the present invention generally described and shown in the drawings herein can be arranged and designed in various different configurations. Therefore, the following detailed description of the embodiments of the present invention provided in the drawings is not intended to limit the scope of the claimed invention, but merely represents selected embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without making creative work are within the scope of protection of the present invention.

[0032] like Figure 1 As shown, in one embodiment of the present invention, an oblique phase contrast imaging device for an inverted biological microscope is provided, comprising an oblique phase contrast imaging plate 2 disposed below a condenser lens 1 of the inverted biological microscope, and an oblique phase contrast light source 5 disposed on a frame bearing surface 4 below a stage 3 of the inverted biological microscope;

[0033] The light beam emitted by the oblique phase contrast light source 5 is emitted onto the oblique phase contrast imaging plate 2 through the wave plate 7 on the stage 3 of the inverted biological microscope.

[0034] In this embodiment, the distance from the center of the condenser 1 of the inverted biological microscope to the lower edge of the stage 3 is 55 mm.

[0035] like Figure 2 As shown, the angle between the oblique phase difference light source 5 and the frame bearing surface 4 below the stage 3 of the inverted biological microscope is 50-60 degrees, and the projection distance between the light beam emission center of the oblique phase difference light source 5 and the center of the objective lens turret 6 is 80-90 mm.

[0036] like Figure 3 As shown, the oblique phase difference light source 5 includes a lamp bead 501, a plano-convex lens 502 and a biconvex lens 503 whose centers are on a straight line;

[0037] The light beam emitted by the lamp bead 501 is incident on the plane of the plano-convex lens 502, and after passing through the internal medium of the plano-convex lens 502, it is refracted and emitted from the convex surface of the plano-convex lens 502; the light beam refracted and emitted from the convex surface of the plano-convex lens 502 is refracted and incident on the first convex surface of the biconvex lens 503, and after passing through the internal medium of the biconvex lens 503, it is refracted and emitted from the second convex surface of the biconvex lens 503; the light beam refracted and emitted from the second convex surface of the biconvex lens 503 is emitted onto the oblique phase difference imaging plate 2 through the wave plate 7 on the stage 3 of the inverted biological microscope.

[0038] like Figure 4 As shown, the oblique phase difference light source 5 further includes an assembly shell 504;

[0039] A pressure ring is provided in the assembly shell 504 to lock the center positions of the lamp bead 501, the plano-convex lens 502 and the biconvex lens 503 in a straight line, wherein the shortest distance between the outer side of the lamp bead 501 and the first convex surface of the biconvex lens 503 is 30 mm.

[0040] In this solution, the assembly housing 504 aligns the centers of the lamp 501, plano-convex lens 502, and biconvex lens 503, ensuring stable operation of the oblique phase-contrast light source 5. Furthermore, the light beam generated by the lamp 501, after being regulated by the plano-convex lens 502 and biconvex lens 503, facilitates observation of cells and other structures using an inverted biological microscope. The assembly housing 504 also effectively protects the optical path components of the oblique phase-contrast light source 5.

[0041] The assembly shell 504 is also provided with a guide groove 505, in which a drag rod 506 fixedly connected to the plano-convex lens 502 is provided; by dragging the drag rod 506 to move in the guide groove 505, the relative position of the plano-convex lens 502 between the lamp bead 501 and the biconvex lens 503 can be adjusted as the drag rod 506 moves.

[0042] In this solution, the dragging of the drag rod 506 is mainly determined by the height of the stage 3 and the imaging effect. By adjusting the relative position between the plano-convex lens 502, the lamp bead 501 and the biconvex lens 503, the angle of the light beam emitted from the oblique phase difference light source 5 can be adjusted, so that the emitted light beam can achieve the oblique phase difference effect of objective lenses of different magnifications, so as to adapt to more different observation samples.

[0043] In this embodiment, when the drag rod 506 is dragged in the guide groove 505 to move toward the double convex lens 503, the plano-convex lens 502 moves in the assembly shell 504 along with the drag rod 506 toward the double convex lens 503, that is, the relative position between the plano-convex lens 502 and the double convex lens 503 becomes closer, and the relative position between the plano-convex lens 502 and the lamp bead 501 becomes farther, so that the light emitted by the lamp bead 501 is more divergent; when the drag rod 506 is dragged in the guide groove 505 to move toward the lamp bead 501, the plano-convex lens 502 moves in the assembly shell 504 along with the drag rod 506 toward the lamp bead 501, that is, the relative position between the plano-convex lens 502 and the lamp bead 501 becomes closer, and the relative position between the plano-convex lens 502 and the double convex lens 503 becomes farther, so that the light emitted by the lamp bead 501 is more convergent.

[0044] The oblique phase difference imaging plate 2 is a light homogenizing plate or a blackboard.

[0045] In this solution, different materials of oblique phase contrast imaging plates 2 are selected according to the different objects of observation, which can achieve better coordinated observation effects. In this embodiment, a light-diffusing plate is used for cell slide observation, and a blackboard is used for culture dish cell observation. The light-diffusing plate usually only plays the role of light uniformity, while in this solution, the light-diffusing plate scatters or reflects light to better achieve oblique phase contrast imaging for cell slide observation. Similarly, the blackboard usually only plays the role of absorbing and blocking light, while in this solution, the light-diffusing plate scatters or reflects light to better achieve oblique phase contrast imaging for culture dish cell observation. When observing cell slides and culture dish cells, samples are placed on the wave plate 7 and observed through an inverted microscope.

[0046] The oblique phase contrast imaging device for an inverted biological microscope provided by the present invention can enable cells that are originally opaque under bright field conditions to present a stereoscopic imaging effect similar to differential interference observation under the assistance of an oblique phase contrast light source 5 in combination with an oblique phase contrast baffle, and does not require the corresponding phase contrast objective lens and corresponding phase contrast ring required by traditional phase contrast.

[0047] The working principle of the present invention is as follows: the lamp bead 501 of the oblique phase difference light source 5 is energized, so that the oblique phase difference light source 5 emits a light beam, the light beam emitted by the lamp bead 501 is incident on the plane of the plano-convex lens 502, passes through the internal medium of the plano-convex lens 502, and is refracted from the convex surface of the plano-convex lens 502; the light beam refracted from the convex surface of the plano-convex lens 502 is refracted and incident on the first convex surface of the biconvex lens 503, passes through the internal medium of the biconvex lens 503, and is refracted from the second convex surface of the biconvex lens 503; the light beam refracted from the second convex surface of the biconvex lens 503 is emitted onto the oblique phase difference imaging plate 2 through the wave plate 7 on the stage 3 of the inverted biological microscope. When the oblique phase contrast imaging device for an inverted biological microscope provided by the present invention is working, a cell slide or culture dish cells are provided on the wave plate 7 of the stage 3. If the object of observation is a cell slide, a homogenizing plate is used as the oblique phase contrast imaging plate 2. If the object of observation is a culture dish cells, a blackboard is used as the oblique phase contrast imaging plate 2. When observing the cell slide or culture dish cells, the light beam emitted from the oblique phase contrast light source 5 can be adjusted by simply dragging the drag rod 506, so that when observing transparent or fluorescent objects, such as cells, through an inverted biological microscope based on the emitted light beam, the oblique phase contrast effect of objective lenses of different magnifications can be achieved.

[0048] As shown in FIG5(a) and FIG5(b), in a practical example of the present invention, a cell slide is directly observed using an inverted biological microscope under 10x bright field and 20x bright field conditions, and no obvious three-dimensional features are observed in the cell slide. However, after using the oblique phase contrast device proposed in the present invention in combination with an inverted biological microscope, as shown in FIG5(c) and FIG5(d), a light homogenizing plate is used under 10x magnification and 20x magnification conditions, and the light beam emitted from the oblique phase contrast light source is adjusted by dragging the drag rod 506, so that the three-dimensional features of the cell slide can be effectively observed, and the imaging effect is similar to that of differential interference contrast imaging.

[0049] As shown in FIG6 (a) and FIG6 (b), in a practical example of the present invention, the culture dish cells are directly observed using an inverted biological microscope under 10x bright field and 20x bright field conditions, and it is observed that the culture dish cells have no obvious three-dimensional features. However, after using the oblique phase contrast device proposed by the present invention in combination with an inverted biological microscope, as shown in FIG6 (c) and FIG6 (d), a blackboard is used under 10x magnification and 20x magnification conditions, and the three-dimensional features of the culture dish cells can be effectively observed by dragging the drag rod 506 to adjust the light beam emitted from the oblique phase contrast light source. The imaging effect is similar to that of differential interference contrast imaging.

[0050] The above description is only a specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any technician familiar with this technical field can easily think of changes or replacements within the technical scope disclosed by the present invention, which should be covered by the scope of protection of the present invention.

Claims

1. An oblique phase contrast imaging device for an inverted biological microscope, characterized in that: It comprises an oblique phase contrast imaging plate (2) arranged below the condenser (1) of the inverted biological microscope, and an oblique phase contrast light source (5) arranged on a frame bearing surface (4) below the stage (3) of the inverted biological microscope; The light beam emitted by the oblique phase-contrast light source (5) is emitted onto the oblique phase-contrast imaging plate (2) via the wave plate (7) on the stage (3) of the inverted biological microscope; The angle between the oblique phase-contrast light source (5) and the frame bearing surface (4) below the stage (3) of the inverted biological microscope is 50 to 60 degrees, and the projection distance between the light beam emission center of the oblique phase-contrast light source (5) and the center of the objective lens turntable (6) is 80 to 90 mm; The oblique phase difference light source (5) comprises a lamp bead (501) whose center positions are on a straight line, a plano-convex lens (502) and a biconvex lens (503); The light beam emitted by the lamp bead (501) is incident on the plane of the plano-convex lens (502), passes through the internal medium of the plano-convex lens (502), and is refracted from the convex surface of the plano-convex lens (502); the light beam refracted from the convex surface of the plano-convex lens (502) is refracted and incident on the first convex surface of the biconvex lens (503), passes through the internal medium of the biconvex lens (503), and is refracted from the second convex surface of the biconvex lens (503); the light beam refracted from the second convex surface of the biconvex lens (503) is emitted onto the oblique phase difference imaging plate (2) via the wave plate (7) on the stage (3) of the inverted biological microscope; The oblique phase difference light source (5) further includes an assembly shell (504); A pressure ring is provided in the assembly shell (504), so that the center positions of the lamp bead (501), the plano-convex lens (502), and the biconvex lens (503) are locked in a straight line, wherein the shortest distance between the outer side of the lamp bead (501) and the first convex surface of the biconvex lens (503) is 30 mm; The assembly shell (504) is further provided with a guide groove (505), in which a drag rod (506) fixedly connected to the plano-convex lens (502) is provided; when the drag rod (506) is dragged to move in the guide groove (505), the relative position of the plano-convex lens (502) between the lamp bead (501) and the biconvex lens (503) can be adjusted along with the movement of the drag rod (506).

2. The oblique phase contrast imaging device for an inverted biological microscope according to claim 1, characterized in that: The oblique phase difference imaging plate (2) adopts a light homogenizing plate or a blackboard.

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