WO2021129588A1 - Fluorescent lighting device and microscopy imaging system - Google Patents

Abstract

A fluorescent lighting device (00) and a microscopy imaging system (20). The fluorescent lighting device (00) comprises a cubic light-filtering body (10). The cubic light-filtering body (10) comprises: a housing (110) having a first opening (111) and a second opening (112) arranged opposite to each other; a multi-color fluorescence light source (120) fixed to the interior of the housing (110); a first multiband filter (130) provided at a light emitting side of the multi-color fluorescence light source (120); and a multiband dichroic mirror (140) provided along a first optical axis (X) at one side of the first multiband filter (130) away from the multi-color fluorescence light source (120), wherein light is propagated along a second optical axis (Y) after reflected by the multiband dichroic mirror (140), the second optical axis (Y) passes through the first opening (111) and the second opening (112), and the second optical axis (Y) intersects with the first optical axis (X).

Description

Fluorescence lighting device and microscopic imaging system

This application claims the priority of the Chinese patent application filed with the Chinese Patent Office with the application number 201911358186.7 on December 25, 2019, and the entire content of the application is incorporated into this application by reference.

Technical field

The embodiments of the present application relate to the technical field of fluorescence microscopy imaging, for example, to a fluorescence illumination device and a microscopy imaging system.

Background technique

With the development of microscopic imaging technology, the fluorescence microscope, as a device for experimental observation or testing, has become more and more widely used. Generally, in order to realize the observation of different fluorescent substances in the sample, a variety of different fluorescent light sources need to be equipped. The fluorescent light source can use mercury and xenon arc lamps, but their life is short and the cost is high; or a light emitting diode (LED) light source can be used as the fluorescent light source, and the LED light source has a better light output effect and lower cost. However, when a variety of different LED light sources are required to provide fluorescence, the problem of light source switching needs to be considered.

The related technology provides a light cube (also called a filter cube), which integrates a monochromatic light source, a monochromatic filter and a monochromatic dichroic mirror in a filter cube, and sets multiple If you have a light cube, you can switch between different light cubes as needed to provide a variety of light sources with different wavelengths. However, in this fluorescence microscope structure, each light cube can only provide one light source; at the same time, each light cube must be equipped with its own set of independent monochromatic filters and monochromatic dichroic mirrors. As a result, not only makes the overall structure of the fluorescence microscope more complicated, but also its cost is higher.

Summary of the invention

The embodiments of the present application provide a fluorescent lighting device and a microscopic imaging system, so as to simplify the structure of the fluorescent lighting device while providing a variety of fluorescent light sources of different wavelengths, thereby simplifying the structure of the microscopic imaging system and reducing the fluorescent lighting. The cost of the device and the microscopic imaging system.

An embodiment of the application provides a fluorescent lighting device, the fluorescent lighting device includes: a filter cube, and the filter cube includes:

A housing, the housing having a first opening and a second opening, the first opening and the second opening are arranged opposite to each other;

Multicolor fluorescent light source, fixed inside the housing;

The first multi-band filter is arranged on the light-emitting side of the multi-color fluorescent light source;

A multi-band dichroic mirror is arranged on the side of the first multi-band filter away from the multi-color fluorescent light source along the first optical axis;

After being reflected by the multi-band dichroic mirror, the light propagates along a second optical axis; the second optical axis passes through the first opening and the second opening, and the second optical axis and the first One optical axis crosses.

The embodiment of the present application also provides a microscopic imaging system. The microscopic imaging system includes any of the above-mentioned fluorescent lighting devices; and further includes: a sample stage, an objective lens, and an image sensor;

The sample stage, the objective lens, the multi-band dichroic mirror, and the image sensor are sequentially arranged along the second optical axis.

Description of the drawings

FIG. 1 is a schematic structural diagram of a fluorescent lighting device provided by an embodiment of the present application;

2 is a schematic diagram of the structure of a multi-color LED light source in a fluorescent lighting device provided by an embodiment of the present application;

Fig. 3 is a schematic structural diagram of another fluorescent lighting device provided by an embodiment of the present application;

4 is a schematic structural diagram of yet another fluorescent lighting device provided by an embodiment of the present application;

Fig. 5 is a schematic structural diagram of a microscopic imaging system provided by an embodiment of the present application.

Detailed ways

The fluorescent lighting device (also referred to as "fluorescence light source" or "microscopic light source") and microscopic imaging system (also referred to as "fluorescence microscope" or "microscope structure") provided by the embodiments of the present application proposes a The new type of multi-channel microscopy light source scheme is exemplary: using a multi-band filter and a multi-band dichroic mirror, a variety of different wavelengths of fluorescent light sources are integrated into the same filter cube. Therefore, while providing a variety of fluorescent light sources with different wavelength band requirements, switching between multiple different filter cubes can be avoided; further, the structure of the fluorescent light source and the fluorescent microscope is simplified, and the cost is saved.

It should be noted that the filter cube is only a commonly used name in this field, but the present application does not limit the shape of the filter cube to "cube".

Hereinafter, with reference to FIGS. 1 to 5, the fluorescent lighting device and the microscopic imaging system provided in the embodiments of the present application will be exemplarily described.

Fig. 1 is a schematic structural diagram of a fluorescent lighting device provided by an embodiment of the present application. 1, the fluorescent lighting device 00 includes a filter cube 10 (also referred to as "light cube 10"), the filter cube 10 includes: a housing 110, the housing 110 has a first opening 111 and a second opening 112, the first The opening 111 and the second opening 112 are arranged oppositely; the multi-color fluorescent light source 120 is fixed inside the housing 110; the first multi-wavelength filter 130 is arranged on the light exit side of the multi-color fluorescent light source 120; the multi-wavelength dichroic mirror 140, Along the first optical axis X, it is arranged on the side of the first multi-band filter 130 away from the multi-color fluorescent light source 120; after being reflected by the multi-band dichroic mirror 140, the light propagates along the second optical axis Y; The axis Y passes through the first opening 111 and the second opening 112, and the second optical axis Y crosses the first optical axis X. Wherein, the first opening 111 and the second opening 112 are arranged oppositely, which means that the projections of the first opening 111 and the second opening 112 on a plane perpendicular to the second optical axis Y at least partially overlap.

Wherein, along the first optical axis X, the light emitted by the multi-color fluorescent light source 120 is filtered by the first multi-band filter 130 to form a specific wavelength of fluorescence, and then the specific wavelength of fluorescence is reflected by the multi-band dichroic mirror 140 , Shoot out through the first opening 111. The emitted fluorescence irradiates the sample to excite the sample to emit light; along the second optical axis Y, the light emitted by the sample successively passes through the first opening 111, the multi-band dichroic mirror 140, and the second opening 112 to be detected by the photoelectric in the microscope structure The device receives.

Among them, the multi-color fluorescent light source 120 can emit light of multiple different wavebands, the first multi-wavelength filter 130 can filter light of multiple wavebands, and the multi-wavelength dichroic mirror 140 can correspondingly treat light of multiple wavebands. Reflect, so as to use the same light cube to provide a variety of fluorescence in different wavelength bands.

The multi-band dichroic mirror 140 includes an incident surface and a light-transmitting surface. The incident surface and the light-transmitting surface are two surfaces of the multi-band dichroic mirror 140 opposite to each other. The incident surface faces the side of the multi-band dichroic mirror 140. On the incident side, the light-transmitting surface faces the light-transmitting side of the multi-band dichroic mirror 140. The incident surface can reflect the light emitted by the first multi-band filter 130 and transmit the light excited by the sample. The incident surface of the multi-band dichroic mirror 140 is set to face the first opening 111, and the angle is set so that the light reflected by the multi-band dichroic mirror 140 can reach the sample through the first opening 111. The light-transmitting surface of the multi-wavelength dichroic mirror 140 is disposed facing the second opening 112, and the light excited by the sample passes through the incident surface and then continues to pass through the light-transmitting surface, and exits through the second opening 112.

With this arrangement, a set of optical elements (ie, multi-band filter 130 and multi-band dichroic mirror 140) in a light cube can be used to provide light sources of different wavelengths, so that the structure of the fluorescent lighting device is simple and beneficial Simplify the overall structure of the microscopic imaging system and reduce its cost.

In an embodiment, the second optical axis Y is perpendicular to the first optical axis X.

With this arrangement, the light path can be simpler, and the design difficulty of the fluorescent lighting device 00 and the manufacturing process difficulty are lower.

In other embodiments, the first optical axis X and the second optical axis Y can also be set to other crossing angles, which can be set according to the actual requirements of the fluorescent lighting device 00, which is not limited in the embodiment of the present application.

With reference to FIGS. 1 and 2, in an embodiment, the multi-color fluorescent light source 120 includes a multi-color LED light source 1200.

With this arrangement, using the LED light source, the multi-color fluorescent light source 120 can have a better light output effect and lower cost, thereby helping to reduce the cost of the fluorescent lighting device 00 and the microscopic imaging system.

In other embodiments, the multi-color fluorescent light source 120 may also use other types of light sources known to those skilled in the art, which is not described in detail or limited in the embodiment of the present application.

2, in an embodiment, the multi-color LED light source 1200 includes at least two different colors of monochromatic LED light sources. In FIG. 2, a first color light source 121, a second color light source 122, and a third color light source 123 are shown respectively. Out.

With this arrangement, the multi-color LED light source 1200 can provide a variety of different colors of fluorescence.

Exemplarily, the number of monochromatic LED light sources of each color can be 1, 2, or more; monochromatic LED light sources of various colors can be randomly arranged, can be arranged in an orderly sequence, can be uniform or uneven The alternate arrangement of, can be flexibly set according to the actual needs of the fluorescent lighting device 00, which is not limited in the embodiment of the present application.

3 or 4, in an embodiment, the fluorescent lighting device 00 further includes a condenser lens 150; the condenser lens 150 is disposed in the optical path between the multi-color fluorescent light source 120 and the first multi-band filter 130 .

With this configuration, the condenser lens 150 can be used to converge the light emitted from the multi-color fluorescent light source 120 to the light incident surface of the multi-wavelength filter 130, thereby improving the light utilization rate of the multi-color fluorescent light source 120.

Exemplarily, the condenser lens 150 may be a convex lens.

3 or 4, in an embodiment, the fluorescent lighting device 00 further includes an auxiliary filter structure 160; along the second optical axis Y, the auxiliary filter structure 160 is disposed on the light transmission of the multi-band dichroic mirror 140 side.

With this arrangement, the auxiliary filter structure 160 can be used to filter the light transmitted by the multi-band dichroic mirror 140, so as to obtain the fluorescence that is actually required.

Referring to FIG. 3, in an embodiment, the auxiliary filter structure 160 includes a second multi-band filter 161; the second multi-band filter 161 is disposed inside the housing 110 and covers the second opening 112.

With this arrangement, while achieving light filtering, the auxiliary filter structure 160 can be simple in structure and small in volume. This is beneficial to simplify the overall structure of the fluorescent illumination light source 00.

4, in an embodiment, the auxiliary filter structure 160 includes a filter wheel 162; the filter wheel 162 is disposed outside the housing 110, and the filter wheel (162) is rotated to enable different filters of the filter wheel 162 The effective position covers the second opening 112. The filter turntable 162 is a turntable type filter device. A plurality of filters (such as 6 or 8) are provided on the turntable according to requirements, and the position of the filter is the effective position of the filter. The filter wheel 162 can be rotated so that a certain filter of the plurality of filters provided on the wheel covers the second opening 112 and filters the light transmitted by the multi-band dichroic mirror 140.

With this arrangement, while achieving light filtering, the auxiliary filter structure 160 can be arranged independently of the housing 110, so that the internal structure of the housing 110 is less, and the entire housing can be smaller; and it is convenient to use the housing 110 and its internal structure as An independent whole body can be disassembled and assembled independently from the auxiliary filter structure 160, so that the disassembly and assembly are convenient, which is beneficial to realize the replacement of partial structural parts, and is beneficial to reduce the maintenance cost of the fluorescent lighting device 00 and the microscopic imaging system.

The fluorescent lighting device 00 provided by the embodiment of the present application includes a multi-color fluorescent light source 120, a condenser lens 150 (for example, a convex lens), a first multi-band filter 130, a second multi-band filter 161, and a multi-band dichroic mirror 140. The above-mentioned components are all integrated in a filter cube (ie, housing 110), and the housing 110 includes a first opening 111 and a second opening 112. Among them, the multi-color fluorescent light source 120, the condenser lens 150 (for example, a convex lens), the first multi-band filter 130, and the multi-band dichroic mirror 140 are on the same axis (the axis is parallel to the first optical axis X), and Arranged in sequence; the second multi-band filter 161 is located on another axis, the axis is determined by the second multi-band filter 161 and the multi-band dichroic mirror 140, and is parallel to the second optical axis Y (that is, with the first An optical axis X crosses); the first opening 111 is located on the side of the multi-wavelength dichroic mirror 140 away from the second multi-wavelength filter 161, and the second opening 112 is located on the second multi-wavelength filter 161 away from the multi-wavelength two-way One side of the color mirror 140. Among them, the multi-color fluorescent light source 120 may be a multi-color LED light source 1200. According to different types of samples to be tested, or different types of fluorescent substances in the samples, the multi-color fluorescent light source 120 may emit corresponding LED fluorescence. The condenser lens 150 (for example, a convex lens) is arranged between the multi-color fluorescent light source 120 (for example, the multi-color LED light source 1200) and the first multi-band filter 130, and the condenser lens 150 (for example, a convex lens) is arranged for the multi-color fluorescent light source. The fluorescence emitted by 120 (for example, the multi-color LED light source 1200) is focused. The first multi-band filter 130 is located between the condenser lens 150 (for example, a convex lens) and the multi-band dichroic mirror 140, and the first multi-band filter 130 is arranged to be used for the multi-color fluorescent light source 120 (for example, a multi-color LED light source). 1200) the specific fluorescence emitted by the sample is filtered, and the second multi-band filter 161 is configured to filter the emitted light excited by the sample. The multi-band dichroic mirror 140 is configured to reflect the specific fluorescence emitted by the multi-color fluorescent light source 120 (for example, the multi-color LED light source 1200) and filtered by the first multi-band filter 130, as well as those excited by the sample. Emit light.

In an embodiment, the fluorescent lighting device 00 includes a multi-color fluorescent light source 120, a condenser lens 150 (for example, a convex lens), a first multi-band filter 130, a multi-band dichroic mirror 140, and a filter wheel 162, wherein, The multi-color fluorescent light source 120, the condenser lens 150 (for example, a convex lens), the first multi-band filter 130, and the multi-band dichroic mirror 140 are all integrated in a filter cube (that is, the housing 110), and the filter wheel 162 It is disposed outside the housing 110, and the housing 110 includes a first opening 111 and a second opening 112. Wherein, the multi-color fluorescent light source 120, the condenser lens 150 (for example, a convex lens), the first multi-band filter 130, and the multi-band dichroic mirror 140 are on the same axis (the axis is parallel to the first optical axis X), and Arranged in order; the filter wheel 162 is located on another axis, the axis is determined by the filter wheel 162 and the multi-band dichroic mirror 140, and is parallel to the second optical axis Y (that is, crosses the first optical axis X); An opening 111 is located on the side of the multi-band dichroic mirror 140 away from the filter wheel 162, and the second opening 112 is located on the side of the filter wheel 162 close to the multi-band dichroic mirror 140. The condenser lens 150 (for example, a convex lens) is disposed between the multi-color fluorescent light source 120 (for example, the multi-color LED light source 1200) and the first multi-band filter 130, and the first multi-band filter 130 is located on the condenser lens 150 (for example, Between the convex lens) and the multi-band dichroic mirror 140.

In the implementation scheme of the multi-color LED light source provided by the embodiment of the present application, multiple LED light sources of different colors are arranged in the overall structure of the multi-color LED light source, which can provide a variety of different colors of fluorescence. The single color LED light source of each color can be one, two or more, and can be arranged randomly, in an orderly sequence, or unevenly and evenly alternately.

In the fluorescent lighting device 00 provided by the embodiment of the present application, the first multi-band filter 130, the second multi-band filter 161, and the filter wheel 162 are different from the traditional filter which can only filter for one type of fluorescence. It can filter a variety of fluorescences that are realized or actually required, so as to provide a variety of fluorescence with concentrated wavelength bands. The multi-band dichroic mirror 140 is different from the traditional dichroic mirror which can only reflect one kind of fluorescence, and it can reflect multiple kinds of fluorescence. In this way, while realizing that it can provide a variety of different wavelength bands of fluorescence, the overall structure of the fluorescent lighting device 00 is relatively simple and the cost is low.

An embodiment of the present application also provides a microscopic imaging system, which includes the fluorescent lighting device provided in any of the foregoing embodiments.

Exemplarily, referring to FIG. 5, the microscopic imaging system 20 further includes: a sample stage 210, an objective lens 220, and an image sensor 230; a sample stage 210, an objective lens 220, a multi-band dichroic mirror 140, and an image sensor 230 along the second The optical axis Y is set in sequence.

Among them, the specific fluorescence emitted by the multi-color LED light source (shown as the multi-color fluorescent light source 120) is focused by a condenser lens 150 (for example, a convex lens), and filtered by the first multi-band filter 130, and then passes through a multi-band dichroic The mirror 140 reflects, passes through the objective lens 220, and enters the sample on the sample stage 210; the fluorescent substance in the sample is excited by the specific fluorescence and emits emission light, and the emitted light passes through the objective lens 220, the multi-band dichroic mirror 140, and the auxiliary filter structure 160 (such as the second multi-wavelength filter 161 or the filter wheel 162, only the second multi-wavelength filter 161 is taken as an example in FIG. 5), it enters the image sensor 230 and generates a corresponding image. Thus, fluorescence microscopy imaging is completed.

Referring to FIG. 5, in an embodiment, the microscopic imaging system 20 further includes a white light source 240; the white light source 240 is disposed on the side of the sample stage 210 away from the objective lens 220.

With this arrangement, the white light source 240 can be used to perform bright field imaging to obtain a bright field image.

In other embodiments, the microscopic imaging system 20 may also include other structural components known to those skilled in the art, which are not described in detail or limited in the embodiment of the present application.

In other embodiments, the fluorescent lighting device 00 may also be applied to other lighting scenes known to those skilled in the art, which is not described in detail or limited in the embodiment of the present application.

According to the fluorescent lighting device and the microscopic imaging system provided by the embodiments of the present application, a filter cube is provided, and the filter cube includes a housing having a first opening and a second opening, and the first opening and the second opening are arranged opposite to each other; The multi-color fluorescent light source is fixed inside the housing; the first multi-wavelength filter is arranged on the light-emitting side of the multi-color fluorescent light source; the multi-wavelength dichroic mirror is arranged on the first multi-wavelength filter along the first optical axis The side away from the multi-color fluorescent light source; after being reflected by the multi-band dichroic mirror, the light travels along the second optical axis; the second optical axis passes through the first opening and the second opening, and the second optical axis is connected to the first optical axis Cross; can simplify the structure of the fluorescent lighting device and reduce its cost while providing a variety of fluorescent light sources of different wavelengths.

Claims (11)

1. A fluorescent lighting device includes a filter cube (10), and the filter cube (10) includes:

   A housing (110), the housing (110) has a first opening (111) and a second opening (112), the first opening (111) and the second opening (112) are arranged opposite to each other;

   The multi-color fluorescent light source (120) is fixed inside the housing (110);

   The first multi-band filter (130) is arranged on the light exit side of the multi-color fluorescent light source (120);

   A multi-band dichroic mirror (140) is arranged on a side of the first multi-band filter (130) away from the multi-color fluorescent light source (120) along the first optical axis (X);

   After being reflected by the multi-band dichroic mirror (140), the light propagates along the second optical axis (Y); the second optical axis (Y) passes through the first opening (111) and the second An opening (112), the second optical axis (Y) intersects the first optical axis (X).
2. The device according to claim 1, wherein the second optical axis (Y) is perpendicular to the first optical axis (X).
3. The device according to claim 1, wherein the multi-color fluorescent light source (120) comprises a multi-color LED light source (1200).
4. The device according to claim 3, wherein the multi-color LED light source (1200) comprises at least two monochromatic LED light sources of different colors.
5. The device according to claim 1, further comprising a condenser lens (150); the condenser lens (150) is disposed on the multi-color fluorescent light source (120) and the first multi-band filter (130) In the light path between.
6. The device according to claim 1, further comprising an auxiliary filter structure (160); along the second optical axis (Y), the auxiliary filter structure (160) is disposed on the multi-band dichroic mirror ( 140) of the transparent side.
7. The device according to claim 6, wherein the auxiliary filter structure (160) comprises a second multi-band filter (161);

   The second multi-band filter (161) is arranged inside the housing (110) and covers the second opening (112).
8. The device according to claim 6, wherein the auxiliary filter structure (160) comprises a filter turntable (162);

   The filter turntable (162) is arranged outside the housing (110), and the filter turntable (162) is rotated so that different effective positions of the filter turntable (162) cover the second Opening (112).
9. The device according to claim 5, wherein the condenser lens (150) is a convex lens.
10. A microscopic imaging system, comprising the fluorescent lighting device according to any one of claims 1-9; the microscopic imaging system further comprising: a sample stage (210), an objective lens (220) and an image sensor (230);

    The sample stage (210), the objective lens (220), the multi-band dichroic mirror (140) and the image sensor (230) are sequentially arranged along the second optical axis (Y).
11. The system according to claim 10, further comprising a white light source (240);

    The white light source (240) is arranged on a side of the sample stage (210) away from the objective lens (220).

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