Integrated prism-type total internal reflection fluorescence imaging system

Abstract

The utility model discloses an integration prism type total internal reflection fluorescence imaging system, including mounting support, the upside of mounting support is provided with elevating unit, the front side of elevating unit is provided with the connecting frame, the downside of connecting frame is installed with the prism through prism installation component, the mounting support is installed with laser emission head, the downside of connecting frame still is installed with Z axis displacement stage, the Z axis displacement stage is installed with the mirror subassembly corresponding with laser emission head to reflect the laser that laser emission head emitted to the prism, and the mirror subassembly is integrated to the prism through setting connecting frame and can be lifted and descended integrally along with elevating unit, and the optical path need not be reset when replacing sample, and sample replacement is more convenient.

Description

Technical Field

[0001] This utility model relates to the field of fluorescence microscopy imaging technology, specifically an integrated prism-type total internal reflection fluorescence imaging system. Background Technology

[0002] Total internal reflection illumination (TIRF) is primarily used in fluorescence microscopy imaging. Its main principle is to illuminate the sample using evanescent waves generated during total internal reflection. Because the amplitude of evanescent waves decays exponentially with increasing depth perpendicular to the interface, it can excite only fluorescent groups within a thin layer on the sample surface, suppressing noise from off-focal planes and improving image contrast.

[0003] Currently, TIRF microscopes are mainly divided into two types: objective-type TIRF and prism-type TIRF. Objective-type TIRF achieves total internal reflection by focusing the illumination beam onto the back focal plane of a high numerical aperture objective lens and changing the focusing position on the back focal plane to deflect the illumination beam. However, this method has high requirements for the numerical aperture of the objective lens and suffers from problems such as reduced contrast due to internal scattering and a small illumination field of view. Prism-type TIRF, on the other hand, has a large illumination field of view, clean evanescent waves, and a better signal-to-noise ratio because its illumination beam does not pass directly through the objective lens, thus reducing light scattering within the objective lens. It also has fewer limitations on the objective lens. However, due to its inherent characteristics, prism-type TIRF requires an additional prism on the sample to achieve total internal reflection. This means that every time the sample is changed, the prism and the corresponding mirror need to be moved, and the optical path needs to be readjusted. This is the main difficulty for the commercial application of this technology. Utility Model Content

[0004] This invention provides an integrated prism-type total internal reflection fluorescence imaging system, which can solve the problem that it is inconvenient to readjust the prism and optical path after changing the sample.

[0005] To achieve the above objectives, this utility model provides the following technical solution: an integrated prism-type total internal reflection fluorescence imaging system, including a mounting bracket. A lifting unit is provided on the upper side of the mounting bracket, and a connecting frame is provided on the front side of the lifting unit. A prism is mounted on the lower side of the connecting frame via a prism mounting assembly. A laser emitting head is mounted on the mounting bracket, and a Z-axis displacement stage is also mounted on the lower side of the connecting frame. A reflector assembly corresponding to the laser emitting head is mounted on the Z-axis displacement stage to reflect the laser emitted by the laser emitting head to the prism. By integrating the prism and reflector assembly together through the connecting frame, the system can be raised and lowered as a whole with the lifting unit. When changing samples, the optical path does not need to be reset, making sample replacement more convenient.

[0006] Preferably, the lower end of the lifting unit is connected to the mounting bracket via an XY displacement stage. The XY displacement stage allows adjustment of the displacement of the lifting unit and the connecting frame in the X and Y axes to accommodate different samples.

[0007] Preferably, the lifting unit is an electric Z-axis displacement stage, which includes a lifting slider that moves up and down on its front side. A guide post is vertically mounted on the front side of the lifting slider, and a connecting slider is slidably mounted on the guide post. The connecting slider is connected to a connecting frame, and a spring is sleeved on the guide post on the upper side of the connecting slider. The spring applies a downward force to the connecting slider. By setting the spring, the connecting slider can always be located at the lowest end, so that the contact force can always be maintained when the prism and the glass slide are in contact.

[0008] Preferably, the prism mounting assembly includes a prism mounting block, the lower end of which is provided with a prism limiting slot. The prism is embedded in the prism limiting slot and is limited by a clamping block on one side of the prism mounting block. The prism can be quickly installed and replaced by removing and installing the clamping block.

[0009] Preferably, the upper end of the prism mounting block is connected to the force sensor via a first connecting block, and the upper end of the force sensor is connected to the connecting frame via a second connecting block. This allows for the detection of the pressure when the bottom surface of the prism is in contact with the glass slide. The pressure should not exceed a set value, thereby protecting the prism and the sample from stress damage.

[0010] Preferably, a side mounting plate is vertically installed on one side of the lower end of the connecting frame, and the Z-axis displacement stage is installed on the inner side of the side mounting plate. The side mounting plate can achieve a firm installation of the Z-axis displacement stage and facilitates the connection between the Z-axis displacement stage and the reflector assembly.

[0011] Preferably, a forward-extending reflective connecting plate is installed on one side of the Z-axis displacement stage, and the reflector assembly is connected to the reflective connecting plate and located in front of the prism. The reflective connecting plate can limit the installation angle of the reflector assembly.

[0012] Preferably, the reflector assembly includes a reflector base and a reflector detachably mounted on the reflector base facing the prism side, allowing for the replacement of different reflectors as needed.

[0013] Compared with the prior art, the beneficial effects of this utility model are: By integrating the prism and reflector assemblies together using a connecting frame, they can be raised and lowered as a whole with the lifting unit. When changing samples, the optical path does not need to be reset, making sample replacement more convenient. By setting a Z-axis displacement stage, the height of the reflector assembly can be adjusted independently, thereby adjusting the angle of the laser incident into the prism, ensuring the generation of evanescent waves and the adjustment of penetration depth. By installing a spring on the lifting unit to apply downward pushing force to the connecting frame, the prism can always be kept in contact with the glass slide. Attached Figure Description

[0014] Figure 1 This is a first-view perspective three-dimensional structural diagram of the present invention; Figure 2 This is a second-view perspective three-dimensional structural diagram of the present invention; Figure 3 This is a partial first-view perspective three-dimensional structural diagram of the present invention; Figure 4 This is a partial second-view perspective three-dimensional structural diagram of the present invention; Figure 5 This is a side sectional view of the present invention; Figure 6 This is a three-dimensional structural diagram of the utility model in use.

[0015] Figure label: 1. Mounting bracket; 11. Reflector base; 2. XY displacement stage; 3. Lifting unit; 31. Connecting slider; 32. Spring; 33. Guide column; 34. Lifting slider; 4. Connecting frame; 5. Z-axis displacement stage; 51. Reflective connecting plate; 52. Mounting plate; 6. Laser emitter; 7. Reflector assembly; 8. Prism; 9. Prism mounting assembly; 91. Prism mounting block; 92. First connecting block; 93. Force sensor; 94. Second connecting block; 10. Reflector. Detailed Implementation

[0016] The technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings of the embodiments of the present invention.

[0017] This invention aims to solve the problem of inconvenience caused by the need for readjustment of the prism and optical path after sample replacement. For example... Figure 1-6As shown, the following technical solution is provided: an integrated prism-type total internal reflection fluorescence imaging system, including a mounting bracket 1. A lifting unit 3 is provided on the upper side of the mounting bracket 1, and a connecting frame 4 is provided on the front side of the lifting unit 3. A prism 8 is mounted on the lower side of the connecting frame 4 via a prism mounting assembly 9. A laser emitter 6 is mounted on the mounting bracket 1, and a Z-axis displacement stage 5 is also mounted on the lower side of the connecting frame 4. A reflector assembly 7 corresponding to the laser emitter 6 is mounted on the Z-axis displacement stage 5 to reflect the laser emitted by the laser emitter 6 to the prism 8. By setting the connecting frame 4 to integrate the prism 8 and the reflector assembly 7 together, they can be raised and lowered as a whole with the lifting unit 3. When changing samples, the optical path does not need to be reset, making sample replacement more convenient.

[0018] Specifically, such as Figure 6 As shown, the integrated prism-type total internal reflection fluorescence imaging system in this embodiment can be mounted on an inverted microscope. The laser emitter 6 is connected to the laser generator via an optical fiber. A collimating lens can be installed at the lower end of the laser emitter 6. The laser light is collimated by the collimating lens, then reflected by the reflector assembly 7 into the prism 8, achieving total internal reflection at the carrier plate and sample surface, thus forming an evanescent wave illuminating the sample. The collimating lens here can be an adjustable collimating lens, and the size of the illumination area can be adjusted by changing the position of the collimating lens and the optical fiber.

[0019] The lifting unit 3 can be an electric Z-axis displacement stage, whose core structure is a combination of a ball screw pair and a servo motor (such as the Panasonic MINAS A6 series), with a positioning accuracy of ±1μm.

[0020] The Z-axis displacement stage 5 is a mature component in existing technology and can be purchased on the market. It generally includes two displacement plates that can move relative to each other along the Z-axis. The position of the two plates is adjusted by a high-precision nut. The reflector assembly 7 is installed on one of the displacement plates, and the height of the reflector assembly 7 can be adjusted by rotating the high-precision nut.

[0021] In this embodiment, the lower end of the lifting unit 3 is connected to the mounting bracket 1 via the XY displacement stage 2. The XY displacement stage 2 allows adjustment of the displacement of the lifting unit 3 and the connecting frame 4 in the X and Y axes to accommodate different samples. The XY displacement stage 2 uses a commercially available, mature product, employing a cross slide structure and including X-axis and Y-axis guide rails. The guide rails are high-precision linear ball bearing guides (such as the THKSR12 model), with a positioning accuracy of ±5μm. Two driving methods are available: Manual mode: Fine adjustment is achieved through a high-precision nut and a precision lead screw (1mm lead). Each rotation of the high-precision nut corresponds to a 1mm displacement, and the minimum scale division is 0.01mm. Electric mode: An integrated two-phase stepper motor (such as 42BYGH40-1704A) is used, sending pulse signals through a PLC controller (not shown). The minimum step angle is 1.8°, corresponding to a displacement of 0.5μm / pulse. When it is necessary to adapt to samples of different sizes, the operator can input the sample coordinates through the touch screen (not shown), and the system will automatically control the movement of the XY displacement stage 2 to achieve precise positioning of the laser spot on the sample surface.

[0022] In this embodiment, the lifting unit 3 is an electric Z-axis displacement stage. The electric Z-axis displacement stage includes a lifting slider 34 that moves up and down on its front side. A guide post 33 is vertically installed on the front side of the lifting slider 34. A connecting slider 31 is slidably disposed on the guide post 33. The connecting slider 31 is connected to the connecting frame 4. A spring 32 is sleeved on the guide post 33 on the upper side of the connecting slider 31. The spring 32 applies a downward force to the connecting slider 31. By setting the spring 32, the connecting slider 31 can always be located at the lowest end, so that the contact force can always be maintained when the prism 8 contacts the glass slide. Specifically, two guide posts 33 can be arranged side by side, and the guide posts 33 pass vertically through the connecting slider 31.

[0023] In this embodiment, the prism mounting assembly 9 includes a prism mounting block 91. The lower end of the prism mounting block 91 is provided with a prism limiting slot. The prism 8 is embedded in the prism limiting slot and is limited by a clamping block 95 on one side of the prism mounting block 91. The prism 8 can be quickly installed and replaced by removing and installing the clamping block 95. The upper end of the prism mounting block 91 is connected to a force sensor 93 via a first connecting block 92. The upper end of the force sensor 93 is connected to a connecting frame 4 via a second connecting block 94. This allows the sensor to detect the pressure when the bottom surface of the prism 8 is in contact with the glass slide. The pressure should not exceed a set value, thereby protecting the prism and the sample from stress damage. Specifically, a spring 32 can work in conjunction with the force sensor 93. The elastic coefficient of the spring 32 is designed to be 5-10 N / mm. When the prism 8 contacts the glass slide, the force sensor 93 monitors the pressure value in real time. Taking a standard glass slide (thickness 1.0mm±0.1mm) as an example, the system controls the clamping force to be maintained within the range of 0.5-1.0N by the electric Z-axis displacement stage 3; when the pressure is too low (<0.5N), the spring 32 pushes the connecting slider 31 to move downward to increase the fit; when the pressure is too high (>1.0N), the force sensor 93 triggers the electric Z-axis displacement stage to reverse the micro-motion to prevent the glass slide from breaking or the prism 8 from undergoing stress deformation (the degree of deformation can be controlled within ≤5μm).

[0024] In this embodiment, a side mounting plate 52 is vertically installed on one side of the lower end of the connecting frame 4. The Z-axis displacement stage 5 is installed inside the side mounting plate 52. The side mounting plate 52 enables the Z-axis displacement stage 5 to be firmly installed and facilitates the connection between the Z-axis displacement stage 5 and the reflector assembly 7. A forward-extending reflective connecting plate 51 is installed on one side of the Z-axis displacement stage 5. The reflector assembly 7 is connected to the reflective connecting plate 51 and located in front of the prism 8. The reflective connecting plate 51 can limit the installation angle of the reflector assembly 7.

[0025] In this embodiment, the reflector assembly 7 includes a reflector base 11 and a reflector 10 detachably mounted on the reflector base 11 on the side facing the prism 8, and different reflectors 10 can be replaced as needed.

[0026] It should be noted that all directional indicators (such as up, down, left, right, front, back, etc.) in this utility model embodiment are only used to explain the relative positional relationship and movement of each component in a certain specific posture (as shown in the figure). If the specific posture changes, the directional indicator will also change accordingly.

[0027] Furthermore, in this utility model, descriptions involving terms such as "primary," "secondary," etc., are for descriptive purposes only and should not be construed as indicating or implying their relative importance or implicitly specifying the number of technical features indicated. Therefore, a feature defined as "primary" or "secondary" may explicitly or implicitly include at least one of those features. In the description of this utility model, "multiple" means at least two, such as two, three, etc., unless otherwise explicitly and specifically defined.

[0028] In this utility model, unless otherwise explicitly specified and limited, the terms "connection," "fixing," etc., should be interpreted broadly. For example, "fixing" can mean a fixed connection, a detachable connection, or an integral part; it can mean a mechanical connection or an electrical connection; it can mean a direct connection or an indirect connection through an intermediate medium; it can mean the internal communication of two components or the interaction between two components, unless otherwise explicitly limited. Those skilled in the art can understand the specific meaning of the above terms in this utility model according to the specific circumstances.

[0029] Furthermore, the technical solutions of the various embodiments of this utility model can be combined with each other, but only if they are based on the ability of those skilled in the art to implement them. When the combination of technical solutions is contradictory or cannot be implemented, it should be considered that such combination of technical solutions does not exist and is not within the scope of protection claimed by this utility model.

Claims

1. An integrated prism-type total internal reflection fluorescence imaging system, comprising a mounting bracket (1), characterized in that, The mounting bracket (1) is provided with a lifting unit (3) on its upper side, and a connecting frame (4) is provided on the front side of the lifting unit (3). A prism (8) is installed on the lower side of the connecting frame (4) via a prism mounting assembly (9). A laser emitting head (6) is installed on the mounting bracket (1). A Z-axis displacement stage (5) is also installed on the lower side of the connecting frame (4). A reflector assembly (7) corresponding to the laser emitting head (6) is installed on the Z-axis displacement stage (5) to reflect the laser emitted by the laser emitting head (6) to the prism (8).

2. The integrated prism-type total internal reflection fluorescence imaging system according to claim 1, characterized in that: The lower end of the lifting unit (3) is connected to the mounting bracket (1) via the XY displacement stage (2).

3. The integrated prism-type total internal reflection fluorescence imaging system according to claim 2, characterized in that: The lifting unit (3) is an electric Z-axis displacement stage. The electric Z-axis displacement stage includes a lifting slider (34) that is lifted and lowered on its front side. A guide column (33) is vertically installed on the front side of the lifting slider (34). A connecting slider (31) is slidably arranged on the guide column (33). The connecting slider (31) is connected to the connecting frame (4). A spring (32) is sleeved on the guide column (33) on the upper side of the connecting slider (31). The spring (32) applies a downward force to the connecting slider (31).

4. The integrated prism-type total internal reflection fluorescence imaging system according to claim 3, characterized in that: The prism mounting assembly (9) includes a prism mounting block (91), and a prism limiting slot is provided at the lower end of the prism mounting block (91). The prism (8) is embedded in the prism limiting slot and is limited by a clamping block (95) on one side of the prism mounting block (91).

5. The integrated prism-type total internal reflection fluorescence imaging system according to claim 4, characterized in that: The upper end of the prism mounting block (91) is connected to the force sensor (93) through the first connecting block (92), and the upper end of the force sensor (93) is connected to the connecting frame (4) through the second connecting block (94).

6. The integrated prism-type total internal reflection fluorescence imaging system according to claim 1, characterized in that: A side mounting plate (52) is vertically installed on one side of the lower end of the connecting frame (4), and the Z-axis displacement stage (5) is installed on the inner side of the side mounting plate (52).

7. The integrated prism-type total internal reflection fluorescence imaging system according to claim 6, characterized in that: A forward-extending reflective connecting plate (51) is installed on one side of the Z-axis displacement stage (5), and the reflector assembly (7) is connected to the reflective connecting plate (51) and located in front of the prism (8).

8. The integrated prism-type total internal reflection fluorescence imaging system according to claim 1, characterized in that: The mirror assembly (7) includes a mirror base (11) and a mirror (10) detachably mounted on the side of the mirror base (11) facing the prism (8).

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