阅读说明：本技术 多色荧光激发及检测装置 (Multicolor fluorescence excitation and detection device ) 是由 马勃 游杰颖 梁骞 于 2019-08-12 设计创作，主要内容包括：本发明公开一种多色荧光激发及检测装置,包含至少一发光模块、卡匣、液态镜片模块及至少一检测模块。发光模块提供具有特定波长范围的照明光。卡匣包含检测芯片,检测芯片包含多个检测槽,环绕设置于检测芯片的外围,每一检测槽内容置对应的荧光试样,且检测槽包含第一壁面及第二壁面。照明光穿透第一壁面以照射检测槽内的荧光试样,并激发荧光试样以产生荧光信号穿透第二壁面。液态镜片模块包含至少一液态镜片贴附设置于第二壁面上,当荧光信号穿透第二壁面及液态镜片时可被增强。检测模块接收增强后的荧光信号,并将之转换为电信号。(The invention discloses a multicolor fluorescence excitation and detection device, which comprises at least one luminous module, a cassette, a liquid lens module and at least one detection module. The light emitting module provides illumination light having a specific wavelength range. The cartridge comprises a detection chip, the detection chip comprises a plurality of detection grooves which are arranged around the periphery of the detection chip, a corresponding fluorescent sample is arranged in each detection groove, and the detection grooves comprise a first wall surface and a second wall surface. The illumination light penetrates the first wall surface to irradiate the fluorescent sample in the detection groove, and excites the fluorescent sample to generate a fluorescent signal to penetrate the second wall surface. The liquid lens module comprises at least one liquid lens which is attached to the second wall surface, and the fluorescence signal can be enhanced when penetrating through the second wall surface and the liquid lens. The detection module receives the enhanced fluorescence signal and converts it into an electrical signal.)

1. A multicolor fluorescence excitation and detection device comprising:

At least one light-emitting module for providing an illuminating light with a specific wavelength range;

A cassette, which comprises a detection chip, wherein the detection chip comprises a plurality of detection grooves arranged around the periphery of the detection chip, each detection groove is used for accommodating a corresponding fluorescent sample and comprises a first wall surface and a second wall surface, and the illumination light penetrates through the first wall surface to irradiate the fluorescent sample in the detection groove and excite the fluorescent sample to generate a fluorescent signal to penetrate through the second wall surface;

A liquid lens module, including at least one liquid lens attached to the second wall surface, for enhancing the fluorescence signal when the fluorescence signal penetrates through the second wall surface and the liquid lens; and

And the at least one detection module receives the enhanced fluorescence signal and converts the fluorescence signal into an electric signal.

2. The multicolor fluorescence excitation and detection device according to claim 1, wherein the light emitting module is disposed adjacent to the first wall surface with the optical axis of the light emitting module aligned to the first wall surface, and the detection module is disposed adjacent to the second wall surface with the optical axis of the detection module aligned to the second wall surface.

3. The multicolor fluorescence excitation and detection device according to claim 1, wherein the liquid lens module further comprises a first lens attached to the first wall surface and being a condensing lens.

4. The multicolor fluorescence excitation and detection device according to claim 1, wherein the liquid lens module further comprises a second lens, a liquid layer, and a third lens, wherein the liquid layer is sandwiched between the second lens and the third lens.

5. The multicolor fluorescence excitation and detection device according to claim 4, wherein the liquid layer is filled with oil, and the surfaces of the second lens and the third lens are constituted by deformable films.

6. The multicolor fluorescence excitation and detection device according to claim 5, wherein the liquid layer filling oil has a refractive index of 1.46, and has a difference of 0.05 or more from the refractive index of the material of the cartridge.

7. The multicolor fluorescence excitation and detection device according to claim 1, wherein the first wall is a bottom wall and the second wall is a front wall.

8. The multicolor fluorescence excitation and detection device according to claim 7, wherein each of the detection cells further comprises a third wall, a fourth wall, a fifth wall and a sixth wall, wherein the third wall is opposite to the first wall, the second wall is opposite to the fourth wall, the fifth wall is opposite to the sixth wall, wherein the third wall is an upper wall, the fourth wall is a rear wall, the fifth wall is a first side wall, and the sixth wall is a second side wall.

9. The multicolor fluorescence excitation and detection device according to claim 8, wherein the third wall surface and the first wall surface are respectively composed of an optical film, wherein the thickness of the optical film of the third wall surface and the thickness of the optical film of the first wall surface are respectively between 0.1mm and 0.2mm, and the refractive index of the optical film of the third wall surface and the refractive index of the optical film of the first wall surface are respectively between 1.3 and 1.6.

10. The multicolor fluorescence excitation and detection device according to claim 1, wherein the light emitting module comprises:

A light source for generating the illumination light with a broadband wavelength; and

The first filter is arranged between the light source and the first wall surface and enables the illumination light in a first wavelength range to pass through.

11. The multicolor fluorescence excitation and detection device according to claim 10, wherein the light emitting module further comprises a first pinhole disposed between the light source and the first filter, wherein the first pinhole has an aperture size ranging from 2.0mm to 3.0 mm.

12. the multicolor fluorescence excitation and detection device according to claim 1, wherein the volume size of the detection chamber is between 10 μ L and 50 μ L.

13. The multicolor fluorescence excitation and detection device according to claim 1, wherein the detection chip is made of polycarbonate, polymethyl methacrylate or cyclic olefin copolymer, and the refractive index of the detection groove is between 1.3 and 1.6.

14. The multicolor fluorescence excitation and detection device according to claim 1, wherein the detection module comprises:

A second filter receiving the fluorescence signal and allowing the fluorescence signal in a second wavelength range to pass; and

A detector receiving the fluorescence signal having a second wavelength range and converting the fluorescence signal into the electrical signal.

15. The multicolor fluorescence excitation and detection device according to claim 14, wherein the detection module further comprises a second pinhole disposed between the second wall and the second filter, wherein the aperture size of the second pinhole is between 2.0mm and 3.0 mm.

16. The multicolor fluorescence excitation and detection device according to claim 14, wherein the detector is a photodiode, an avalanche photodiode, a photosensitive coupling element or a complementary metal oxide semiconductor.

17. The multicolor fluorescence excitation and detection device according to claim 1, wherein the multicolor fluorescence excitation and detection device is applied to at least one of the fields of qPCR, isothermal PCR, fluorescence microscopy, fluorescence spectroscopy.

Technical Field

The present invention relates to fluorescence detection devices, and particularly to a multicolor fluorescence excitation and detection device with a liquid lens module.

Background

In order to obtain a large amount of specific DNA fragments according to different needs, the scientists have been working to find an efficient way to meet their objectives, and Polymerase Chain Reaction (PCR) is one of the most economical and fast techniques to obtain billions of copies of specific DNA fragments in a short time. The PCR technique can be applied to various fields such as selective DNA isolation for genetic identification, forensic analysis for analyzing ancient DNA in archaeology, medical application of gene detection and tissue types, rapid and accurate diagnosis of infectious diseases in hospitals and research institutes, environmental hazard test for food safety, and gene fingerprint for investigating criminals, etc. The PCR technique only needs to use a small amount of DNA sample extracted from blood or tissue and add a fluorescent dye to the nucleic acid solution, so that the amplified DNA fragments can be detected by fluorescent molecules.

Dye and fluorescence detection techniques are widely used to simultaneously detect and analyze whether a target nucleic acid molecule is present in a batch of biological samples. When a fluorescent signal is emitted from a target nucleic acid molecule having a DNA binding dye or a fluorescent binding probe under excitation with light of a specific wavelength, the signal is indicative of the presence of the target nucleic acid molecule. This technique is applied to a new PCR technique called isothermal amplification method (isothermal amplification method). Among them, the optical device is an indispensable tool for detecting fluorescence emitted from a specific nucleic acid fragment in the qPCR detection technology, and the optical device must provide a light source to excite the fluorescent probe at a specific wavelength and simultaneously detect a fluorescent signal emitted from the probe.

the other alternative isothermal amplification methods rely on proteins using in vivo DNA/RNA synthesis mechanisms, rather than thermal cycling, and are dominated by enzyme activity, thus miniaturized isothermal systems have the advantages of simple design and low energy consumption.

Fluorescence detection systems have matured in a variety of fields, such as fluorescence spectroscopy and the use of fluorescence microscopy. The monochromatic light source is matched with a group of filters and optical components, and the fluorescent probe can be easily applied to monochromatic fluorescent probes. However, most of the existing fluorescence detection systems have the disadvantages of large volume, complex structure and high cost. Furthermore, most fluorescence detection systems produce a fluorescence Signal with a low Signal-to-Noise ratio (SNR). The design of the fluorescence detection device of the current portable isothermal amplification method still lags behind the biochemical technology development. Since isothermal amplification has a high tolerance for the degree of purification of a sample, most of the isothermal platforms in the market aim to create an environment with stable temperature and a detection method with medium-high throughput (throughput). More importantly, most devices using isothermal amplification are preferred in mobile detection environments, and the system needs to have a high degree of integration, so most of the current optical device designs, although widely used, are not suitable for devices using isothermal amplification.

Disclosure of Invention

An objective of the present invention is to provide a multi-color fluorescence excitation and detection device, which focuses and gathers fluorescence signals to a detection module through a liquid lens module, so as to obtain a high signal-to-noise ratio, reduce the overall size and weight, provide a portable isothermal PCR system with better performance at a lower cost, and allow the deviation of a detection tank.

It is another object of the present invention to provide a multicolor fluorescence excitation and detection device having a compact optical structure to avoid alignment and assembly difficulties, and having a high performance, robust structure to avoid losses due to light transmission and maintain a high signal-to-noise ratio.

According to one aspect of the present invention, the present invention provides a multi-color fluorescence excitation and detection device, comprising at least one light-emitting module, a cartridge, a liquid lens module and at least one detection module. Each of the light emitting modules provides illumination light having a specific wavelength range. The cassette comprises a detection chip, the detection chip comprises a plurality of detection grooves which are arranged around the periphery of the detection chip, wherein each detection groove is internally provided with a corresponding fluorescent sample, and each detection groove comprises a first wall surface and a second wall surface, wherein the illumination light penetrates through the first wall surface to irradiate the fluorescent sample in the detection groove, and excites the fluorescent sample to generate a fluorescent signal to penetrate through the second wall surface. The liquid lens module comprises at least one liquid lens which is attached to the second wall surface, so that the fluorescent signal is enhanced when penetrating through the second wall surface and the liquid lens. At least one detection module receives the enhanced fluorescence signal and converts the fluorescence signal into an electrical signal.

The invention has the beneficial effects that the invention provides the multicolor fluorescence excitation and detection device, and the multicolor fluorescence excitation and detection device can be applied to the fields of using fluorescent dyes as media, such as qPCR, isothermal PCR, fluorescence microscope, fluorescence spectrum and the like. The multicolor fluorescence excitation and detection device integrates the light-emitting module, the cassette and the detection module into a single device, so that the multicolor fluorescence excitation and detection device has the advantages of compact structure, smaller volume and lighter weight.

Drawings

FIG. 1 is a schematic structural view of a nucleic acid analysis apparatus using a multicolor fluorescence excitation and detection device according to an embodiment of the present invention.

FIG. 2 is a schematic view of the nucleic acid analyzing apparatus shown in FIG. 1, when the tank is opened.

FIG. 3 is a schematic structural diagram of a multicolor fluorescence excitation and detection device according to a preferred embodiment of the present invention.

Fig. 4 is a partially enlarged schematic view of the multicolor fluorescence excitation and detection device shown in fig. 3.

Fig. 5 is a schematic structural view of the detection tank and the liquid lens module shown in fig. 4.

Fig. 6 shows the excitation spectrum of fluorescent dye FAM.

Fig. 7 shows the emission spectrum of fluorescent dye Cy 5.

The reference numbers are as follows:

100: nucleic acid analysis apparatus

1: trough body

11: top trough body

12: bottom trough body

121: chamber

13: hinge assembly

14: containing space

2: fluid transfer unit

3: temperature control unit

4: rotary drive unit

5: liquid lens module

51: liquid lens

511: second lens

512: liquid layer

513: third lens

52: first lens

6: cartridge

61: reagent storage body

62: detection chip

625: detection tank

623: the first wall surface

621: second wall surface

622: third wall surface

624: the fourth wall surface

63: first channel

64: the second channel

65: opening of the container

7: light emitting module

71: light source

72: first filter

73: the first needle hole

8: detection module

81: second filter

82: detector

83: second pinhole

9: multicolor fluorescence excitation and detection device

Detailed Description

Some exemplary embodiments that embody features and advantages of the invention will be described in detail in the description that follows. It is to be understood that the invention is capable of modification in various ways without departing from the scope of the invention, and that the description and drawings are to be regarded as illustrative in nature and not as restrictive.

The invention provides a multicolor fluorescence excitation and detection device suitable for a nucleic acid analysis device. Furthermore, the multi-color fluorescence excitation and detection device of the present invention is suitable for a nucleic acid analysis apparatus that combines the fluid delivery unit, the temperature control unit, the rotation driving unit and the multi-color fluorescence excitation and detection device on a single device, so that the nucleic acid analysis can be performed on the single device in real time to achieve the functions of sample purification, nucleic acid extraction, nucleic acid diffusion and nucleic acid detection.

FIG. 1 is a schematic view showing a nucleic acid analyzer using a multicolor fluorescence excitation and detection device according to an embodiment of the present invention, and FIG. 2 is a schematic view showing the nucleic acid analyzer shown in FIG. 1 with a tank opened, wherein a cassette is removed from the nucleic acid analyzer. As shown in fig. 1 and 2, the nucleic acid analysis apparatus 100 includes a tank 1, a fluid transport unit 2, a temperature control unit 3, a rotation drive unit 4, and a multicolor fluorescence excitation and detection device 9. The multi-color fluorescence excitation and detection device 9 includes a cassette 6, at least one light-emitting module 7, a liquid lens module 5, and at least one detection module 8 (see fig. 3 for the light-emitting module 7 and the liquid lens module 5). The housing 1 can be opened to install the cassette 6 therein. The fluid delivery unit 2 is connected to the housing 1 and adapted to deliver reagents in the cassette 6 for sample purification and/or nucleic acid extraction. The temperature control unit 3 is disposed in the tank 1 and adapted to provide a default temperature for nucleic acid amplification. The rotation driving unit 4 is connected to the slot 1, and can rotate the cassette 6 in the slot 1 by default, in one embodiment, the rotation driving unit 4 can hold the cassette 6. At least one light emitting module 7 and at least one detection module 8 are arranged on the tank body 1, each light emitting module 7 comprises at least one optical assembly for laser, and each detection module 8 comprises at least one optical assembly for detection, and is used for nucleic acid detection or sample reaction detection.

In one embodiment, the tank 1 includes a top tank 11 and a bottom tank 12. The top tank 11 and the bottom tank 12 are connected by a hinge (hinge)13, but not limited thereto. The bottom channel 12 has a cavity 121 specifically designed to receive the cassette 6 therein. The top channel 11 may be opened so that the cassette 6 may be placed in the cavity 121 of the bottom channel 12. When the top tank body 11 is closed, a closed space is formed in the tank body 1. In one embodiment, the shape of the tank 1 may be, but not limited to, cylindrical, spherical, cubic, conical or olive shape, and the tank 1 may be made of, but not limited to, metal, ceramic, polymer, wood, glass, or other materials that provide good thermal insulation.

The bottom housing 12 is connected to the fluid delivery unit 2 by a tube or channel, and when the cassette 6 is installed in the bottom housing 12, the cassette 6 is locked and in close contact with the fluid delivery unit 2 to prevent leakage. For example, the cassette 6 may be locked to the bottom slot 12 by at least one locking element, wherein the locking element may include a clip (clip), but not limited thereto.

As shown in fig. 2, the cassette 6 includes a detection chip 62 and a reagent storage body 61, and the detection chip 62 is disposed on the top of the reagent storage body 61. The detecting chip 62 is a planar fluidic chip, and the detecting chip 62 includes a plurality of detecting grooves 625 (as shown in fig. 4), at least one first channel 63, and at least one second channel 64. The at least one first channel 63 is connected to the detection slot 625 via the at least one second channel 64. In one embodiment, a plurality of test wells 625 are disposed around the periphery of the test chip 62 and contain the sample or reagent for nucleic acid amplification and/or detection. For example, the detection channel 625 can be coated with samples or reagents for nucleic acid amplification and/or detection, such as reagents comprising different fluorescent dyes. The number of detection wells 625 is not limited and can be as many as 40 or more wells, so that the apparatus of the present invention can perform multiplexed (multiplexing) nucleic acid analysis. In some embodiments, the shape of the detection chip 62 is substantially circular, such that the detection chip 62 has a plurality of curved sides for alignment with the at least one detection module 8 to facilitate focusing.

The reagent storage body 61 includes a plurality of reagent tanks (not shown) for storing reagents used for sample purification and/or nucleic acid extraction. The reagent storage body 61 also includes a plurality of flow channels connected to the reagent reservoirs for fluid delivery. In one embodiment, the reagent storage body 61 may be, but is not limited to, a cylindrical body. The detecting chip 62 further includes at least one opening 65 disposed on the top surface of the detecting chip 62, and the opening 65 is aligned and communicated with at least one reagent groove of the reagent storage body 61 for adding the sample into the cartridge 6.

Fig. 3 is a schematic structural diagram of a multicolor fluorescence excitation and detection device according to a preferred embodiment of the invention, and fig. 4 is an enlarged structural diagram of a part of the multicolor fluorescence excitation and detection device shown in fig. 3. As shown in fig. 1, 2, 3 and 4, the multicolor fluorescence excitation and detection device 9 comprises a cassette 6, at least one light-emitting module 7, a liquid lens module 5 and at least one detection module 8. The multi-color fluorescence excitation and detection device 9 may be, but is not limited to, a device including four light emitting modules 7 and four detection modules 8, wherein each light emitting module 7 is disposed in the accommodating space 14 of the bottom tank 12 (see fig. 2) and provides illumination light with a specific wavelength range.

Referring to FIG. 4, each of the detecting slots 625 includes a first wall 623, a second wall 621, a third wall 622, a fourth wall 624, a fifth wall and a sixth wall (not shown). The first wall 623 is opposite to the third wall 622, the second wall 621 is opposite to the fourth wall 624, and the fifth wall is opposite to the sixth wall. The second wall 621, the fourth wall 624, the fifth wall and the sixth wall are connected to each other and disposed between the first wall 623 and the third wall 622. In this embodiment, the first wall 623 is a bottom wall, the second wall 621 is a front wall, the third wall 622 is an upper wall, the fourth wall 624 is a rear wall, the fifth wall is a first sidewall, and the sixth wall is a second sidewall.

Fig. 5 is a schematic structural view of the detection tank and the liquid lens module shown in fig. 4. As shown in fig. 4 and 5, in the present embodiment, the liquid lens module 5 includes a liquid lens 51 and a first lens 52. The liquid lens module 5 is used for enhancing the fluorescence signal when the fluorescence signal penetrates through the second wall 621 and the liquid lens 51. The first lens 52 is a condensing lens and is attached to a first wall surface 623 (i.e., a bottom wall surface) of the detection groove 625, so that when the illumination light generated by the light emitting module 7 penetrates through the first lens 52 attached to the first wall surface 623, the illumination light can be concentrated upwards to illuminate the fluorescent sample in the detection groove 625 and excite the fluorescent sample to generate a fluorescent signal.

The liquid lens 51 is attached to a second wall 621 (i.e., a front wall) of the detection tank 625, and includes a second lens 511, a third lens 513, and a liquid layer 512 filled between the second lens 511 and the third lens 513. In this embodiment, liquid layer 512 is filled with oil, such as: mineral oil, but not limited to, can be filled with water, glue, or other liquid materials. And, in some embodiments, the oil filling in the liquid layer 512 has a refractive index of 1.46 and has a difference of at least 0.05 or more from the refractive index of the cartridge material.

In other embodiments, the second lens 511 is used for converging light in the X direction, and the third lens 513 is used for converging light in the Y direction. And, since the second lens 511, the liquid layer 512 and the third lens 513 together constitute the liquid lens 51, and the surfaces of the second lens 511 and the third lens 513 may be deformable films, such as: thin polymers or soft elastomers. Therefore, when a voltage is applied to the liquid lens 51, the curvature of the second lens 511 and/or the third lens 513 can be changed and the focal position thereof can be adjusted due to the corresponding change in the pressure or volume of the liquid layer 512. The optical curvature of the second lens 511 and the third lens 513 with the double-tapered surface design technique maximizes optical energy transmission and enhances system performance, as well as enhancing fluorescence detection at different wavelengths through the variable focal length optics. Therefore, after the fluorescence signal generated by the fluorescence sample penetrates the second wall 621 of the detection slot 625, the liquid lens 51 concentrates, enhances and converges the fluorescence signal to the detection module 8. The detection module 8 receives the enhanced fluorescence signal transmitted by the liquid lens 51, and then converts the fluorescence signal into an electrical signal.

Referring to fig. 3 and 4 again, the light emitting module 7 is disposed adjacent to the first wall surface 623 of the detection slot 625, and the optical axis of the light emitting module 7 is aligned with the first wall surface 623 of the detection slot 625, so that the first lens 52 attached to the first wall surface 623 of the detection slot 625 can receive the illumination light with a specific wavelength range emitted by the light emitting module 7. The detecting module 8 is disposed adjacent to the second wall 621 of the detecting groove 625, and the optical axis of the detecting module 8 is aligned with the second wall 621 of the detecting groove 625, so that the detecting module 8 can receive the fluorescent signal penetrating through the second wall 621 of the detecting groove 625 and the liquid lens 51.

in one embodiment, the light emitting module 7 includes a light source 71 and a first filter 72, the light source 71 can be a light emitting diode (L ED) or a laser diode (laser diode), and the light source 71 is used for generating illumination light with a wide wavelength band, the first filter 72 is disposed between the light source 71 and the first wall 623, and allows the illumination light with a specific wavelength range generated by the light source 71, such as the illumination light with the first wavelength range, to pass through and prevents the illumination light with an unwanted wavelength generated by the light source 71 from passing through.

The light emitting module 7 further includes a first pinhole 73 disposed between the light source 71 and the first filter 72, and the first pinhole 73 of the light emitting module 7 guides the illumination light generated by the light source 71 to be located on the first filter 72 and the first wall surface 623 of the detection slot 625. In some embodiments, the first needle hole 73 has a hole diameter ranging from 2.0mm to 3.0mm, but is not limited thereto.

In some embodiments, the first channel 63 is connected to the detection slot 625 via the corresponding second channel 64, wherein the first channel 63 is configured to send the sample to the detection slot 625. The cross-sectional area of the second passage 64 is preferably smaller than the cross-sectional area of the first passage 63, so that the second passage 64 has a capillary valve for passive flow control.

In an embodiment, the third wall 622 and the first wall 623 of the detection slot 625 are both made of optical films, and the thickness of the optical film on the third wall 622 and the thickness of the optical film on the first wall 623 may be between 0.1mm and 0.2mm, but not limited thereto. The refractive index of the optical film on the third wall 622 and the refractive index of the optical film on the first wall 623 may be between 1.3 and 1.6, but not limited thereto.

in some embodiments, the volume of the detecting groove 625 of the detecting chip 62 may be between 10 μ L and 50 μ L, but not limited thereto, the detecting chip 62 may be made of Polycarbonate (PC), polymethyl methacrylate (PMMA), or Cyclic Olefin Copolymer (COC), and the refractive index of the detecting groove 625 of the detecting chip 62 may be between 1.3 and 1.6, but not limited thereto.

The detection module 8 includes a second filter 81 and a detector 82. The second filter 81 is used for receiving the fluorescence signal transmitted by the liquid lens 51 and allowing the fluorescence signal of another specific wavelength range, such as: the fluorescence signal of the second specific wavelength range passes through, and the unwanted wavelengths are prevented from passing through. The detector 82 is used for receiving the fluorescence signal with the second wavelength range passing through the second filter 81 and converting the fluorescence signal into an electric signal. In one embodiment, the detector 82 may be, but is not limited to, a Photodiode (PD), an Avalanche Photodiode (APD), a Charge Coupled Device (CCD), or a complementary metal-oxide semiconductor (CMOS).

The detection module 8 further comprises a second pinhole 83, the second pinhole 83 is disposed between the second wall 621 of the detection slot 625 and the second filter 81, and the second pinhole 83 of the detection module 8 guides the fluorescence signal generated by the fluorescence sample to be located in the detection module 8. In other embodiments, the aperture of the second pinhole 83 ranges from 2.0mm to 3.0mm, but is not limited thereto.

In some embodiments, the multi-color fluorescence excitation and detection device 9 comprises a plurality of light emitting modules 7 and a plurality of detection modules 8, such as but not limited to four light emitting modules 7 and four detection modules 8, wherein the plurality of light emitting modules 7 provide illumination light of different colors to corresponding detection slots 625 for fluorescence detection, and the plurality of detection modules 8 receive corresponding fluorescence signals, so that the plurality of detection modules 8 can simultaneously detect a plurality of samples and realize multiplex detection.

Referring to fig. 6 and 7, fig. 6 shows the excitation spectrum of fluorescent dye FAM, and fig. 7 shows the excitation spectrum of fluorescent dye Cy 5. In this embodiment, FAM and Cy5 dyes are used for demonstration, and these dyes are standard fluorescent dyes, but the system of the present invention is not limited to these fluorescent dyes. As shown in FIGS. 6 and 7, the fluorescence signal and the signal-to-noise ratio detected by the present invention are higher in intensity than those of the prior art.

Table 1 shows the results of comparing the signal-to-noise ratio of two fluorescent dyes applied to the multicolor fluorescence excitation and detection device 9 of the present invention with the prior art, wherein the concentrations of the two fluorescent dyes are both 320 nM. Table 1 clearly shows that the signal-to-noise ratio of two fluorescent dyes applied to the multicolor fluorescence excitation and detection device 9 of the present invention is quite high, even as high as 77 and 30, which indicates that the sensitivity of the multicolor fluorescence excitation and detection device 9 is quite good and that the signals from the target fluorescent dyes are highly discriminative.

TABLE 1

	FAM	Cy5
			Prior art SNR _320nM	19.9	17.7
Signal-to-noise ratio of 320nM of the present invention	77.10	30.77

In one embodiment, the light emitting modules 7 are disposed in the accommodating space 14 of the bottom slot 12, and during operation, each light emitting module 7 provides effective illumination light to one of the detecting slots 625 of the cassette 6 for detection. The detection module 8 is arranged at the outer edge of the top groove body 11 to realize optical detection, so that the sample can be detected in real time in the process of nucleic acid diffusion. Once the cartridge 6 is locked, the detection module 8 is aligned with one of the detection slots 625 in the cartridge 6 so that the results of the nucleic acid analysis can be read, and the cartridge 6 is rotated so that each detection slot 625 passes through a different light emitting module 7 and detection module 8 in sequence. In one embodiment, each of the light emitting module 7 and the detection module 8 can provide illumination light and detection of a unique color to provide different colors to perform fluorescence detection, so that the nucleic acid analysis apparatus 100 can simultaneously detect multiple targets and achieve multiplexed detection.

In actual practice, there may be some deviation when the optical axis of the light emitting module 7 or the optical axis of the detecting module 8 is aligned with the detecting groove 625. The experimental results show that the signal-to-noise ratio of the system on the target fluorescent dye still keeps good performance when the deviation angle of the fluorescent sample is within ± 2 degrees, which means that the multicolor fluorescence excitation and detection device 9 receives some deviation when the optical axis of the light emitting module 7 or the optical axis of the detection module 8 is aligned in the detection slot 625. In some embodiments, the acceptable deviation angle may be between ± 3.5 degrees.

In summary, the present invention provides a multi-color fluorescence excitation and detection device, which can be applied to many fields using fluorescent dye as medium, such as qPCR, isothermal PCR, fluorescence microscope, fluorescence spectroscopy, etc. The multicolor fluorescence excitation and detection device integrates the light-emitting module, the cassette and the detection module into a single device, so that the multicolor fluorescence excitation and detection device has the advantages of compact structure, smaller volume and lighter weight. In addition, the multicolor fluorescence excitation and detection device further comprises a liquid lens module to enhance fluorescence detection signals with different wavelengths, so that the light energy transmission is maximized, and the system performance is improved. Therefore, the multicolor fluorescence excitation and detection device does not need expensive optical components, so that the cost of the multicolor fluorescence excitation and detection device is lower. Furthermore, due to the arrangement of the plurality of light emitting modules, the plurality of detection chambers, and the plurality of detection modules, the present invention can achieve multiplexed nucleic acid analysis apparatuses and multiplexed fluorescence detection. In addition, the signal-to-noise ratio of the multicolor fluorescence excitation and detection device of the present invention is high, and some deviation can be allowed when the cassette is rotated.