Fluorescence imaging system

Abstract

The utility model relates to the technical field of optical imaging, in particular to a fluorescence imaging system. The imaging device comprises an imaging unit and an object carrying unit located on an imaging path of the imaging unit, a rectangular imaging view of a camera in the imaging unit on the object carrying unit serves as an imaging area, and the imaging area is divided into an imaging area A and an imaging area B along an auxiliary line perpendicular to the midpoint of the long side of the imaging area. The system further comprises two groups of LED light sources which are symmetrically distributed along the imaging path, and the two groups of LED light sources are respectively arranged on one side, close to the A imaging area, of the imaging path and one side, close to the B imaging area, of the imaging path, and two construction lines which are parallel to the imaging path and respectively pass through the central point of the A imaging area and the central point of the B imaging area are taken as two reference lines; and the midpoints of the line segments between the lens of the camera and the organism to be imaged in the two reference lines are respectively used as an irradiation point a and an irradiation point b. The utility model provides a fluorescence imaging system which not only is low in cost, but also can uniformly irradiate organisms to be imaged.

Description

Technical Field

[0001] The utility model relates to the technical field of optical imaging, and specifically relates to a fluorescence imaging system. Background Art

[0002] Fluorescence imaging is to illuminate a sample with excitation light to excite fluorescent molecules in the sample, and then use a high-sensitivity camera to capture the fluorescence signal emitted by the sample.

[0003] In the field of fluorescence imaging, the excitation light currently used is generally laser. As recorded in the text with the Chinese patent publication number CN109142305B and the title of "Two-Photon Excitation Time-Lapse Detection Fluorescence Imaging Analysis Method and Equipment for Living Animals", a femtosecond laser is used as the excitation light source.

[0004] Although the laser light has the advantage of high purity, the laser band is relatively narrow, which makes it impossible to use a single laser when different band excitation lights are required. In addition, when excited by laser, due to the limitation of the emission angle of the laser itself, the excitation field of view is also relatively narrow.

[0005] This form of laser excitation has a good excitation effect in microscopic imaging with a field of view less than about 1 centimeter. However, in macroscopic imaging where the long side length of the field of view of the box or chamber containing the sample is about 10 to 30 centimeters during the imaging process, a single laser cannot meet the requirement of uniform irradiation of the biological sample to be imaged. Obviously, multiple lasers need to be arranged during specific implementation, resulting in a significant increase in the cost of the device, so it needs to be solved urgently. Summary of the Utility Model

[0006] In order to avoid and overcome the technical problems existing in the prior art, the utility model provides a fluorescence imaging system, which not only has low cost, but also can uniformly irradiate the biological sample to be imaged.

[0007] To achieve the above object, the utility model provides the following technical solutions:

[0008] A fluorescence imaging system includes an imaging unit and a loading unit located on the imaging path of the imaging unit. Taking the rectangular imaging field of view of the camera in the imaging unit on the loading unit as the imaging area, the imaging area is divided along the auxiliary line perpendicular to the midpoint of the long side of the imaging area to form an A imaging area and a B imaging area. It also includes two groups of LED light sources symmetrically distributed along the imaging path, and the two groups of LED light sources are respectively arranged on one side of the imaging path adjacent to the A imaging area and on the other side adjacent to the B imaging area. Taking two construction lines parallel to the imaging path and passing through the center point of the A imaging area and the center point of the B respectively as two reference lines, and taking the midpoints of the line segments between the camera lens and the biological object to be imaged in the two reference lines as the a irradiation point and the b irradiation point respectively, the irradiation paths of the two groups of LED light sources are respectively inclined towards the a irradiation point and the b irradiation point and extend to the biological object to be imaged.

[0009] As a further solution of the present utility model: the installation inclination angle of the LED light source is adjustable.

[0010] As a further solution of the present utility model: a light source filter is installed at the irradiation end of the LED light source.

[0011] As a further solution of the present utility model: the position of the imaging unit on its imaging path is adjustable.

[0012] As a further solution of the present utility model: it further includes a housing, which is divided by a partition into an imaging chamber and a sample chamber arranged in sequence from top to bottom. The imaging unit and the loading unit are respectively arranged in the imaging chamber and the sample chamber; the middle part of the partition forms a light source bottom plate for installing the LED light source, and an imaging through hole is provided on the plate body of the light source bottom plate located on the imaging path.

[0013] As a further solution of the present utility model: mounting holes for positioning and installing the LED light source are provided on the plate surface of the light source bottom plate.

[0014] As a further solution of the present utility model: a first support unit for supporting the camera body is fixed inside the imaging chamber. A lens and a filter wheel are installed at the acquisition end of the body, and the filter wheel is fixed in the imaging chamber through a second support unit.

[0015] As a further solution of the present utility model: the loading unit includes a loading platform fixed at the bottom of the sample chamber.

[0016] As a further solution of the present utility model: a light-shielding cover covering the periphery of the camera is fixed inside the housing.

[0017] Compared with the prior art, the beneficial effects of the present utility model are:

[0018] 1. An LED light source is used as the emission light source of the excitation light. Compared with the form of using a laser as the light source, it has the advantages of lower cost, a wider wavelength band of the excitation light, and a larger field of view. Two groups of LED light sources are symmetrically distributed along the imaging path and are respectively inclined to point at the a illumination point and the b illumination point, so that the spot combinations of the two groups of LED light sources irradiated on the loading unit form two overlapping circles; it can effectively broaden the illumination area in the length direction of the imaging field of view, especially suitable for multi-throughput sample imaging, improving the imaging quality; in addition, it can also make the biological sample to be imaged on the loading unit receive uniform illumination, ensuring that the fluorescent molecules on the biological sample to be imaged are fully excited.

[0019] 2. The installation inclination angle of the LED light source is adjustable to adjust the irradiation path of the LED light source when the positions of the a illumination point and the b illumination point caused by the biological sample to be imaged at different heights change.

[0020] 3. The position of the imaging unit on its imaging path is adjustable to adjust the size of the imaging field of view.

[0021] 4. A light source filter is installed at the irradiation end of the LED light source; a lens and a filter wheel are installed at the acquisition end of the camera body. Through double filtering correction, it can effectively separate the autofluorescence of the organism, significantly improve the signal-to-noise ratio, and realize the detection and accurate quantification of weak target fluorescence signals. Description of the Drawings

[0022] Figure 1 It is a schematic structural diagram of the first embodiment in the imaging chamber of the present utility model.

[0023] Figure 2 It is a schematic structural diagram of the present utility model.

[0024] Figure 3 It is a schematic structural diagram of the imaging chamber of the present utility model including a light-shielding cover.

[0025] Figure 4 It is a schematic structural diagram of the LED light source of the present utility model.

[0026] Figure 5 It is a schematic structural diagram of the second embodiment in the imaging chamber of the present utility model.

[0027] Figure 6 It is a schematic structural diagram of the first embodiment of the light source bottom plate of the present utility model.

[0028] Figure 7 It is a schematic structural diagram of the second embodiment of the light source bottom plate of the present utility model.

[0029] Figure 8 It is a schematic structural diagram of the imaging area of the present utility model.

[0030] Figure 9 This is a schematic diagram of the spot distribution structure formed by the LED light source in the present utility model.

[0031] In the figure: 10, housing; 11, imaging chamber; 12, sample chamber; 13, partition; 131, light source base plate; 1311, imaging through hole; 1312, mounting hole; 14, light-shielding cover; 20, camera; 21, lens; 22, fuselage; 23, filter wheel; 30, stage; 40, LED light source; 41, light source filter; 50, first support unit; 60, second support unit. Specific embodiments

[0032] Next, the technical solutions in the embodiments of the present utility model will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present utility model. Obviously, the described embodiments are only a part of the embodiments of the present utility model, rather than all the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present utility model without creative efforts shall fall within the protection scope of the present utility model.

[0033] For ease of understanding, the specific structure and working mode of the present utility model are further described below in conjunction with the accompanying drawings:

[0034] The specific structure of the present utility model is referred to Figures 1-9 As shown, its main structure includes an imaging unit and a stage unit located on the imaging path of the imaging unit.

[0035] During the molecular fluorescence imaging process, it is necessary to excite the fluorescent molecules of the biological sample to be imaged on the stage unit through the excitation light. To ensure uniform excitation of the fluorescent molecules, the present application has studied and analyzed the irradiation points of the excitation light, and obtained the irradiation points with the best irradiation effect. As Figure 8 shown, taking the rectangular imaging field of the camera 20 in the imaging unit on the stage unit as the imaging area, this imaging area is divided into an A imaging area and a B imaging area along the auxiliary line perpendicular to the midpoint of the long side of the imaging area; taking two construction lines parallel to the imaging path and passing through the center points of the A imaging area and the B center point respectively as two reference lines, and taking the midpoints of the line segments between the camera 20 lens 21 and the biological sample to be imaged in the two reference lines as the a irradiation point and the b irradiation point respectively.

[0036] Furthermore, as Figure 1 shown, the device also includes two groups of LED light sources 40 symmetrically distributed along the imaging path, and the two groups of LED light sources 40 are respectively arranged on one side of the imaging path adjacent to the A imaging area and one side adjacent to the B imaging area. Specifically, the irradiation paths of the two groups of LED light sources 40 are respectively inclined to point to the a irradiation point and the b irradiation point and extend to the biological sample to be imaged.

[0037] This application uses the LED light source 40 as the emission light source for the excitation light. Compared with the form of using a laser as the light source, it has the advantages of lower cost, a wider wavelength band of the excitation light, and a larger field of view.

[0038] In addition, the two groups of LED light sources 40 are symmetrically distributed along the imaging path and are respectively inclined to point at the a irradiation point and the b irradiation point, as Figure 9 shown, so that the spot combinations of the two groups of LED light sources 40 irradiated on the sample-carrying unit form two overlapping circles, which can effectively broaden the irradiation area in the length direction of the imaging field of view. It is especially suitable for multi-flux sample imaging to improve the imaging quality. In addition, it can also ensure that the biological samples to be imaged on the sample-carrying unit are evenly irradiated, ensuring that the fluorescent molecules on the biological samples to be imaged are fully excited.

[0039] Based on the LED light source pointing at the b irradiation point, this application also provides Comparative Example 1 with an outward offset of the irradiation angle by 5°, and Comparative Example 2 with an outward offset of the irradiation angle by 15°. In Comparative Example 1, although both have good fluorescent molecule excitation effects, the excitation degree of the fluorescent molecules on the outer carrier is significantly higher than that on the inner carrier. In Comparative Example 2, the fluorescent molecules on the carrier are not even well excited.

[0040] It is worth mentioning that during actual implementation, as Figure 1 and Figure 5 shown, in each of the two groups of LED light sources 40, each LED light source 40 can be set to at least two lamp heads with different irradiation light wavelengths, and in the two groups of LED light sources 40, the lamp heads corresponding to the same wavelength are symmetrically distributed along the imaging path.

[0041] On the above basis, the installation inclination angle of the LED light source 40 is adjustable to adjust the irradiation path of the LED light source 40 when the positions of the a irradiation point and the b irradiation point caused by biological samples to be imaged at different heights change. Further, the position of the imaging unit on its imaging path is adjustable to adjust the size of the imaging field of view. Of course, in response to the changes in the positions of the a irradiation point and the b irradiation point caused by the change of the imaging field of view, it is also necessary to cooperate with the inclination angle adjustment of the LED light source 40 to achieve the adaptive change of its irradiation path.

[0042] On the above basis, as Figure 4 shown, a light source filter 41 is installed at the irradiation end of the LED light source 40, which can filter the light emitted by the LED light source 40. In addition, as Figure 1 shown, a lens 21 and a filter wheel 23 are installed at the acquisition end of the camera 20 body 22, which also filter the light collected by the camera 20. Through double filtering correction, the autofluorescence of the organism can be effectively separated, the signal-to-noise ratio can be significantly improved, and the detection and accurate quantification of weak target fluorescence signals can be realized.

[0043] On the basis above, as Figures 1-3 shown, in the present application, the outer shell 10 serves as the mounting carrier for the imaging unit, the sample-carrying unit, and the LED light source 40. The outer shell 10 is divided by a partition 13 to form an imaging chamber 11 and a sample chamber 12 which are arranged successively from top to bottom. The imaging unit and the sample-carrying unit are respectively arranged in the imaging chamber 11 and the sample chamber 12; the middle part of the partition 13 forms a light source bottom plate 131 for mounting the LED light source 40, and an imaging through hole 1311 is provided on the plate body of the light source bottom plate 131 located on the imaging path.

[0044] In addition, mounting holes 1312 for positioning and mounting the LED light source 40 are formed on the plate surface of the light source bottom plate 131, which facilitates the symmetric distribution installation of the two groups of LED light sources 40 during the installation process. Specifically, as Figure 6 and Figure 7 shown, the number of the mounting holes 1312 matches the number of lamp heads that the LED light source 40 needs to be provided with.

[0045] Furthermore, as shown in the figure, a first support unit 50 for supporting the body 22 of the camera 20 is fixed inside the imaging chamber 11. A lens 21 and a filter wheel 23 are installed at the acquisition end of the body 22, and the filter wheel 23 is fixed in the imaging chamber 11 through a second support unit 60, so as to stably support the body 22 and the filter wheel 23 respectively.

[0046] On the basis above, the sample-carrying unit includes a sample stage 30 fixed at the bottom of the sample chamber 12. During actual implementation, an anesthetic mechanism is further provided on the sample stage 30 to anesthetize the biological to be imaged on the sample stage 30.

[0047] In addition, a light-shielding cover 14 covering the outer periphery of the camera 20 is fixed inside the outer shell 10, which further reduces the interference of external light during the acquisition process of the camera 20.

[0048] It is worth mentioning that, in addition to being independently used as a fluorescence imaging system, the present utility model can also be combined with other imaging systems to form a multimodal optical imaging system.

[0049] Certainly, for those skilled in the art, the details of the above exemplary embodiments are not limiting to the present utility model, and it also includes the same or similar structures that can be implemented in other specific forms without departing from the spirit or basic characteristics of the present utility model. Therefore, from any point of view, the embodiments should be regarded as exemplary and non-limiting. The scope of the present utility model is defined by the appended claims rather than the above description. Therefore, all changes falling within the meaning and scope of the equivalent elements of the claims are intended to be included in the present utility model. Any reference signs in the claims should not be regarded as limiting the claimed rights.

[0050] In addition, it should be understood that although this specification is described according to embodiments, not every embodiment only contains an independent technical solution. This narrative way of the specification is only for clarity. Those skilled in the art should regard the specification as a whole, and the technical solutions in each embodiment can also be appropriately combined to form other embodiments that can be understood by those skilled in the art.

[0051] The technologies, shapes, and structures not detailedly described in the present utility model are all well-known technologies.

Claims

1. A fluorescence imaging system, comprising an imaging unit and a sample stage unit located on the imaging path of the imaging unit, characterized in that, Taking the rectangular imaging field of view of the camera (20) in the imaging unit on the loading unit as the imaging area, the imaging area is divided along the auxiliary line perpendicular to the midpoint of the long side of the imaging area to form an A imaging area and a B imaging area. It also includes two groups of LED light sources (40) symmetrically distributed along the imaging path, and the two groups of LED light sources (40) are respectively arranged on one side of the imaging path adjacent to the A imaging area and on one side adjacent to the B imaging area. Taking the two construction lines parallel to the imaging path and passing through the center point of the A imaging area and the center point of the B respectively as two reference lines, and taking the midpoints of the line segments between the lens (21) of the camera (20) and the biological object to be imaged in the two reference lines as the a irradiation point and the b irradiation point respectively, the irradiation paths of the two groups of LED light sources (40) are respectively inclined towards the a irradiation point and the b irradiation point and extend to the biological object to be imaged.

2. The fluorescence imaging system according to claim 1, characterized in that, The installation inclination angle of the LED light source (40) is adjustable.

3. A fluorescence imaging system according to claim 1 or 2, characterized in that, A light source filter (41) is installed at the irradiation end of the LED light source (40).

4. A fluorescence imaging system according to claim 1 or 2, characterized in that, The position of the imaging unit on its imaging path is adjustable.

5. A fluorescence imaging system according to claim 1 or 2, characterized in that, It also includes a housing (10), and the housing (10) is divided by a partition (13) to form an imaging chamber (11) and a sample chamber (12) arranged in sequence from top to bottom. The imaging unit and the loading unit are respectively arranged in the imaging chamber (11) and the sample chamber (12); the middle plate body of the partition (13) constitutes a light source bottom plate (131) for installing the LED light source (40), and an imaging through hole (1311) is provided on the plate body of the light source bottom plate (131) located on the imaging path.

6. The fluorescence imaging system according to claim 5, wherein, Mounting holes (1312) for positioning and installing the LED light source (40) are formed on the plate surface of the light source bottom plate (131).

7. A fluorescence imaging system according to claim 5, characterized in that, A first support unit (50) for supporting the body (22) of the camera (20) is fixed inside the imaging chamber (11). A lens (21) and a filter wheel (23) are installed at the acquisition end of the body (22), and the filter wheel (23) is fixed in the imaging chamber (11) through a second support unit (60).

8. A fluorescence imaging system according to claim 5, characterized in that, The loading unit includes a loading table (30) fixed at the bottom of the sample chamber (12).

9. A fluorescence imaging system according to claim 5, characterized in that, A light-shielding cover (14) covering the outer periphery of the camera (20) is fixed inside the housing (10).

Images