CN109633883B - Large-view-field high-resolution fluorescence microscope objective lens - Google Patents
Large-view-field high-resolution fluorescence microscope objective lens Download PDFInfo
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- CN109633883B CN109633883B CN201910063663.0A CN201910063663A CN109633883B CN 109633883 B CN109633883 B CN 109633883B CN 201910063663 A CN201910063663 A CN 201910063663A CN 109633883 B CN109633883 B CN 109633883B
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Abstract
The invention discloses a large-field-of-view high-resolution fluorescence microscope objective lens, which comprises a glass slide S, a dichroic mirror D and an emission optical filter E, wherein the glass slide S, the dichroic mirror D and the emission optical filter E are sequentially arranged from an object plane to an image plane; a lens or a lens group is placed on at least one of the position between the glass slide S and the dichroic mirror D, the position between the dichroic mirror D and the emission filter E and the position behind the emission filter E; the obtained fluorescence microscope objective has the magnification of 10-35 times, the numerical aperture of 0.3-0.5, the wavelength visible light range of 480nm-680nm, the object space view field diameter of 4mm-10mm and the object space resolution of 0.4-2 μm. The MTF value of the fluorescence microscope objective lens is close to the diffraction limit in the working waveband full-field range, the relative distortion is less than 0.3% in the working waveband full-field range, and compared with the traditional objective lens with the same field, the fluorescence microscope objective lens has higher imaging resolution and smaller relative distortion.
Description
Technical Field
The invention belongs to the field of optics, and particularly relates to a large-field-of-view high-resolution fluorescence microscope objective.
Background
Microscope objectives with large numerical apertures are used for examining biological objects and tissue structures in order to resolve fine structures. However, it is difficult for existing microscopes (conventional optical microscopes, electron microscopes, confocal microscopes, and even the most advanced two-photon microscopes in recent years), cameras and cameras, and living body imaging apparatuses to realize wide-field imaging at high resolution. To obtain a wide field of view image at high resolution, the aperture of the imaging optical system must necessarily be increased. However, due to the influence of the off-axis aberration of the lens, the edge and the center magnification ratio are not matched in the multi-lens imaging process, so that the distortion of the edge of the wide field of view is large. At present, the objective lens has inherent restriction on large visual field and high resolution index, cannot be optimized simultaneously, cannot meet the research requirement of life science, and needs to be improved and broken through urgently.
Disclosure of Invention
The invention aims to overcome the defects in the prior art and provides a fluorescence microscope objective with large field of view and high resolution.
In order to achieve the purpose, the invention adopts the following technical scheme: a large-field-of-view high-resolution fluorescence microscope objective comprises a glass slide S, a dichroic mirror D and an emission filter E which are sequentially arranged from an object plane to an image plane; a lens or a lens group is placed on at least one of the position between the glass slide S and the dichroic mirror D, the position between the dichroic mirror D and the emission filter E and the position behind the emission filter E; the obtained fluorescence microscope objective has the magnification of 10-35 times, the numerical aperture of 0.3-0.5, the wavelength visible light range of 480nm-680nm, the object space view field diameter of 4mm-10mm and the object space resolution of 0.4-2 μm.
Further, the fluorescence microscope objective lens comprises a glass slide S, a medium W, a collimating lens group G1, a reverse telelens group G2, a dichroic mirror D, an emission filter E, an achromatic lens group G3 and a Gaussian-like lens group G4 which are sequentially arranged from the object plane to the image plane.
Further, the collimating lens group G1 includes a first plano-convex lens L1, a first convex-concave lens L2, a first concave-convex lens L3, and a second plano-convex lens L4, which are sequentially arranged along the optical axis direction.
Further, the anti-telephoto group G2 includes a first biconcave lens L5, a first biconvex lens L6, a second biconvex lens L7, and a second convex-concave lens L8, which are sequentially arranged in the optical axis direction.
Further, the achromatic lens group G3 includes a third biconvex lens L9 and a third convex-concave lens L10 arranged in this order in the optical axis direction.
Further, the gauss-like lens group comprises a fourth convex-concave lens L11, a second double-concave lens L12, a fifth convex-concave lens L13, a second convex-concave lens L14 and a third convex-concave lens L15 which are sequentially arranged along the optical axis direction, and the fluorescence microscope objective lens adopts a gauss-like lens group G4 deformed by the gauss lens group to shorten the object image conjugate distance.
Furthermore, the principal rays of the incident end of the dichroic mirror and the emergent end of the emission filter are strictly parallel, and the lens group of the incident end of the dichroic mirror and the emergent end of the emission filter retains aberration to compensate the aberration generated by the two flat plates.
Furthermore, the image surface of the fluorescence microscope objective is a curved surface.
Further, the fluorescence microscope objective lens gives consideration to both object-side telecentricity and image-side telecentricity.
Furthermore, the initial structure of the fluorescence microscope objective adopts a form which mainly takes the structure of the microscope objective into consideration the structural characteristics of the photoetching objective.
The invention has the following beneficial effects: through computer-aided optical design and optimization, the number and the structure of the lens groups are reasonably selected, each lens is of a spherical surface type and is coaxially arranged, the processing and the assembly and the adjustment are convenient, the lens materials are common commercial glass, and the purchasing difficulty and the manufacturing cost of the optical system materials are reduced. The MTF value of the fluorescence microscope objective lens is close to the diffraction limit in the working waveband full-field range, the relative distortion is less than 0.3% in the working waveband full-field range, and compared with the traditional objective lens with the same field, the fluorescence microscope objective lens has higher imaging resolution and smaller relative distortion.
Drawings
FIG. 1 is a diagram of the optical system of a fluorescence microscope objective according to a preferred embodiment of the present invention;
FIG. 2 is an enlarged view of a portion of FIG. 1;
FIG. 3 is a diagram of the modulation transfer function of the objective lens of a fluorescence microscope according to a preferred embodiment of the present invention;
FIG. 4 is a schematic diagram of a fluorescence microscope objective according to a preferred embodiment of the present invention;
FIG. 5 is a graph of field curvature and distortion of a fluorescence microscope objective lens according to a preferred embodiment of the present invention;
FIG. 6 is a diagram of the optical system of the objective lens of a fluorescence microscope according to another preferred embodiment of the present invention;
FIG. 7 is an enlarged view of a portion of FIG. 6;
FIG. 8 is a diagram of the modulation transfer function of a fluorescence microscope objective lens according to another preferred embodiment of the present invention;
FIG. 9 is a dot-column diagram of a fluorescence microscope objective according to another preferred embodiment of the present invention;
FIG. 10 is a graph of field curvature and distortion of a fluorescence microscope objective lens according to another preferred embodiment of the present invention.
Detailed Description
The invention is further illustrated by the following examples in conjunction with the accompanying drawings.
Example 1: referring to fig. 1-5, the present embodiment relates to a fluorescence microscope objective with large field of view and high resolution, wherein the magnification of the fluorescence microscope objective is set to be 35 times, the numerical aperture is set to be 0.5, the wavelength visible light range is 505nm to 630nm, the object field of view has a diameter of 10mm, and the object resolution is set to be 0.4 μm. The initial structure of the fluorescence microscope objective adopts a mode of mainly considering the structural characteristics of the photoetching objective; the fluorescence microscope objective is a water immersion objective that achieves a large numerical aperture and provides improved diffraction-limited spatial resolution, which requires the front surface of the first plano-convex lens L1 to be convex or planar to ensure that no bubbles are generated when the objective is immersed in water.
As shown in fig. 1 to 2, the fluorescence microscope objective lens includes a glass slide S, water W, a collimator group G1, a reverse telephoto group G2, a dichroic mirror D, an emission filter E, an achromatic group G3, and a gaussian-like group G4, which are arranged in this order from the object plane to the image plane.
The collimating lens group G1 adopts a plurality of meniscus-shaped thick lenses and positive and negative thin lenses to separately correct the field curvature, and comprises a first plano-convex lens L1, a first convex-concave lens L2, a first concave-convex lens L3 and a second plano-convex lens L4 which are sequentially arranged along the optical axis direction; the structure of the negative telephoto group G2 is basically symmetrical, the aberration is corrected by adopting a positive and negative lens combination method, and the negative telephoto group G2 comprises a first biconcave lens L5, a first biconvex lens L6, a second biconvex lens L7 and a second convex-concave lens L8 which are sequentially arranged along the optical axis direction; the achromatic lens group G3 uses crown glass and flint glass in combination to correct chromatic aberration, and comprises a third biconvex lens L9 and a third convex-concave lens L10 which are sequentially arranged along the optical axis direction; the Gaussian-like lens group G4 adopts a structure form that a positive lens group and a negative lens group are separated and a negative lens group is arranged in front, and comprises a fourth convex-concave lens L11, a second double-concave lens L12, a fifth convex-concave lens L13, a second convex-concave lens L14 and a third convex-concave lens L15 which are sequentially arranged along the optical axis direction, and the fluorescence microscope objective lens adopts a Gaussian-like lens group G4 deformed by the Gaussian lens group to shorten the conjugate distance of an object image.
The principal rays of the incident end of the dichroic mirror D and the emergent end of the emission filter E are strictly parallel, and the lens group of the incident end of the dichroic mirror D and the emergent end of the emission filter E retains aberration to compensate the aberration generated by the two flat plates.
In consideration of energy and stability, a lens material with high transmittance is selected in the design process, the thickness of the lens is reduced while high imaging quality is ensured, and low energy loss of the whole system is ensured; the system considers the high-intensity laser illumination, the cracking of the cemented lens can be caused under the conditions of high-intensity laser illumination and large cemented area, and the cemented lens is avoided during design.
The object plane and the image plane of the existing photoelectric imaging system are both planes, but the design and the processing of a flat-field optical lens with a centimeter-level wide field of view and micron-level resolution are extremely difficult, the field curvature aberration at the edge of the wide field of view is serious, and the optimal imaging position deviates from the image plane, so that the image plane of the fluorescent microscope objective is a curved surface, and the wide field of view and the high resolution of the objective are considered. In addition, the fluorescence microscope objective lens gives consideration to both object space telecentricity and image space telecentricity, the proportional relation of an object image is constant and is determined by the object space telecentricity, and the uniform illumination of an image surface is determined by the image space telecentricity, so that the objective lens has the advantages of small distortion, large depth of field, high resolution and small vignetting.
The imaging quality analysis of the fluorescence microscope objective is shown in fig. 3-5. As shown in fig. 3, the modulation transfer function MTF value of the fluorescence microscope objective lens approaches the diffraction limit in the full field of view of the working band, leaving a large margin for subsequent installation and adjustment; as shown in FIG. 4, the diffuse spot root mean square radius of the fluorescence microscope objective is less than 16 μm over the full field of view of the operating band; as shown in fig. 5, the relative distortion of the fluorescence microscope objective lens is less than 1.6% over the full field of view of the operating band.
Example 2:
referring to fig. 6-10, the present embodiment relates to a fluorescence microscope objective with large field of view and high resolution, wherein the magnification of the fluorescence microscope objective is set to be 10 times, the numerical aperture is set to be 0.3, the wavelength visible light range is 505nm to 630nm, the object field of view has a diameter of 4mm, and the object resolution is set to be 2 μm.
Unlike embodiment 1, the fluorescence microscope objective lens is a dry objective lens, and a lens or a mirror group is placed only at a position between the slide glass S and the dichroic mirror D. As shown in fig. 6 to 7, the fluorescence microscope objective lens includes a glass slide S, air a, spherical lenses L16 to L28, a dichroic mirror D, and an emission filter E, which are arranged in this order from the object plane to the image plane.
Imaging quality analysis of fluorescence microscope objective lens see figures 8-10. As shown in fig. 8, the modulation transfer function MTF value of the fluorescence microscope objective lens approaches the diffraction limit in the full field of view of the working band, leaving a large margin for subsequent installation and adjustment; as shown in FIG. 4, the diffuse spot root mean square radius of the fluorescence microscope objective is less than 22 μm over the full field of view of the operating band; as shown in fig. 5, the relative distortion of the fluorescence microscope objective was less than 0.7% over the full field of view of the operating band.
The above-described embodiments of the present invention should not be construed as limiting the scope of the present invention. Any other corresponding changes and modifications made according to the technical idea of the present invention should be included in the protection scope of the claims of the present invention.
Claims (7)
1. The large-field-of-view high-resolution fluorescence microscope objective is characterized by comprising a glass slide (S), a medium (W), a collimating lens group (G1), an anti-telelens group (G2), a dichroic mirror (D), an emission filter (E), an achromatic lens group (G3) and a Gaussian-like lens group (G4) which are sequentially arranged from an object plane to an image plane; a lens or a lens group is placed on at least one of the position between the glass slide (S) and the dichroic mirror (D), the position between the dichroic mirror (D) and the emission filter (E), and the position behind the emission filter (E); the obtained fluorescence microscope objective has magnification of 10-35 times, numerical aperture of 0.3-0.5, wavelength of 480-680 nm in visible light range, object field diameter of 4-10 mm, and object resolution
(ii) a The anti-telephoto group (G2) includes a first biconcave lens (L5), a first biconvex lens (L6), a second biconvex lens (L7), and a second convex-concave lens (L8) arranged in this order along the optical axis direction; the Gaussian-like lens group comprises a fourth convex-concave lens (L11), a second double-concave lens (L12), a fifth convex-concave lens (L13), a second convex-concave lens (L14) and a third convex-concave lens (L15) which are sequentially arranged along the optical axis direction, and the fluorescence microscope objective lens adopts a Gaussian lensA distorted Gaussian-like lens group (G4) is used to shorten the conjugate distance of the object image.
2. The large-field high-resolution fluorescence microscope objective lens according to claim 1, wherein the collimator lens group (G1) comprises a first plano-convex lens (L1), a first convex-concave lens (L2), a first concave-convex lens (L3) and a second plano-convex lens (L4) which are arranged in sequence along an optical axis direction.
3. The large-field high-resolution fluorescence microscope objective lens according to claim 2, wherein the achromatic lens group (G3) comprises a third biconvex lens (L9) and a third convex-concave lens (L10) arranged in sequence along the optical axis direction.
4. The objective lens of claim 3, wherein the principal rays of the dichroic mirror incident end and the emission filter exit end are strictly parallel, and the aberration of the lens group of the dichroic mirror incident end and the emission filter exit end is retained to compensate the aberration generated by the two flat plates.
5. The large-field-of-view high-resolution fluorescence microscope objective of claim 4, wherein the image plane of the fluorescence microscope objective is a curved plane.
6. The large-field high-resolution fluorescence microscope objective of claim 5, wherein the fluorescence microscope objective has an object-side telecentricity and an image-side telecentricity.
7. The large-field high-resolution fluorescence microscope objective of claim 1, wherein the fluorescence microscope objective initial structure is mainly in the form of a microscope objective structure and has a photoetching objective structural feature.
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| CN111458839B (en) * | 2020-04-21 | 2024-05-31 | 中国科学院西安光学精密机械研究所 | Large-view-field low-distortion lens |
| CN111856737B (en) * | 2020-07-13 | 2021-06-18 | 浙江大学 | Two-photon light field calculation microscope objective |
| CN113900227B (en) * | 2021-10-09 | 2022-07-05 | 中国科学院苏州生物医学工程技术研究所 | Objective lens with large field of view and high resolution broadband |
| CN114967094B (en) * | 2022-04-27 | 2023-06-02 | 华中科技大学 | Telecentric living cell phase contrast microscope lens |
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| CN101031837A (en) * | 2004-07-23 | 2007-09-05 | 通用电气医疗集团尼亚加拉有限公司 | Method and apparatus for fluorescent confocal microscopy |
| CN101292187A (en) * | 2005-10-03 | 2008-10-22 | C.C.M.控股有限公司 | Fluorescence microscope |
| CN206074895U (en) * | 2016-08-30 | 2017-04-05 | 北京大学 | Micro- amplification system |
| CN106842529A (en) * | 2017-01-23 | 2017-06-13 | 清华大学 | Quick three-dimensional micro imaging system |
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| EP3037861A1 (en) * | 2014-12-23 | 2016-06-29 | Commissariat à l'Énergie Atomique et aux Énergies Alternatives | Imaging method, and system, for obtaining an super-resolution image of an object |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101031837A (en) * | 2004-07-23 | 2007-09-05 | 通用电气医疗集团尼亚加拉有限公司 | Method and apparatus for fluorescent confocal microscopy |
| CN101292187A (en) * | 2005-10-03 | 2008-10-22 | C.C.M.控股有限公司 | Fluorescence microscope |
| CN206074895U (en) * | 2016-08-30 | 2017-04-05 | 北京大学 | Micro- amplification system |
| CN106842529A (en) * | 2017-01-23 | 2017-06-13 | 清华大学 | Quick three-dimensional micro imaging system |
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