Aperture, illumination light source, microscopic imaging system and method

Abstract

The embodiment of the present invention provides an aperture, an illumination light source, a microscopic imaging system and method, wherein the aperture includes an aperture plate and a plurality of light holes arranged on the aperture plate, each of the light holes is fan-shaped; the light holes include a first curved arc, a second curved arc, a first straight line segment and a second straight line segment, the first end of the first straight line segment is connected to the first end of the first curved arc, the second end of the first straight line segment is connected to the second end of the second curved arc, the second end of the second curved arc is connected to the first end of the second straight line segment, the second end of the second straight line segment is connected to the second end of the first curved arc, and the length of the first curved arc is greater than the length of the second curved arc. The embodiment of the present invention increases the illumination intensity at the edge of the concave liquid surface to solve the problem of dark edges at the edge of the concave liquid surface during final imaging in the prior art, so that the brightness of the center and edge of the concave liquid surface is closer, thereby improving the imaging quality.

Description

Technical Field

[0001] The present invention relates to microscopic imaging technology, and in particular to an aperture, an illumination light source, a microscopic imaging system and a method. Background Art

[0002] As a microscopic observation device, microscopes are increasingly used in scientific research, industry and other fields. The traditional microscope light source consists of a condenser and an aperture, which focuses the light on the specimen to be observed. When using an optical objective lens to image the well plate mainly used for biological culture, the surface tension of the liquid forms a concave liquid surface, which will result in dark edges and poor image quality. Summary of the invention

[0003] The embodiments of the present invention provide an aperture, an illumination light source, a microscopic imaging system and a method to increase the illumination intensity at the edge of a concave liquid surface, so as to solve the problem of dark edges appearing at the edge of the concave liquid surface during final imaging in the prior art, so as to make the brightness of the center and edge of the concave liquid surface in the aperture plate closer, thereby improving the imaging quality.

[0004] In a first aspect, an embodiment of the present invention provides an aperture, comprising an aperture plate and a plurality of light-through holes arranged on the aperture plate, each of the light-through holes being in a fan shape;

[0005] The light-through hole includes a first curved arc, a second curved arc, a first straight segment and a second straight segment, the first end of the first straight segment is connected to the first end of the first curved arc, the second end of the first straight segment is connected to the second end of the second curved arc, the second end of the second curved arc is connected to the first end of the second straight segment, the second end of the second straight segment is connected to the second end of the first curved arc, and the length of the first curved arc is greater than the length of the second curved arc.

[0006] Optionally, the center of the first curved arc is the light through hole center of the light through hole;

[0007] The center of the light-through hole is located between the first curved arc and the second curved arc.

[0008] Optionally, the center of the second curved arc is located between the center of the light-through hole and the first curved arc.

[0009] Optionally, the center of the first curved arc is the light through hole center of the light through hole;

[0010] The second curved arc is located between the first curved arc and the center of the light-through hole.

[0011] Optionally, the center of the first curved arc is the light through hole center of the light through hole;

[0012] The aperture plate is provided with a first light hole, a second light hole, a third light hole and a fourth light hole, the line between the light hole center of the first light hole and the light hole center of the third light hole passes through the geometric center of the aperture plate, and the line between the light hole center of the second light hole and the light hole center of the fourth light hole passes through the geometric center of the aperture plate.

[0013] Optionally, a central angle of the first curved arc is greater than 90°.

[0014] In a second aspect, an embodiment of the present invention provides a lighting source, comprising the aperture described in the first aspect, and lamp beads, a first lens, a second lens, a third lens, a fourth lens and a fifth lens arranged in sequence along the optical axis, wherein the aperture is located between the third lens and the fourth lens.

[0015] In a third aspect, an embodiment of the present invention provides a microscopic imaging system, comprising the illumination light source, imaging objective lens, aperture plate and imaging unit described in the second aspect, wherein the illumination light source provides an illumination light beam, and after the illumination light beam is projected onto the aperture plate, it is imaged onto the imaging unit by the imaging objective lens.

[0016] Optionally, the radius of the first curve arc is M, the divergence angle of the illumination light beam is θ, and the vertical distance between the illumination light source and the aperture plate is N, satisfying: 2M<tanθ×N.

[0017] Optionally, the distance between the center of the second curve arc and the center of the light-transmitting hole is D, the divergence angle of the illumination light beam is θ, and the radius of the imaging area on the aperture plate is K, satisfying: D>tanθ×K.

[0018] In a fourth aspect, an embodiment of the present invention provides a microscopic imaging method, which uses the microscopic imaging system described in the third aspect to image the sample in the well plate.

[0019] An embodiment of the present invention provides an aperture, wherein the aperture of the aperture is fan-shaped, and the aperture of the aperture comprises a first curved arc, a second curved arc, a first straight line segment, and a second straight line segment. The length of the first curved arc is greater than the length of the second curved arc, so that the side of the aperture adjacent to the first curved arc has a larger light-transmitting area, and the side of the aperture adjacent to the second curved arc has a smaller light-transmitting area. When imaging the liquid surface in an aperture plate, the side of the aperture adjacent to the first curved arc corresponds to the edge of the concave liquid surface, and the side of the aperture adjacent to the second curved arc corresponds to the center of the concave liquid surface, so as to increase the illumination intensity at the edge of the concave liquid surface, so as to solve the problem of dark edges appearing at the edge of the concave liquid surface during final imaging in the prior art, so as to make the brightness of the center and edge of the concave liquid surface in the aperture plate closer, and improve the imaging quality. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1A schematic diagram of the structure of an aperture provided by an embodiment of the present invention;

[0021] Figure 2 for Figure 1 A schematic diagram of the light hole in the diaphragm shown;

[0022] Figure 3 for Figure 1 Another schematic diagram of the light hole in the diaphragm shown;

[0023] Figure 4 A schematic diagram of a figure formed after the first light through hole, the second light through hole, the third light through hole and the fourth light through hole are spliced ​​together;

[0024] Figure 5 A schematic diagram of the structure of another aperture provided by an embodiment of the present invention;

[0025] Figure 6 for Figure 5 A schematic diagram of the light hole in the diaphragm shown;

[0026] Figure 7 A schematic diagram of the structure of another aperture provided by an embodiment of the present invention;

[0027] Figure 8 A schematic diagram of the structure of an illumination light source provided by an embodiment of the present invention;

[0028] Fig. 9 A schematic structural diagram of a microscopic imaging system provided in an embodiment of the present invention. DETAILED DESCRIPTION

[0029] The present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments. It is to be understood that the specific embodiments described herein are only used to explain the present invention, rather than to limit the present invention. It should also be noted that, for ease of description, only parts related to the present invention, rather than all structures, are shown in the accompanying drawings.

[0030] Figure 1 A schematic diagram of the structure of an aperture provided by an embodiment of the present invention, Figure 2 for Figure 1 A schematic diagram of the aperture in the diaphragm shown. Figure 3 for Figure 1 Another schematic diagram of the light hole in the diaphragm shown, refer to Figure 1 , Figure 2 and Figure 3The aperture 1 includes an aperture plate 11 and a plurality of light holes 12 arranged on the aperture plate 11, and each light hole 12 is in the shape of a sector. The aperture plate 11 is the opaque part of the aperture 1, and the light hole 12 can be an opening on the aperture plate 11, and the light hole 12 is the light-transmitting part of the aperture 1. The light hole 12 includes a first curved arc A1, a second curved arc A2, a first straight line segment L1, and a second straight line segment L2. The first end of the first straight line segment L1 is connected to the first end of the first curved arc A1, the second end of the first straight line segment L1 is connected to the second end of the second curved arc A2, the second end of the second curved arc A2 is connected to the first end of the second straight line segment L2, and the second end of the second straight line segment L2 is connected to the second end of the first curved arc A1. The first curved arc A1, the second curved arc A2, the first straight line segment L1, and the second straight line segment L2 are connected to form a closed figure, and the closed figure is in the shape of a sector. The length of the first curved arc A1 is greater than the length of the second curved arc A2.

[0031] The embodiment of the present invention provides an aperture 1, wherein a light hole 12 of the aperture 1 is fan-shaped, and the light hole 12 of the aperture 1 comprises a first curved arc A1, a second curved arc A2, a first straight line segment L1, and a second straight line segment L2. The length of the first curved arc A1 is greater than the length of the second curved arc A2, so that the side of the light hole 12 adjacent to the first curved arc A1 has a larger light-transmitting area, and the side of the light hole 12 adjacent to the second curved arc A2 has a smaller light-transmitting area. When imaging the liquid surface in the aperture plate, the side of the light hole 12 adjacent to the first curved arc A1 corresponds to the edge of the concave liquid surface, and the side of the light hole 12 adjacent to the second curved arc A2 corresponds to the center of the concave liquid surface, so as to increase the illumination intensity at the edge of the concave liquid surface, so as to solve the problem of dark edges appearing at the edge of the concave liquid surface during final imaging in the prior art, so as to make the brightness of the center and edge of the concave liquid surface in the aperture plate closer, and improve the imaging quality.

[0032] For example, reference Figure 1 , Figure 2 and Figure 3 The first curve arc A1 is a circular arc, and the second curve arc A2 is a circular arc. In other embodiments, the first curve arc A1 or the second curve arc A2 may also be a non-circular arc, for example, a parabolic arc or an elliptical arc.

[0033] Optionally, refer to Figure 1 , Figure 2 and Figure 3 , the center of the first curved arc A1 is the light hole center C1 of the light hole 12. It should be noted that the light hole center C1 is not the geometric center of the light hole 12. The light hole center C1 is located between the first curved arc A1 and the second curved arc A2. The second curved arc A2 is located on the side of the light hole center C1 away from the first curved arc A1. In the embodiment of the present invention, the first curved arc A1 and the second curved arc A2 are respectively located on both sides of the light hole center C1.

[0034] Optionally, refer to Figure 1 , Figure 2 and Figure 3 , the center C2 of the second curved arc A2 is located between the center C1 of the light hole and the first curved arc A1. The center C1 of the light hole is located between the center C2 of the second curved arc A2 and the second curved arc A2. The second curved arc A2 is retracted relative to the center C1 of the light hole, thereby blocking a part of the light in the central area, so that the image formed by the concave liquid surface will not have a central bright spot, thereby improving the imaging quality.

[0035] Optionally, refer to Figure 1 , Figure 2 and Figure 3 , the center of the first curved arc A1 is the light hole center C1 of the light hole 12. The aperture plate 11 is provided with a first light hole 121, a second light hole 122, a third light hole 123 and a fourth light hole 124. The line between the light hole center C1 of the first light hole 121 and the light hole center C1 of the third light hole 123 passes through the geometric center O of the aperture plate 11, and the line between the light hole center C1 of the second light hole 122 and the light hole center C1 of the fourth light hole 124 passes through the geometric center O of the aperture plate 11. In the embodiment of the present invention, the aperture 1 includes four light holes 12.

[0036] For example, reference Figure 1 , Figure 2 and Figure 3 , wherein the center of the first light hole 121 and the center of the third light hole 123 are symmetrical about the geometric center O of the aperture plate 11, and the center of the second light hole 122 and the center of the fourth light hole 124 are symmetrical about the geometric center O of the aperture plate 11.

[0037] For example, reference Figure 1 , Figure 2 and Figure 3 , the distance between the light hole center C1 of the first light hole 121 and the geometric center O of the aperture plate 11 is equal to the distance between the light hole center C1 of the third light hole 123 and the geometric center O of the aperture plate 11. The distance between the light hole center C1 of the second light hole 122 and the geometric center O of the aperture plate 11 is equal to the distance between the light hole center C1 of the fourth light hole 124 and the geometric center O of the aperture plate 11. The distance between the light hole center C1 of the first light hole 121 and the geometric center O of the aperture plate 11 is equal to the distance between the light hole center C1 of the second light hole 122 and the geometric center O of the aperture plate 11. The straight line where the light hole center C1 of the first light hole 121 and the geometric center O of the aperture plate 11 are located is perpendicular to the straight line where the light hole center C1 of the second light hole 122 and the geometric center O of the aperture plate 11 are located.

[0038] Figure 4 Schematic diagram of a figure formed by splicing the first light hole, the second light hole, the third light hole and the fourth light hole together, refer to Figure 1-Figure 4 When the aperture provided by the embodiment of the present invention is used to photograph and image the sample on a single hole of the aperture plate, when each light hole (including the first light hole 121, the second light hole 122, the third light hole 123 and the fourth light hole 124) rotates around the geometric center O of the aperture plate 11 to the same shooting position, the same sample is photographed and imaged. When the first light hole 121, the second light hole 122, the third light hole 123 and the fourth light hole 124 are at the same shooting position, the outer edge of the figure formed by splicing the first light hole 121, the second light hole 122, the third light hole 123 and the fourth light hole 124 together is a circle. In the embodiment of the present invention, after splicing the first light hole 121, the second light hole 122, the third light hole 123 and the fourth light hole 124, a complete circular area is formed.

[0039] For example, there are 200 μl of liquid in a 96-well plate (the total height of the liquid in the 96 wells is 6 mm, and the height of the concave meniscus is 1.1 mm). At this time, the following method can be used: Figure 1 The aperture 1 shown in the figure makes the brightness of the center and edge of the concave liquid surface in the aperture plate closer, thereby improving the imaging quality. In addition, the center C1 of the light-through hole is located between the first curved arc A1 and the second curved arc A2, and the outer edge of the figure formed by splicing multiple light-through holes 12 together is circular.

[0040] Optionally, refer to Figure 1-Figure 4 , the center angle of the first curve arc A1 is greater than 90°. Figure 3 As shown, line segment B1C1 is perpendicular to line segment B2C1, the center angle of arc B1B2 is 90°, the length of first arc A1 is greater than arc B1B2, and the center angle of first arc A1 is greater than 90°, thereby ensuring that the outer edge of the figure formed by splicing the first light-through hole 121, the second light-through hole 122, the third light-through hole 123 and the fourth light-through hole 124 together is circular, and providing a splicing margin for the light-through hole 12, taking into account design and manufacturing errors, and reducing the design and manufacturing difficulty of the light-through hole 12.

[0041] Continue to refer Figure 1-Figure 3 , the distance D between the center C2 of the second curved arc A2 and the center C1 of the light-through hole satisfies: 0.8 mm ≤ D ≤ 1.2 mm. In the embodiment of the present invention, the distance D between the center C2 of the second curved arc A2 and the center C1 of the light-through hole is greater than or equal to 0.8 mm and less than or equal to 1.2 mm, so that the brightness of the center and edge of the concave liquid surface in the aperture plate are closer, thereby improving the imaging quality.

[0042] Continue to refer Figure 1-Figure 3, the radius R of the second curve arc A2 satisfies: 2.25mm≤R≤3.25mm. In the embodiment of the present invention, the radius R of the second curve arc A2 is greater than or equal to 2.25mm and less than or equal to 3.25mm, so as to improve the brightness of the edge of the concave liquid surface in the orifice plate, make the brightness of the center and edge of the concave liquid surface in the orifice plate closer, and improve the imaging quality.

[0043] Figure 5 A schematic diagram of the structure of another aperture provided by an embodiment of the present invention, Figure 6 for Figure 5 A schematic diagram of the aperture in the diaphragm shown, see Figure 5 and Figure 6 The center of the first curved arc A1 is the light hole center C1 of the light hole 12, and the second curved arc A2 is located between the first curved arc A1 and the light hole center C1. The light hole center C1 is located on the side of the second curved arc A2 away from the first curved arc A1.

[0044] For example, reference Figure 5 and Figure 6 The center C2 of the second arc A2 is located between the second arc A2 and the center C1 of the light hole, the center C1 of the light hole and the center C2 of the second arc A2 are both located on the same side of the first arc A1, and the center C1 of the light hole and the center C2 of the second arc A2 are both located on the same side of the second arc A2.

[0045] Optionally, refer to Figure 5 and Figure 6 The distance D between the center C2 of the second curved arc A2 and the center C1 of the light-through hole satisfies: 0.8 mm ≤ D ≤ 1.2 mm. The radius R of the second curved arc A2 satisfies: 2.25 mm ≤ R ≤ 3.25 mm.

[0046] For example, there are 150 μl of liquid in a 96-well plate (the total height of the liquid in the 96 wells is 5.5 mm, and the height of the concave meniscus is 1.3 mm). At this time, the following method can be used: Figure 5 The aperture 1 shown in the figure makes the center of the concave liquid surface in the well plate closer to the brightness of the edge, thereby improving the imaging quality. In addition, the second curve arc A2 is located between the first curve arc A1 and the center C1 of the light-through hole, and the outer edge of the figure formed by splicing together a plurality of light-through holes 12 is a circular ring. It should be noted that when the volume of liquid in the well plate (for example, a 96-well plate) is small (for example, less than or equal to 150 μl), the following method is used: Figure 5In the aperture 1 shown in the figure, the exact center of the concave liquid surface corresponds to the opaque area, so that the image formed by the concave liquid surface will not have a central bright spot, thereby improving the imaging quality. It should be further explained that the center of the concave liquid surface is a larger area, and the exact center of the concave liquid surface is located at the center of the area where the center of the concave liquid surface is located. The embodiment of the present invention avoids the appearance of a central bright spot on the basis of ensuring that the brightness of the center and edge of the concave liquid surface in the orifice plate is closer, thereby improving the imaging quality.

[0047] Figure 7 A schematic diagram of another aperture structure provided by an embodiment of the present invention, referring to Figure 7 The aperture plate is provided with a first light hole 121, a second light hole 122, a third light hole 123 and a fourth light hole 124. The line between the light hole center C1 of the first light hole 121 and the light hole center C1 of the third light hole 123 passes through the geometric center O of the aperture plate 11, and the line between the light hole center C1 of the second light hole 122 and the light hole center C1 of the fourth light hole 124 passes through the geometric center O of the aperture plate 11. The first light hole 121 and the third light hole 123 satisfy that the light hole center C1 is located between the first curve arc A1 and the second curve arc A2. The second light hole 122 and the fourth light hole 124 satisfy that the second curve arc A2 is located between the first curve arc A1 and the light hole center C1.

[0048] It should be noted that, in other implementations, the aperture 1 may also include other numbers of light holes 12 , and the embodiment of the present invention does not limit the number of light holes 12 . For example, the aperture 1 may also include 2, 3, 8, or 9 light holes 12 .

[0049] For example, reference Figure 1 and Figure 5 In any one of the drawings, the aperture 1 further includes a light opening 13 . Different from the fan-shaped light opening 12 , the light opening 13 is circular, and the light opening 13 is also a light-transmitting portion in the aperture 1 .

[0050] Figure 8 A schematic diagram of a lighting source according to an embodiment of the present invention is provided. Figure 8, the illumination light source 41 includes the aperture 1 in the above-mentioned embodiment, and the lamp bead 2, the first lens 31, the second lens 32, the third lens 33, the fourth lens 34 and the fifth lens 35 arranged in sequence along the optical axis, and the aperture 1 is located between the third lens 33 and the fourth lens 34. Since the illumination light source 41 provided by the embodiment of the present invention includes the aperture 1 in the above-mentioned embodiment, the illumination intensity at the edge of the concave liquid surface can be increased to solve the problem of dark edges appearing at the edge of the concave liquid surface during final imaging in the prior art, so that the brightness of the center and edge of the concave liquid surface in the orifice plate are closer, thereby improving the imaging quality. Furthermore, the first lens 31, the second lens 32 and the third lens 33 converge the light emitted by the lamp bead 2, so the aperture 1 is set between the third lens 33 and the fourth lens 34, which is conducive to selective light transmission of the converged light beam through the aperture 1, thereby improving the imaging quality.

[0051] Fig. 9 A schematic diagram of a microscopic imaging system provided by an embodiment of the present invention, referring to Fig. 9 The microscopic imaging system includes the illumination light source 41, the imaging objective lens 43, the aperture plate 42 and the imaging unit 45 in the above-mentioned embodiment. The illumination light source 41 provides an illumination beam. After the illumination beam is projected onto the aperture plate 42, it is imaged on the imaging unit 45 by the imaging objective lens 43. Since the illumination light source 41 provided in the embodiment of the present invention includes the illumination light source 41 in the above-mentioned embodiment, the illumination intensity at the edge of the concave liquid surface can be increased to solve the problem of dark edges at the edge of the concave liquid surface during final imaging in the prior art, so that the brightness of the center and edge of the concave liquid surface in the aperture plate are closer, thereby improving the imaging quality.

[0052] For example, reference Fig. 9 The aperture plate 42 is located between the illumination light source 41 and the imaging objective lens 43, and the imaging objective lens 43 is located between the aperture plate 42 and the imaging unit 45. The microscopic imaging system may further include a tube lens 44, and the tube lens 44 is located between the imaging objective lens 43 and the imaging unit 45. After the illumination light beam is projected onto the aperture plate 42, it passes through the imaging objective lens 43 and the tube lens 44 in sequence, and is imaged on the imaging unit 45.

[0053] Optionally, the radius of the first curved arc A1 is M, the divergence angle of the illumination beam is θ, and the divergence angle of the illumination beam refers to the derivative of the radius of the illumination beam to the far-field axial position. The vertical distance between the illumination light source 41 and the aperture plate 42 is N, satisfying: 2M<tanθ×N.

[0054] Optionally, the distance between the center of the second curved arc A2 and the center of the light hole C1 is D, the divergence angle of the illumination light beam is θ, and the radius of the imaging area on the aperture plate 42 is K, satisfying: D>tanθ×K. The imaging area on the aperture plate 42 refers to the imaging area of ​​the sample to be tested on the aperture plate 42 when performing microscopic imaging. For example, the aperture plate 42 includes a 96-well plate, and the 96-well plate includes 96 wells. A liquid to be tested is arranged in at least one well, and the liquid to be tested is the sample to be tested.

[0055] Exemplarily, the divergence angle of the illumination light beam is less than or equal to 15°, and the radius of the imaging area on the aperture plate 42 is less than or equal to 3 mm.

[0056] Based on the same technical concept, an embodiment of the present invention further provides a microscopic imaging method, which uses the microscopic imaging system in the above embodiment to image the sample in the well plate. The microscopic imaging method provided by the embodiment of the present invention can be performed based on the microscopic imaging system in the above embodiment, so that when the microscopic imaging method provided by the embodiment of the present invention is used, the brightness of the center and edge of the concave liquid surface in the well plate is closer, thereby improving the imaging quality.

[0057] Note that the above are only preferred embodiments of the present invention and the technical principles used. Those skilled in the art will understand that the present invention is not limited to the specific embodiments described herein, and that various obvious changes, readjustments, combinations and substitutions can be made by those skilled in the art without departing from the scope of protection of the present invention. Therefore, although the present invention has been described in more detail through the above embodiments, the present invention is not limited to the above embodiments, and may include more other equivalent embodiments without departing from the concept of the present invention, and the scope of the present invention is determined by the scope of the appended claims.

Claims

1. A diaphragm, characterized in that: It comprises an aperture plate and a plurality of light-through holes arranged on the aperture plate, each of the light-through holes is in a fan shape; The light-through hole comprises a first curved arc, a second curved arc, a first straight segment and a second straight segment, wherein a first end of the first straight segment is connected to a first end of the first curved arc, a second end of the first straight segment is connected to a first end of the second curved arc, a second end of the second curved arc is connected to a first end of the second straight segment, a second end of the second straight segment is connected to a second end of the first curved arc, and a length of the first curved arc is greater than a length of the second curved arc; The center of the first curved arc is the light hole center of the light hole; The aperture plate is provided with a first light through hole, a second light through hole, a third light through hole and a fourth light through hole, a line connecting the light through hole center of the first light through hole and the light through hole center of the third light through hole passes through the geometric center of the aperture plate, and a line connecting the light through hole center of the second light through hole and the light through hole center of the fourth light through hole passes through the geometric center of the aperture plate; When photographing and imaging the concave liquid surface in the aperture plate, the side of the light-through hole adjacent to the first curved arc corresponds to the edge of the concave liquid surface, and the side of the light-through hole adjacent to the second curved arc corresponds to the center of the concave liquid surface, thereby increasing the illumination intensity at the edge of the concave liquid surface.

2. The diaphragm according to claim 1, characterized in that The center of the first curved arc is the light hole center of the light hole; The center of the light-through hole is located between the first curved arc and the second curved arc.

3. The aperture according to claim 2, characterized in that The center of the second curved arc is located between the center of the light-through hole and the first curved arc.

4. The diaphragm according to claim 1, characterized in that The center of the first curved arc is the light hole center of the light hole; The second curved arc is located between the first curved arc and the center of the light-through hole.

5. The diaphragm according to claim 1, characterized in that The central angle of the first curved arc is greater than 90°.

6. A lighting source, characterized in that: The invention comprises the aperture as claimed in any one of claims 1 to 5, and a lamp bead, a first lens, a second lens, a third lens, a fourth lens and a fifth lens arranged in sequence along the optical axis, wherein the aperture is located between the third lens and the fourth lens.

7. A microscopic imaging system, characterized in that: It comprises the illumination light source, imaging objective lens, aperture plate and imaging unit as described in claim 6, wherein the illumination light source provides an illumination light beam, and after the illumination light beam is projected onto the aperture plate, it is imaged onto the imaging unit by the imaging objective lens.

8. The microscopic imaging system according to claim 7, characterized in that: The radius of the first curved arc is M, and the divergence angle of the illumination beam is , the vertical distance between the illumination light source and the orifice plate is N, satisfying: .

9. The microscopic imaging system according to claim 7, characterized in that: The distance between the center of the second curved arc and the center of the light hole is D, and the divergence angle of the illumination beam is , the radius of the imaging area on the orifice plate is ,satisfy: .

10. A microscopic imaging method, characterized in that: The sample in the well plate is imaged using the microscopic imaging system as described in any one of claims 7 to 9.

Images