CN115508994A - Microscopic imaging system - Google Patents
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- CN115508994A CN115508994ACN202211115900.1ACN202211115900ACN115508994ACN 115508994 ACN115508994 ACN 115508994ACN 202211115900 ACN202211115900 ACN 202211115900ACN 115508994 ACN115508994 ACN 115508994A
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- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
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- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N2021/6463—Optics
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- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
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Abstract
Description
Translated fromChinese技术领域technical field
本发明涉及成像系统技术领域，特别是涉及一种显微成像系统。The invention relates to the technical field of imaging systems, in particular to a microscopic imaging system.
背景技术Background technique
荧光显微镜是利用滤光器件将光源发出光形成特定波长光，激发样品内的荧光物质发射出荧光，进而在显微镜下观察样品的形态或者结构。其主要应用于生物领域，用于观察和分辨样品中某些化学成分或者细胞组分情况，可以用于细胞结构或者生物化学成分等研究。Fluorescence microscopes use filters to convert the light emitted by the light source into light of a specific wavelength, which excites the fluorescent substances in the sample to emit fluorescence, and then observes the shape or structure of the sample under the microscope. It is mainly used in the biological field to observe and distinguish certain chemical components or cell components in samples, and can be used for research on cell structure or biochemical components.
现有的荧光显微镜是在复式显微镜的架构上安装荧光装置集合而成，荧光装置产生出特定波长的激发光。现有的荧光显微成像系统采用背照式照明，即照明模块与成像模块分别在样品两侧，不能用于非透明样品的观测。Existing fluorescence microscopes are formed by installing fluorescence devices on the structure of a compound microscope, and the fluorescence devices generate excitation light of a specific wavelength. Existing fluorescence microscopy imaging systems use back-illuminated illumination, that is, the illumination module and the imaging module are located on both sides of the sample, which cannot be used for the observation of non-transparent samples.
发明内容Contents of the invention
本发明的目的是提供一种显微成像系统，能够用于观测非透明样品，并且能够提高照射到样品上光的均匀性。The purpose of the present invention is to provide a microscopic imaging system, which can be used to observe non-transparent samples, and can improve the uniformity of light irradiated on the samples.
为实现上述目的，本发明提供如下技术方案：To achieve the above object, the present invention provides the following technical solutions:
一种显微成像系统，包括照明装置、聚光装置、第一透镜组、第二透镜组、第一光学元件和成像面，所述照明装置用于发出照射待测样的光，所述聚光装置用于将所述照明装置的出射光成像到所述第一透镜组的入瞳处，所述第一透镜组用于将所述照明装置出射光的一次像成像到无穷远处，以照射到所述待测样；A microscopic imaging system, comprising an illuminating device, a light concentrating device, a first lens group, a second lens group, a first optical element and an imaging surface, the illuminating device is used to emit light for illuminating a sample to be measured, and the concentrating The light device is used to image the outgoing light of the lighting device to the entrance pupil of the first lens group, and the first lens group is used to image the primary image of the outgoing light of the lighting device to infinity, so as to irradiating the sample to be tested;
所述第一光学元件设置于所述第一透镜组和所述第二透镜组之间，用于使所述待测样发出的、通过所述第一透镜组后的光与由所述聚光装置入射到所述第一透镜组的光分离，所述第二透镜组用于将所述待测样发出的、通过所述第一透镜组后的光成像到所述成像面。The first optical element is arranged between the first lens group and the second lens group, and is used to combine the light emitted by the sample to be measured and passed through the first lens group with the light collected by the condenser. The optical device separates the light incident to the first lens group, and the second lens group is used to image the light emitted by the sample to be measured and passed through the first lens group to the imaging plane.
优选的，所述第一光学元件用于将所述聚光装置出射光反射到所述第一透镜组，以及将所述待测样发出的、通过所述第一透镜组后的光透射到所述第二透镜组；Preferably, the first optical element is used to reflect the light emitted by the light concentrating device to the first lens group, and transmit the light emitted by the sample to be measured and passed through the first lens group to the first lens group. the second lens group;
或者，所述第一光学元件用于将所述聚光装置出射光透射到所述第一透镜组，以及将所述待测样发出的、通过所述第一透镜组后的光反射到所述第二透镜组。Alternatively, the first optical element is used to transmit the light emitted by the light concentrating device to the first lens group, and reflect the light emitted by the sample to be measured and passed through the first lens group to the first lens group. Describe the second lens group.
优选的，还包括设置在所述第二透镜组靠近所述第一透镜组一侧的第一滤光元件，所述第一滤光元件用于允许所述待测样发出光透过，阻止所述照明装置出射光透过；Preferably, it also includes a first filter element arranged on the side of the second lens group close to the first lens group, the first filter element is used to allow the light emitted by the sample to be tested to pass through, and prevent The light emitted by the lighting device passes through;
和/或，还包括设置在所述第二透镜组远离所述第一透镜组一侧的第二滤光元件，所述第二滤光元件用于允许所述待测样发出光透过，阻止所述照明装置出射光透过。And/or, it also includes a second filter element arranged on the side of the second lens group away from the first lens group, the second filter element is used to allow the light emitted by the sample to be measured to pass through, preventing the light emitted by the illuminating device from passing through.
优选的，所述照明装置包括第三滤光元件，所述第三滤光元件用于允许光源发出光中符合要求的波段光透过，而阻止其它波段光透过。Preferably, the illuminating device includes a third filter element, and the third filter element is used to allow the light in the wavelength band that meets the requirements in the light emitted by the light source to pass through, while preventing the light in other wavelength bands from passing through.
优选的，所述照明装置还包括分别发出不同波段光的第一光源和第二光源，所述第三滤光元件为双带通滤光元件。Preferably, the illuminating device further includes a first light source and a second light source respectively emitting light in different wavelength bands, and the third filter element is a dual-bandpass filter element.
优选的，所述照明装置包括光源、准直元件和光阑，所述准直元件用于将所述光源发出光进行准直，准直光通过所述光阑发射出。Preferably, the illuminating device includes a light source, a collimating element and an aperture, the collimating element is used to collimate the light emitted by the light source, and the collimated light is emitted through the aperture.
优选的，还包括对焦装置，用于测量所述第一透镜组到所述待测样的距离，并根据得到的距离控制所述第一透镜组移动，使所述待测样处于所述第一透镜组的焦面上。Preferably, it also includes a focusing device, which is used to measure the distance between the first lens group and the sample to be measured, and control the movement of the first lens group according to the obtained distance, so that the sample to be measured is in the first position. The focal plane of a lens group.
优选的，所述对焦装置具体用于根据得到的距离控制所述第一透镜组沿着自身光轴方向移动，使所述待测样处于所述第一透镜组的焦面上。Preferably, the focusing device is specifically used to control the movement of the first lens group along its own optical axis according to the obtained distance, so that the sample to be measured is on the focal plane of the first lens group.
优选的，所述第一透镜组的焦距小于所述第二透镜组的焦距，所述第二透镜组的焦距小于等于100毫米。Preferably, the focal length of the first lens group is shorter than the focal length of the second lens group, and the focal length of the second lens group is less than or equal to 100 millimeters.
优选的，所述聚光装置包括单透镜、胶合透镜、球面透镜、非球面透镜中的任意一个或者任意多个的组合。Preferably, the light concentrating device includes any one or a combination of single lenses, cemented lenses, spherical lenses, and aspheric lenses.
由上述技术方案可知，本发明所提供的显微成像系统，照明装置用于发出照射待测样的光，聚光装置用于将照明装置的出射光成像到第一透镜组的入瞳处，第一透镜组用于将照明装置出射光的一次像成像到无穷远处，以照射到待测样。第一光学元件设置于第一透镜组和第二透镜组之间，使待测样发出的、通过第一透镜组后的光与由聚光装置入射到第一透镜组的光分离，第二透镜组将待测样发出的、通过第一透镜组后的光成像到成像面，实现对待测样的光学成像。It can be seen from the above technical solution that in the microscopic imaging system provided by the present invention, the illuminating device is used to emit light for illuminating the sample to be tested, and the concentrating device is used to image the outgoing light of the illuminating device to the entrance pupil of the first lens group, The first lens group is used to image the primary image of the light emitted by the illuminating device to infinity so as to irradiate the sample to be tested. The first optical element is arranged between the first lens group and the second lens group, so that the light emitted by the sample to be measured and passed through the first lens group is separated from the light incident on the first lens group by the light-collecting device, and the second The lens group images the light emitted by the sample to be measured and passed through the first lens group to the imaging surface, so as to realize the optical imaging of the sample to be measured.
本发明显微成像系统能够用于观测非透明样品，并且通过聚光装置将照明装置的出射光成像到第一透镜组的入瞳处，通过第一透镜组将光的一次像成像到无穷远处而照射到待测样，能够提高照射到样品上光的均匀性。The microscopic imaging system of the present invention can be used to observe non-transparent samples, and image the outgoing light of the illuminating device to the entrance pupil of the first lens group through the light collecting device, and image the primary image of the light to infinity through the first lens group Wherever it irradiates the sample to be tested, the uniformity of the light irradiated on the sample can be improved.
附图说明Description of drawings
为了更清楚地说明本发明实施例或现有技术中的技术方案，下面将对实施例或现有技术描述中所需要使用的附图作简单地介绍，显而易见地，下面描述中的附图仅仅是本发明的一些实施例，对于本领域普通技术人员来讲，在不付出创造性劳动的前提下，还可以根据这些附图获得其他的附图。In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings that need to be used in the description of the embodiments or the prior art. Obviously, the accompanying drawings in the following description are only These are some embodiments of the present invention. Those skilled in the art can also obtain other drawings based on these drawings without creative work.
图1为本发明实施例提供的一种显微成像系统的示意图；FIG. 1 is a schematic diagram of a microscopic imaging system provided by an embodiment of the present invention;
图2为本发明又一实施例提供的一种显微成像系统的示意图；2 is a schematic diagram of a microscopic imaging system provided by another embodiment of the present invention;
图3为本发明又一实施例提供的一种显微成像系统的示意图；3 is a schematic diagram of a microscopic imaging system provided by another embodiment of the present invention;
图4为本发明又一实施例提供的一种显微成像系统的示意图。Fig. 4 is a schematic diagram of a microscopic imaging system provided by another embodiment of the present invention.
具体实施方式detailed description
为了使本技术领域的人员更好地理解本发明中的技术方案，下面将结合本发明实施例中的附图，对本发明实施例中的技术方案进行清楚、完整地描述，显然，所描述的实施例仅仅是本发明一部分实施例，而不是全部的实施例。基于本发明中的实施例，本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例，都应当属于本发明保护的范围。In order to enable those skilled in the art to better understand the technical solutions in the present invention, 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 The embodiments are only some of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without making creative efforts shall fall within the protection scope of the present invention.
请参考图1，图1为本实施例提供的一种显微成像系统的示意图，如图所示，所述显微成像系统包括照明装置10、聚光装置11、第一透镜组12、第二透镜组13、第一光学元件15和成像面14，所述照明装置10用于发出照射待测样25的光，所述聚光装置11用于将所述照明装置10的出射光成像到所述第一透镜组12的入瞳处，所述第一透镜组12用于将所述照明装置10出射光的一次像成像到无穷远处，以照射到所述待测样25。Please refer to FIG. 1. FIG. 1 is a schematic diagram of a microscopic imaging system provided in this embodiment. Two
所述第一光学元件15设置于所述第一透镜组12和所述第二透镜组13之间，用于使所述待测样发出的、通过所述第一透镜组12后的光与由所述聚光装置11入射到所述第一透镜组12的光分离，所述第二透镜组13用于将所述待测样25发出的、通过所述第一透镜组12后的光成像到所述成像面14。The first
照明装置10的出射光依次通过聚光装置11、第一透镜组12照射到待测样25，使待测样25发出光，待测样25发出的光通过第一透镜组12后，通过第二透镜组13成像到成像面14。从而实现对待测样的光学成像。The light emitted by the
通过第一光学元件15使待测样25发出的、通过第一透镜组12后的光，与由聚光装置11入射到第一透镜组12的光分离，使待测样25发出的、通过第一透镜组12后的光进入第二透镜组13。Through the first
本实施例的显微成像系统能够用于观测透明样品或者非透明样品，并且通过聚光装置将照明装置的出射光成像到第一透镜组的入瞳处，通过第一透镜组将光的一次像成像到无穷远处而照射到待测样，能够使照射到待测样的光更均匀，能够提高照射到样品上光的均匀性。The microscopic imaging system of this embodiment can be used to observe transparent samples or non-transparent samples, and the outgoing light of the illuminating device is imaged to the entrance pupil of the first lens group through the light collecting device, and the primary lens of the light is captured by the first lens group Imaging to infinity and irradiating the sample to be tested can make the light irradiated to the sample to be tested more uniform, and can improve the uniformity of light irradiated on the sample.
聚光装置11将照明装置10的出射光进行汇聚，成像到第一透镜组12的入瞳处。可选的，聚光装置11包括单透镜、胶合透镜、球面透镜、非球面透镜中的任意一个或者任意多个的组合。聚光装置11可包括一个或者多个单透镜，也可包含二向色镜或者棱镜。The
可选的，第一透镜组12包括单透镜、胶合透镜、球面透镜、非球面透镜中的任意一个或者任意多个的组合。第二透镜组13包括单透镜、胶合透镜、球面透镜、非球面透镜中的任意一个或者任意多个的组合。Optionally, the
可选的，第一光学元件可以用于将所述聚光装置出射光反射到所述第一透镜组，以及将所述待测样发出的、通过所述第一透镜组后的光透射到所述第二透镜组。可参考图1所示，聚光装置11出射光入射到第一光学元件15，被反射到第一透镜组12。待测样25的发出光通过第一透镜组12后，透射过第一光学元件15而后进入第二透镜组13。Optionally, the first optical element can be used to reflect the light emitted by the light concentrating device to the first lens group, and transmit the light emitted by the sample to be measured and passed through the first lens group to the first lens group. the second lens group. Referring to FIG. 1 , light emitted from the
可选的，第一光学元件可以是将所述聚光装置出射光透射到所述第一透镜组，以及将所述待测样发出的、通过所述第一透镜组后的光反射到所述第二透镜组。请参考图2，图2为又一实施例提供的一种显微成像系统的示意图，如图所示，聚光装置11出射光入射到第一光学元件16，透射后进入到第一透镜组12。待测样25的发出光通过第一透镜组12后，被第一光学元件16反射而进入第二透镜组13。Optionally, the first optical element can transmit the light emitted by the light concentrating device to the first lens group, and reflect the light emitted by the sample to be measured and passed through the first lens group to the first lens group. Describe the second lens group. Please refer to FIG. 2, which is a schematic diagram of a microscopic imaging system provided by another embodiment. As shown in the figure, the light emitted by the
可选的，第一光学元件可采用但不限于二向色镜或者棱镜。Optionally, the first optical element may be, but not limited to, a dichroic mirror or a prism.
优选的，所述显微成像系统还包括设置在所述第二透镜组靠近所述第一透镜组一侧的第一滤光元件，所述第一滤光元件用于允许所述待测样发出光透过，阻止所述照明装置出射的激发光透过；和/或，还包括设置在所述第二透镜组远离所述第一透镜组一侧的第二滤光元件，所述第二滤光元件用于允许所述待测样发出光透过，阻止所述照明装置出射的激发光透过。通过第一滤光元件或者第二滤光元件，只允许待测样发出光通过第二透镜组进行成像，阻挡激发光入射到成像面而干扰成像结果，有助于提高对样品的光学成像效果。请参考图3，图3为又一实施例提供的一种显微成像系统的示意图，如图所示，在第二透镜组13和第一光学元件15之间设置有第一滤光元件17，在第二透镜组13和成像面14之间设置有第二滤光元件18。Preferably, the microscopic imaging system further includes a first filter element arranged on the side of the second lens group close to the first lens group, the first filter element is used to allow the sample to be tested transmit the emitted light, prevent the excitation light emitted by the illuminating device from passing through; and/or, further include a second filter element arranged on the side of the second lens group away from the first lens group, the first The second filter element is used to allow the light emitted by the sample to pass through, and prevent the excitation light emitted by the illuminating device from passing through. Through the first filter element or the second filter element, only the light emitted by the sample to be tested is allowed to pass through the second lens group for imaging, preventing the excitation light from entering the imaging surface and interfering with the imaging result, which helps to improve the optical imaging effect of the sample . Please refer to FIG. 3 . FIG. 3 is a schematic diagram of a microscopic imaging system provided by another embodiment. As shown in the figure, a
优选的，照明装置可包括第三滤光元件，所述第三滤光元件用于允许光源发出光中符合要求的波段光透过，而阻止其它波段光透过。在照明装置中通过设置第三滤光元件，从光源的发出光中过滤出符合要求的波段光作为激发光，并且通过第三滤光元件阻止光源中与待测样品荧光相同的波段通过，保证待测样品荧光中不包含光源引入的杂散光，有助于提高对样品的光学成像效果。Preferably, the illuminating device may include a third filter element, the third filter element is used to allow the light in the required wavelength bands of the light emitted by the light source to pass through, while preventing the light in other wavelength bands from passing through. In the lighting device, by setting the third filter element, the light of the required wavelength band is filtered out from the light emitted by the light source as the excitation light, and the third filter element prevents the passage of the same wavelength band in the light source as the fluorescent light of the sample to be measured to ensure The fluorescence of the sample to be tested does not contain the stray light introduced by the light source, which helps to improve the optical imaging effect of the sample.
进一步可选的，照明装置可包括光源、准直元件和光阑，所述准直元件用于将所述光源发出光进行准直，准直光通过所述光阑发射出。通过光阑限定照明装置出射光的视场范围，限定照射到待测样的激发光的视场范围。Further optionally, the illuminating device may include a light source, a collimating element and an aperture, the collimating element is used to collimate the light emitted by the light source, and the collimated light is emitted through the aperture. The field of view range of the light emitted by the illumination device is limited by the aperture, and the field of view range of the excitation light irradiated to the sample to be measured is limited.
可参考图3所示，照明装置10包括光源20、准直元件21、光阑22和第三滤光元件19。可选的，光源20可采用但不限于LED光源或者LD光源。准直元件21可包括一个或者多个透镜，包括但不限于单透镜或者胶合透镜，可采用球面透镜或者非球面透镜。光阑22可采用但不限于矩形光阑或者圆形光阑。Referring to FIG. 3 , the illuminating
优选的可参考图4所示，图4为又一实施例提供的一种显微成像系统的示意图，如图所示照明装置可包括分别发出不同波段光的第一光源100和第二光源103，在实际应用中可根据待测样25的检测需求选择第一光源100或者第二光源103的出射光波段范围。若待测样25的激发波段对应于第一光源100则开启第一光源100，若待测样的激发波段对应于第二光源103则开启第二光源103。Preferably, as shown in FIG. 4, FIG. 4 is a schematic diagram of a microscopic imaging system provided by another embodiment. As shown in the figure, the illuminating device may include a first
可选的如图4所示，第一光源100发出光通过第一准直元件101后形成准直光，通过光阑102后入射到第二光学元件106，从第二光学元件106透射出。第二光源103发出光通过第二准直元件104后形成准直光，通过光阑105后入射到第二光学元件106，从第二光学元件106反射出。第二光学元件106可采用但不限于二向色镜、棱镜等。Optionally, as shown in FIG. 4 , light emitted from the first
优选的，第三滤光元件19为双带通滤光元件，使得本显微成像系统使用一组滤光元件通过分时成像，能够实现两种不同波段荧光的观测。第三滤光元件19后边可以只放置一个成像面14，配合双带通滤光元件，可以完成对双色荧光探测，只设置一个成像面14可以减小系统体积，减少双系统装配匹配难度，降低成本；通过对两个光源分时操作(即第一光源100和第二光源103发出的光波长不同，在使用的时候轮流开启)，成像面14可以通过时间来判断具体的荧光波长，同时一次只开启一个光源，会降低样品处的光功率，也有利于保护待测物荧光寿命。Preferably, the
进一步优选的，所述显微成像系统还可包括对焦装置，用于控制所述第一透镜组12移动使所述待测样25处于所述第一透镜组12的焦面上。优选的，对焦装置可用于测量所述第一透镜组12到所述待测样25的距离，并根据得到的距离控制所述第一透镜组12移动，使所述待测样25处于所述第一透镜组12的焦面上。可通过与第一透镜组12连接的驱动电机，驱动第一透镜组12移动。具体的，对焦装置可具体用于根据得到的距离控制所述第一透镜组12沿着自身光轴方向移动，使所述待测样25处于所述第一透镜组12的焦面上。Further preferably, the microscopic imaging system may further include a focusing device for controlling the movement of the
优选的，本实施例的显微成像系统中第一透镜组12的焦距小于所述第二透镜组13的焦距，所述第二透镜组13的焦距小于等于100毫米。本显微成像系统采用了短焦镜头(焦距40-100mm)作为第二透镜组13，成像面可采用CMOS/CCD/SCMOS等成像芯片，像元尺寸为1-10um，优选为2-4um，大大减小了第二透镜组13和相机的体积，使得系统体积更加紧凑。Preferably, in the microscopic imaging system of this embodiment, the focal length of the
以上对本发明所提供的一种显微成像系统进行了详细介绍。本文中应用了具体个例对本发明的原理及实施方式进行了阐述，以上实施例的说明只是用于帮助理解本发明的方法及其核心思想。应当指出，对于本技术领域的普通技术人员来说，在不脱离本发明原理的前提下，还可以对本发明进行若干改进和修饰，这些改进和修饰也落入本发明权利要求的保护范围内。A microscopic imaging system provided by the present invention has been introduced in detail above. In this paper, specific examples are used to illustrate the principle and implementation of the present invention, and the descriptions of the above embodiments are only used to help understand the method and core idea of the present invention. It should be pointed out that for those skilled in the art, without departing from the principles of the present invention, some improvements and modifications can be made to the present invention, and these improvements and modifications also fall within the protection scope of the claims of the present invention.
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