CN112666696A - Focus scanning method, focus scanning device, and confocal microscope - Google Patents
Focus scanning method, focus scanning device, and confocal microscope Download PDFInfo
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- CN112666696A CN112666696A CN202110063541.9A CN202110063541A CN112666696A CN 112666696 A CN112666696 A CN 112666696A CN 202110063541 A CN202110063541 A CN 202110063541A CN 112666696 A CN112666696 A CN 112666696A
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Abstract
The invention discloses a focal point scanning method, a focal point scanning device and a confocal microscope, wherein the focal point scanning method comprises the following steps: the focal point of the focusing element falls within the immersion medium; changing the refractive index of the immersion medium to adjust the position of the focal point in the axial direction of the focusing element; and detecting imaging information at the focal point using the objective lens and the CCD. The focus scanning method is simple to operate and easy to realize, and the manufacturing cost of the related device can be reduced.
Description
Technical Field
The invention relates to the technical field of precision measurement, in particular to a focus scanning method, a focus scanning device and a confocal microscope.
Background
Point scanning is widely used in confocal microscopy and near-field scanning microscopy. Confocal microscopy was first proposed by m.minsky in the 50 th century, and united states patents were filed in 1957 for stage scanning confocal optical microscopy, in which a point light source was used to illuminate a sample, light carrying sample information was collected by a point detector, and finally a galvanometer was used to scan in the transverse and axial directions to obtain three-dimensional information of the sample. The transverse and longitudinal scanning of the near-field scanning microscope is mainly realized by adopting nano piezoelectric ceramics. The dot scanning structure in the above-described apparatus is high in accuracy, but the structure is precise and complicated, and the manufacturing cost is high.
Disclosure of Invention
The present invention is directed to solving, at least to some extent, one of the technical problems in the related art.
Therefore, the embodiment of the invention provides a focus scanning method, which can realize point scanning by changing the refractive index of the immersed medium, and has simple operation and easy realization.
The embodiment of the invention provides a focus scanning device which is simple in structure, convenient to operate and low in manufacturing cost.
The embodiment of the invention provides a confocal microscope which realizes point scanning by changing the refractive index of an immersed medium, and has the advantages of simple structure and low manufacturing cost.
The focus scanning method according to an embodiment of the present invention includes the steps of:
the focal point of the focusing element falls within the immersion medium;
changing the refractive index of the immersion medium to adjust the position of the focal point in the axial direction of the focusing element; and
the imaging information at the focus is detected by the objective lens and the CCD.
According to the focus scanning method of the embodiment of the present invention, the positions in the axial direction of the focus formed by the focusing elements having different refractive indexes of the immersion medium are different, the refractive index of the immersion medium is continuously changed, and the position in the axial direction of the focus is also continuously changed, so that focus scanning can be realized by the continuous change in the refractive index of the immersion medium.
Therefore, the focus scanning method provided by the embodiment of the invention is simple to operate and easy to realize, so that the manufacturing cost of the related device can be reduced.
In some embodiments, the refractive index of the immersion medium is varied by adjusting the density of the immersion medium.
In some embodiments, the refractive index of the immersion medium is changed by adjusting the temperature of the immersion medium.
In some embodiments, the refractive index of the immersion medium is changed by replacing the immersion medium.
In some embodiments, the refractive index change law is discretized at equal intervals into n ═ nmin+ (m-1) δ, m ═ 1,2,3, …; and completing the change of the refractive index m times, and obtaining the result of step scanning of the focus m times in the axial direction.
The focus scanning apparatus according to an embodiment of the present invention includes: a CCD; a coherent light source that produces a light beam; a collimating lens through which the light beam is collimated; the light beam is collimated and then focused by the focusing element to form a focus; an immersion medium within which the focal point falls so as to adjust a position of the focal point in an axial direction of the focusing element by changing a refractive index of the immersion medium; an objective lens that detects the focal point; and the objective lens detects the focus and focuses and images on the photosensitive surface of the CCD through the focusing lens.
According to the focus scanning device of the embodiment of the invention, the focus formed by the focusing element falls in the immersion medium, and the position of the focus formed by the focusing element in the axial direction is different according to the refractive index of the immersion medium, so that the focus scanning can be realized by continuously changing the refractive index of the immersion medium.
Therefore, the focus scanning device provided by the embodiment of the invention has the advantages of simple structure, convenience in operation and low manufacturing cost.
In some embodiments, the coherent light source, the collimator lens, the focusing element, the immersion medium, the objective lens, the focusing lens, and the CCD are arranged in sequence along an irradiation direction of the light beam.
In some embodiments, the focusing element is a multi-ring band diffractive optical element.
In some embodiments, the immersion medium is disposed adjacent to the focusing element such that the focal point falls within the immersion medium.
The confocal microscope according to the embodiment of the invention comprises the focus scanning device described in any one of the above embodiments.
Drawings
Fig. 1 is a schematic view of a focus scanning apparatus according to an embodiment of the present invention.
Fig. 2 is a linear plot of the refractive index of the immersion medium versus the focal length of the focusing element.
Reference numerals:
a coherent light source 100; a light beam 110;
a collimating lens 200;
a focusing element 300; a focal point 310;
an objective lens 500;
a focusing lens 600;
CCD700。
Detailed Description
Reference will now be made in detail to embodiments of the present invention, examples of which are illustrated in the accompanying drawings. The embodiments described below with reference to the drawings are illustrative and intended to be illustrative of the invention and are not to be construed as limiting the invention.
A focus scanning apparatus of an embodiment of the present invention is described below with reference to the drawings.
As shown in fig. 1, a focus scanning apparatus according to an embodiment of the present invention includes a coherent light source 100, a collimator lens 200, a focusing element 300, an immersion medium 400, an objective lens 500, a focusing lens 600, and a CCD 700. The coherent light source 100, the collimator lens 200, the focusing element 300, the immersion medium 400, the objective lens 500, the focusing lens 600, and the CCD700 are arranged in this order along the irradiation direction of the light beam 110 (the direction from right to left in fig. 1).
As shown in fig. 1, a coherent light source 100 generates a light beam 110, the light beam 110 is collimated by a collimating lens 200, the light beam 110 is collimated and focused by a focusing element 300 to form a focal point 310, and the focal point 310 falls within an immersion medium 400, so that the position of the focal point 310 in the axial direction of the focusing element 300 (the focal length of the focusing element 300) is adjusted by changing the refractive index of the immersion medium 400. The objective lens 500 detects the focal point 310, and then focuses the image on the photosensitive surface of the CCD700 through the focusing lens 600.
It is understood that the refractive index of the immersion medium 400 can be varied by varying the temperature, density, or other parameters of the immersion medium 400. Furthermore, changing the type of immersion medium 400 may also change the refractive index of immersion medium 400, for example immersion medium 400 may be water, air, glycerol, or other medium.
Accordingly, the focus scanning apparatus of the embodiment of the present invention can change the refractive index of the immersion medium 400 by changing at least one of the temperature of the immersion medium 400, the density of the immersion medium 400, and the kind of the immersion medium 400 to achieve a continuous change in the refractive index of the immersion medium 400.
According to the focus scanning apparatus of the embodiment of the present invention, the focus 310 formed by the focusing element 300 falls within the immersion medium 400, and the position of the focus 310 formed by the focusing element 300 in the axial direction is different depending on the refractive index of the immersion medium 400, so that focus scanning can be achieved by continuously changing the refractive index of the immersion medium 400.
Therefore, the focus scanning device provided by the embodiment of the invention has the advantages of simple structure, convenience in operation and low manufacturing cost.
In some embodiments, the focusing element 300 may be a multi-zone diffractive optical element. The multi-zone diffraction optical element is designed by adopting a vector optics diffraction theory and a genetic algorithm, thereby having super-resolution focusing capability. In addition, the multi-ring-band diffractive optical element can be rapidly and optimally designed to achieve the required focusing capacity.
It will be appreciated that the focusing element 300 may also be other optical elements having focusing capabilities.
In some embodiments, as shown in FIG. 1, the immersion medium 400 is positioned adjacent to the focusing element 300 such that the focal point 310 falls within the immersion medium 400. That is, the minimum distance between the immersion medium 400 and the focusing element 300 is greater than or equal to zero. It is understood that the minimum distance between the immersion medium 400 and the focusing element 300 may be zero or greater than zero, provided that the focus point 310 is within the immersion medium 400. Therefore, the focus scanning apparatus of the embodiment of the present invention can adjust the position of the focus 310 in the axial direction by changing the refractive index of the immersion medium 400.
The confocal microscope of the embodiment of the present invention is described below.
The confocal microscope according to the embodiment of the invention comprises the focus scanning device described in any one of the above embodiments. Other configurations of confocal microscopes are well known to those skilled in the art and will not be described.
A focus scanning method of an embodiment of the present invention is described below with reference to the drawings.
As shown in fig. 1, a focus scanning method according to an embodiment of the present invention includes the steps of:
the focal point 310 of the focusing element 300 falls within the immersion medium 400;
changing the refractive index of the immersion medium 400 to adjust the position of the focal point 310 in the axial direction of the focusing element 300; and
imaging information at the focal point 310 is detected using the objective lens 500 and the CCD 700.
According to the focus scanning method of the embodiment of the present invention, the position of the focus 310 formed by the focusing elements 300 having different refractive indexes of the immersion medium 400 in the axial direction (the focal length of the focusing elements 300) is different, the refractive index of the immersion medium 400 is continuously changed, and the position of the focus 310 in the axial direction is also continuously changed, so that the focus scanning can be realized by the continuous change of the refractive index of the immersion medium 400.
Therefore, the focus scanning method provided by the embodiment of the invention is simple to operate and easy to realize, so that the manufacturing cost of the related device can be reduced.
In some embodiments, the focus scanning method of embodiments of the present invention changes the refractive index of the immersion medium 400 by adjusting the density of the immersion medium 400, thereby changing the position of the focus 310 in the axial direction.
In some embodiments, the focus scanning method of the embodiments of the present invention changes the refractive index of the immersion medium 400 by adjusting the temperature of the immersion medium 400, thereby changing the position of the focus 310 in the axial direction.
In some embodiments, the focus scanning method of embodiments of the present invention changes the refractive index of the immersion medium 400 by replacing the immersion medium 400, thereby changing the position of the focus 310 in the axial direction.
It is understood that the focus scanning method of the present invention can employ the method of at least one of the above three embodiments to change the refractive index of the immersion medium 400 to achieve a continuous change in the refractive index of the immersion medium 400, thereby achieving a continuous change in the position of the focus 310 in the axial direction.
In some embodiments, the refractive index change law is discretized at equal intervals into n ═ nmin+ (m-1) δ, m ═ 1,2,3, …; the m refractive index changes are completed, and the results of the axial step scanning of the m focal points 310 are obtained. That is, the position of the focal point 310 in the axial direction (the focal length of the focusing element 300) that is actually measured corresponds to the refractive index one-to-one. The refractive index is discretely sampled at equal intervals, and m values are extracted.
As shown in fig. 2, the position of the focal point 310 in the axial direction (focal length of the focusing element 300) measured in practice is plotted in a linear relationship with the refractive index. As can be seen from fig. 2, the position of the focal point 310 in the axial direction (the focal length of the focusing element 300) has a good linear relationship with the refractive index.
Therefore, the focus scanning method of the embodiment of the invention can perform focus scanning by using the change of the refractive index, is simple to operate and easy to realize, and can reduce the manufacturing cost of the focus scanning device for realizing the focus scanning method.
In the description of the present invention, it is to be understood that the terms "central," "longitudinal," "lateral," "length," "width," "thickness," "upper," "lower," "front," "rear," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," "outer," "clockwise," "counterclockwise," "axial," "radial," "circumferential," and the like are used in the orientations and positional relationships indicated in the drawings for convenience in describing the invention and to simplify the description, and are not intended to indicate or imply that the referenced devices or elements must have a particular orientation, be constructed and operated in a particular orientation, and are therefore not to be considered limiting of the invention.
Furthermore, the terms "first", "second" and "first" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include at least one such feature. In the description of the present invention, "a plurality" means at least two, e.g., two, three, etc., unless specifically limited otherwise.
In the present invention, unless otherwise expressly stated or limited, the terms "mounted," "connected," "secured," and the like are to be construed broadly and can, for example, be fixedly connected, detachably connected, or integrally formed; may be mechanically coupled, may be electrically coupled or may be in communication with each other; they may be directly connected or indirectly connected through intervening media, or they may be connected internally or in any other suitable relationship, unless expressly stated otherwise. The specific meanings of the above terms in the present invention can be understood by those skilled in the art according to specific situations.
In the present invention, unless otherwise expressly stated or limited, the first feature "on" or "under" the second feature may be directly contacting the first and second features or indirectly contacting the first and second features through an intermediate. Also, a first feature "on," "over," and "above" a second feature may be directly or diagonally above the second feature, or may simply indicate that the first feature is at a higher level than the second feature. A first feature being "under," "below," and "beneath" a second feature may be directly under or obliquely under the first feature, or may simply mean that the first feature is at a lesser elevation than the second feature.
In the present disclosure, the terms "one embodiment," "some embodiments," "an example," "a specific example," or "some examples" and the like mean that a specific feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the present disclosure. In this specification, the schematic representations of the terms used above are not necessarily intended to refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Furthermore, various embodiments or examples and features of different embodiments or examples described in this specification can be combined and combined by one skilled in the art without contradiction.
Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention, and that variations, modifications, substitutions and alterations can be made to the above embodiments by those of ordinary skill in the art within the scope of the present invention.
Claims (10)
1. A focus scanning method, comprising the steps of:
the focal point of the focusing element falls within the immersion medium;
changing the refractive index of the immersion medium to adjust the position of the focal point in the axial direction of the focusing element; and
the imaging information at the focus is detected by the objective lens and the CCD.
2. The focus scanning method according to claim 1, wherein a refractive index of the immersion medium is changed by adjusting a density of the immersion medium.
3. The focus scanning method according to claim 1, wherein a refractive index of the immersion medium is changed by adjusting a temperature of the immersion medium.
4. The focus scanning method according to claim 1, wherein the refractive index of the immersion medium is changed by replacing the immersion medium.
5. The focus scanning method according to claim 1, wherein the refractive index change law is dispersed at equal intervals as n-nmin+ (m-1) δ, m ═ 1,2,3, …; and completing the change of the refractive index m times, and obtaining the result of step scanning of the focus m times in the axial direction.
6. A focus scanning apparatus, comprising:
CCD；
a coherent light source that produces a light beam;
a collimating lens through which the light beam is collimated;
the light beam is collimated and then focused by the focusing element to form a focus;
an immersion medium within which the focal point falls so as to adjust a position of the focal point in an axial direction of the focusing element by changing a refractive index of the immersion medium;
an objective lens that detects the focal point; and
and the objective lens detects the focus and focuses and images on the photosensitive surface of the CCD through the focusing lens.
7. The focus scanning apparatus according to claim 6, wherein said coherent light source, said collimator lens, said focusing element, said immersion medium, said objective lens, said focusing lens, and said CCD are disposed in order in an irradiation direction of said light beam.
8. The focal scanning device of claim 6, wherein the focusing element is a multi-ring diffractive optical element.
9. The focal scanning device of claim 7, wherein the immersion medium is disposed adjacent to the focusing element such that the focal point falls within the immersion medium.
10. A confocal microscope comprising the focus scanning apparatus of any one of claims 6 to 9.
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| CN202110063541.9A CN112666696A (en) | 2021-01-18 | 2021-01-18 | Focus scanning method, focus scanning device, and confocal microscope |
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| CN1906520A (en) * | 2004-02-27 | 2007-01-31 | 浜松光子学株式会社 | Microscope and sample observation method |
| US20080106711A1 (en) * | 2004-11-18 | 2008-05-08 | Carl Zeiss Smt Ag | Projection Lens System of a Microlithographic Projection Exposure Installation |
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Application publication date: 20210416 |