A low-cost immunofluorescence optical detection system

Abstract

The utility model relates to the field of fluorescence detection technology, concretely relates to a low -cost immunofluorescence optical detection system, including at least one fluorescent excitation light path and a fluorescent collection light path, each fluorescent excitation light path includes the light source and the rod lens that are fixed in the oblique upper side of the sample to be measured from top to bottom in turn, and the fluorescent collection light path includes the spherical mirror, the optical filter and the fluorescence collection device that are fixed in the upper side of the sample to be measured from bottom to top in turn, the excitation light of light source generates and irradiates to the sample to be measured after focusing by the rod lens, and the fluorescence that the sample to be measured is excited generates and gathers in the fluorescence collection device after focusing by the spherical mirror and filtering by the optical filter. The low -cost immunofluorescence optical detection system of the utility model uses only the rod lens and the spherical mirror in the optical full path, and its cost is far lower than the plano -convex lens or bi -convex lens used in the conventional design, and through the one -way focusing of the rod lens, the excitation light of LED can be focused to irradiate the sample to be measured effectively, and the excitation power is guaranteed.

Description

Technical Field

[0001] This utility model relates to the field of fluorescence detection technology, specifically to a low-cost immunofluorescence optical detection system. Background Technology

[0002] The basic principle of immunofluorescence technology is to organically combine the high specificity of antigen-antibody reactions with the sensitive detectability of fluorescence. It uses fluorescent substances as tracers to label antigens or antibodies, creating specific reagents for detecting the corresponding antibodies or antigens. Specifically, a specific light source is used to excite the fluorescent substance in the antigen-antibody conjugate. The excited fluorescent substance emits fluorescence, which can be observed or quantitatively detected by an analyzer to detect and locate the function of a specific substance. Due to its advantages of accuracy, specificity, sensitivity, and speed, it is widely used in the fields of medicine, biology, and pharmacy.

[0003] With the continuous development and progress of science and technology, more immunofluorescence technologies are being applied to the field of clinical testing and analysis, and more immunofluorescence detection instruments are entering the market. Generally, the core component of an immunofluorescence detection instrument is the optical detection assembly, or simply the optical path. The optical path consists of multiple components, including a light source generator, lenses, filters, and a fluorescence acquisition device for detection, as well as structural components that connect and fix these components.

[0004] Existing commonly used optical designs share the same receiver; the difference lies in the transmitter, specifically the light-emitting device. Laser diodes can be used, characterized by high power, and their emitted light spot can be adjusted into a straight line shape via a module. The disadvantage is the limited selection of wavelengths; for example, the 365nm wavelength is currently unavailable on the market. Light-emitting diodes (LEDs) can also be used, but their power is relatively low, and their light is not focused. Therefore, multiple sets of LEDs need to be used simultaneously to meet the excitation requirements of the fluorescence detection line.

[0005] The fluorescence detection line emits fluorescence, which passes through a fluorescence entrance aperture. The aperture size is chosen primarily to block the excitation light. The fluorescence then passes through a first plano-convex lens for collimation. After collimation, the fluorescence passes through a filter for filtration, and then through a second plano-convex lens for focusing. The focused spot is concentrated on the photosensitive area of ​​the silicon photodiode, thus detecting the fluorescence intensity. Finally, the concentration of the detected substance is calculated. Conventional methods require plano-convex lenses, which are relatively expensive. Summary of the Invention

[0006] In order to overcome the shortcomings and deficiencies of the existing technology, the purpose of this utility model is to provide a low-cost immunofluorescence optical detection system.

[0007] The objective of this invention is achieved through the following technical solution: a low-cost immunofluorescence optical detection system, comprising at least one fluorescence excitation optical path and one fluorescence acquisition optical path. Each fluorescence excitation optical path includes a light source and a rod lens, which are tilted and fixed from top to bottom above the sample to be tested. The fluorescence acquisition optical path includes a spherical lens, a filter, and a fluorescence acquisition device, which are fixed from bottom to top directly above the sample to be tested. The excitation light generated by the light source is focused by the rod lens and then irradiates the sample to be tested. The fluorescence generated by the excited sample is focused by the spherical lens and filtered by the filter and then collected in the fluorescence acquisition device.

[0008] Furthermore, the light source is a light-emitting diode, and the fluorescence collection device is a silicon photodiode.

[0009] Furthermore, the excitation light is violet light with a wavelength of 365 nm, and the fluorescence has a wavelength of 610 nm.

[0010] Furthermore, the light source has a first emission aperture, and the lateral size of the light spot formed after the excitation light passes through the first emission aperture is larger than the length of the fluorescence detection line on the sample to be tested.

[0011] Furthermore, the first exit hole is a square hole with a side length of 1-1.2 mm.

[0012] Furthermore, the rod lens has a second exit hole, and the excitation light is focused unidirectionally by the rod lens. After passing through the second exit hole, the longitudinal dimension of the light spot formed is equal to the width of the fluorescence detection line on the sample to be tested.

[0013] Furthermore, the excitation light irradiates the sample to be tested, exciting the fluorescent material on the fluorescence detection line of the sample, causing the fluorescent material to emit fluorescence of the corresponding wavelength.

[0014] Furthermore, the spherical mirror has an entrance aperture and a third exit aperture. The fluorescence is focused through the entrance aperture in each vector direction of the spherical mirror and then enters the filter through the third exit aperture.

[0015] Furthermore, the filter has a fourth emission aperture. Fluorescence is filtered by the filter and then focuses on the fluorescence collection device after passing through the fourth emission aperture. The fluorescence collection device detects the intensity of the fluorescence to determine the concentration of the sample to be tested.

[0016] Furthermore, there are two fluorescence excitation light paths, which are symmetrically fixed above and to the sides of the sample to be tested, and the angle between the fluorescence excitation light path and the fluorescence acquisition light path is 30-60°.

[0017] The beneficial effects of this invention are as follows: the low-cost immunofluorescence optical detection system of this invention only uses rod lenses and spherical lenses in the entire optical path, and its cost is far lower than that of plano-convex lenses or biconvex lenses used in conventional designs. Moreover, through the unidirectional focusing of the rod lens, the excitation light of the LED can be effectively focused to illuminate the sample to be tested, and its excitation power can be guaranteed. Attached Figure Description

[0018] Figure 1 This is a perspective view of the present invention.

[0019] Figure 2 This is a perspective view of the present invention after the front shell has been removed.

[0020] Figure 3 This is a schematic diagram of the detection optical path of this utility model.

[0021] The attached figures are labeled as follows: 1. Sample to be tested; 11. Fluorescence detection line; 2. Light source; 21. First emission aperture; 3. Rod lens; 31. Second emission aperture; 4. Spherical lens; 41. Entrance aperture; 42. Third emission aperture; 5. Filter; 51. Fourth emission aperture; 6. Fluorescence collection device; 7. Housing; 71. Front housing; 72. Detailed Implementation

[0022] To facilitate understanding by those skilled in the art, the following description is provided in conjunction with embodiments and appendices. Figure 1-3 The present invention will be further described below. The content mentioned in the embodiments is not intended to limit the present invention.

[0023] See Figure 1-3 A low-cost immunofluorescence optical detection system includes at least one fluorescence excitation optical path and one fluorescence acquisition optical path. Each fluorescence excitation optical path includes a light source 2 and a rod lens 3, which are tilted and fixed from top to bottom above the sample 1. The fluorescence acquisition optical path includes a spherical lens 4, a filter 5, and a fluorescence acquisition device 6, which are fixed from bottom to top directly above the sample 1. The excitation light generated by the light source 2 is focused by the rod lens 3 and then irradiates the sample 1. The fluorescence generated by the excited sample 1 is focused by the spherical lens 4 and filtered by the filter 5 and then collected in the fluorescence acquisition device 6.

[0024] The low-cost immunofluorescence optical detection system of this invention uses only a rod lens 3 and a spherical lens 4 in the entire optical path. Its cost is far lower than that of the plano-convex lens or biconvex lens used in conventional designs. Moreover, through the unidirectional focusing of the rod lens 3, the excitation light of the LED can be effectively focused to illuminate the sample 1 to be tested, and its excitation power can be guaranteed.

[0025] Specifically, the sample to be tested 1 can be a chromatography membrane loaded with the analyte. The chromatography membrane has a fluorescence detection line 11, and the fluorescence detection line 11 contains a specifically bound fluorescent substance and the analyte.

[0026] In this embodiment, the light source 2 is a light-emitting diode (LED). Specifically, the light source 2 uses a φ3 LED, which emits light of a corresponding wavelength. The wavelength is selected based on the type of fluorescent marker; here, a 365nm violet LED is chosen. The LED emits a conical beam of light from its emitting point, with the main power concentrated within a cross-sectional angle of 17°.

[0027] In this embodiment, the light source 2 has a first emission aperture 21. After the excitation light passes through the first emission aperture 21, the lateral size of the light spot formed is larger than the length of the fluorescence detection line 11 on the sample 1. Optionally, the first emission aperture 21 is a square aperture with a side length of 1-1.2 mm. To achieve the best performance of this invention, the first emission aperture 21 is a square aperture with a side length of 1.1 mm. This aperture size is mainly used to adjust the lateral size of the light spot that is finally irradiated on the chromatography membrane, i.e., the length of the light spot. The fluorescence detection line 11 on the chromatography membrane is 1*4 mm. By adjusting the first emission aperture 21, the length of the light spot irradiated on the chromatography membrane is made slightly greater than 4 mm.

[0028] In this embodiment, the rod lens 3 has a second exit aperture 31. The excitation light is unidirectionally focused by the rod lens 3. After passing through the second exit aperture 31, the longitudinal dimension of the light spot formed is equal to the width of the fluorescence detection line 11 on the sample 1 to be tested. After passing through the first exit aperture 21, the light enters the rod lens 3. The conical light column is unidirectionally focused by the rod lens 3, focusing the light into a light-strip-like shape, i.e., a straight line, so that the light irradiates the chromatography membrane with a longitudinal dimension of about 1 mm. After being unidirectionally focused by the rod lens 3, the light then irradiates the chromatography membrane through the second exit aperture 31. The size of this aperture must be selected to prevent the excitation light from entering the ball lens 4, which would lead to detection errors.

[0029] In this embodiment, the excitation light irradiates the sample 1 to be tested, exciting the fluorescent material on the fluorescence detection line 11 of the sample 1 to emit fluorescence of a corresponding wavelength. The excitation light also irradiates the chromatography membrane, exciting the fluorescent material on the fluorescence detection line 11 to emit fluorescence of a corresponding wavelength, here 610 nm.

[0030] In this embodiment, the spherical mirror 4 has an entrance aperture 41 and a third exit aperture 42. The fluorescence passes through the entrance aperture 41, is focused in all vector directions by the spherical mirror 4, and then enters the filter 5 through the third exit aperture 42. Excitation light, due to diffuse reflection from the chromatography film, and fluorescence both pass through the entrance aperture 41. The size of the entrance aperture 41 is chosen to allow sufficient fluorescence to pass through and a sufficiently small amount of diffusely reflected excitation light to pass through, thus achieving an ideal signal-to-noise ratio. Light passing through the entrance aperture 41 enters the spherical mirror 4, which focuses light in all vector directions at 360°, similar to the principle of a plano-convex lens or biconvex lens, but with a shorter focal length and stronger focusing ability. The light focused by the spherical mirror 4 passes through the third exit aperture 42 and then enters the filter 5. The filter 5 is a narrow-pass filter, with a center wavelength of 610 nm and a half-width at half-maximum of 8 nm. Filter 5 blocks the diffusely reflected excitation light, allowing only 610nm fluorescence to pass through. The selection of filter 5 directly affects the signal-to-noise ratio. A narrow-pass filter with a half-width of 8nm meets the detection requirements of immunofluorescence chromatography.

[0031] In this embodiment, the filter 5 has a fourth emission aperture 51. Fluorescence is filtered by the filter 5 and then focused onto the fluorescence collection device 6 after passing through the fourth emission aperture 51. The fluorescence collection device 6 detects the intensity of the fluorescence to determine the concentration of the sample 1 to be tested. Specifically, the fluorescence collection device 6 is a silicon photodiode. After passing through the filter 5 and the fourth emission aperture 51, the light is directly focused onto the silicon photodiode. The intensity of the fluorescence is detected by the silicon photodiode to determine the concentration of the substance to be tested. The dimensions of the third emission aperture 42 and the fourth emission aperture 51 are not specifically specified; they are mainly used to fix the spherical mirror 4 and the filter 5.

[0032] In this embodiment, there are two fluorescence excitation light paths, which are symmetrically fixed above and to the sides of the sample 11 to be tested. The angle between the fluorescence excitation light path and the fluorescence acquisition light path is 30-60°.

[0033] In this embodiment, the optical detection system further includes a housing 7. The light source 2, bar lens 3, spherical lens 4, filter 5, and fluorescence acquisition device 6 are all installed inside the housing 7. The housing 7 includes a front shell 71 and a rear shell 72 that interlock with each other, facilitating the disassembly and replacement of internal components. A sample placement area for placing the sample 1 to be tested is provided at the bottom of the housing 7. Through holes are provided at the bottom of the housing 7 corresponding to the positions of the second exit hole 31 and the entrance hole 41. The optical detection system also includes a control and processing system, which is used to control the start and stop of the optical detection system and process the acquired fluorescence signals to obtain detection results.

[0034] The above embodiments are preferred implementations of this utility model. In addition, this utility model can also be implemented in other ways. Any obvious substitutions without departing from the concept of this utility model are within the protection scope of this utility model.

Claims

1. A low-cost immunofluorescence optical detection system, comprising at least one fluorescence excitation optical path and one fluorescence acquisition optical path, characterized in that: Each fluorescence excitation optical path includes a light source and a bar mirror that are tilted and fixed above the sample from top to bottom. The fluorescence acquisition optical path includes a spherical mirror, a filter, and a fluorescence acquisition device that are fixed directly above the sample from bottom to top. The excitation light generated by the light source is focused by the bar mirror and then irradiates the sample. The fluorescence generated by the excited sample is focused by the spherical mirror and filtered by the filter and then collected in the fluorescence acquisition device.

2. The low-cost immunofluorescence optical detection system according to claim 1, characterized in that: The light source is a light-emitting diode, and the fluorescence collection device is a silicon photodiode.

3. The low-cost immunofluorescence optical detection system according to claim 1, characterized in that: The excitation light is violet light with a wavelength of 365 nm, and the fluorescence has a wavelength of 610 nm.

4. The low-cost immunofluorescence optical detection system according to claim 3, characterized in that: The light source has a first emission aperture, and the lateral size of the light spot formed after the excitation light passes through the first emission aperture is larger than the length of the fluorescence detection line on the sample to be tested.

5. The low-cost immunofluorescence optical detection system according to claim 4, characterized in that: The first exit hole is a square hole with a side length of 1-1.2 mm.

6. The low-cost immunofluorescence optical detection system according to claim 4, characterized in that: The rod mirror has a second exit hole. The excitation light is focused unidirectionally by the rod mirror and, after passing through the second exit hole, the longitudinal dimension of the light spot is equal to the width of the fluorescence detection line on the sample to be tested.

7. The low-cost immunofluorescence optical detection system according to claim 6, characterized in that: The excitation light irradiates the sample to be tested, exciting the fluorescent material on the fluorescence detection line of the sample, causing the fluorescent material to emit fluorescence of the corresponding wavelength.

8. The low-cost immunofluorescence optical detection system according to claim 7, characterized in that: The spherical mirror has an entrance aperture and a third exit aperture. The fluorescence is focused through the entrance aperture in each vector direction of the spherical mirror and then enters the filter through the third exit aperture.

9. The low-cost immunofluorescence optical detection system according to claim 8, characterized in that: The filter has a fourth emission aperture. Fluorescence is filtered by the filter and then focuses on the fluorescence collection device after passing through the fourth emission aperture. The fluorescence collection device detects the intensity of the fluorescence to determine the concentration of the sample to be tested.

10. The low-cost immunofluorescence optical detection system according to claim 1, characterized in that: The number of fluorescence excitation optical paths is two, and the two fluorescence excitation optical paths are symmetrically fixed above and to the sides of the sample to be tested. The angle between the fluorescence excitation optical path and the fluorescence acquisition optical path is 30-60°.

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