Multi-channel fluorescent dye optical filter combination for FISH (fluorescence in situ hybridization) detection

Abstract

The utility model discloses a multichannel fluorescent dye optical filter combination for FISH (fluorescence in situ hybridization) detection, which comprises an excitation optical filter, an emission optical filter and a color separation optical filter, and the excitation optical filter, the emission optical filter and the color separation optical filter respectively comprise a glass substrate arranged at the bottommost layer; the glass substrate comprises a plurality of high-refractive-index coating layers and a plurality of low-refractive-index coating layers, and each high-refractive-index coating layer and each low-refractive-index coating layer are sequentially arranged on the glass substrate according to alternate sputtering; the multi-channel fluorescent dye filter combination is suitable for simultaneously detecting three fluorescent markers of enhanced cyan fluorescent protein (ECFP), fluorescein isothiocyanate (FITC) and cyan pigment 3 (CY3).

Description

Technical Field

[0001] This utility model relates to the technical field of filter structure design, and more specifically, to a multi-channel fluorescent dye filter combination for fluorescence in situ hybridization (FISH) detection. Background Technology

[0002] Fluorescence in situ hybridization (FISH) is a molecular biology technique used to detect the location of specific DNA sequences in cells or tissues. It utilizes fluorescently labeled DNA probes that bind complementary to target DNA sequences, and then observes the location and intensity of the fluorescence signal using a fluorescence microscope. Commonly used fluorescent dyes in FISH include ECFP (Enhanced Cyan Fluorescent Protein), FITC (Fluorescein Isothiocyanate), and the orange fluorescent dye CY3. These dyes play a crucial role in gene localization, chromosomal abnormality detection, and molecular interaction studies due to their unique spectral properties and biocompatibility. Each dye has different excitation and emission spectra, therefore, appropriate filters must be selected for observation. The choice of filter depends on the excitation and emission spectra of the fluorescent dye. A typical filter system includes an excitation filter, an emission filter, and a dichroic mirror. The excitation filter allows light of a specific wavelength to pass through to excite the fluorescent dye, the emission filter allows the fluorescence emitted by the dye to pass through while blocking other light, and the dichroic mirror is used to reflect the excitation light and transmit the emitted light.

[0003] In the field of fluorescence detection, traditional single-channel fluorescence filters, due to their optical design having only a single passband, are limited to detecting only a single type of fluorescent substance. When faced with the need for multi-target fluorescent substance detection, the conventional solution is to integrate multiple single-channel filters through a mechanical switching device, achieving time-sharing detection of different fluorescent substances via mechanical switching. However, this strategy suffers from low detection efficiency and high system complexity.

[0004] Therefore, there is an urgent need in the field for a multi-channel fluorescent dye filter combination for FISH detection, which should have a simple structure and efficiently achieve time-sequential detection of different fluorescent substances. Utility Model Content

[0005] This invention provides a multi-channel fluorescent dye filter combination for FISH detection, which integrates multiple independent passbands on a single filter element, enabling it to simultaneously transmit incident light of different wavelengths and generate specific emission light through the analyte.

[0006] To achieve the above objectives, this utility model provides a multi-channel fluorescent dye filter assembly for FISH detection, characterized in that it comprises:

[0007] An excitation filter is placed between the light source and the substance to be detected to filter incident light of a preset wavelength to excite the substance to be detected to generate a fluorescence signal.

[0008] A dichroic filter, tilted at a 45° angle in the path of the emitted light from the fluorescence signal generated by the substance to be detected, is used to reflect the excitation light in the fluorescence signal and transmit the emitted light; and

[0009] An emission filter is disposed between the dichroic filter and a detector for secondary filtering of emitted light in the fluorescence signal;

[0010] The structures of the excitation filter, the emission filter, and the dichroic filter all include:

[0011] A glass substrate, located at the bottom layer; and

[0012] The film structure includes multiple high-refractive-index coating layers and multiple low-refractive-index coating layers, wherein each high-refractive-index coating layer and each low-refractive-index coating layer are sequentially and alternately sputtered onto the glass substrate;

[0013] The excitation filter and the emission filter both have a film structure of 142 layers, while the color separation filter has a film structure of 80 layers.

[0014] In one embodiment of this utility model, the film structure of the excitation filter, from the glass substrate upwards, is as follows: 23.54nm H, 40.19nm L, 116.22nm H, 355.57nm L, 110.94nm H, 57.33nm L, 50.15nm H, 298.71nm L, 125.23nm H, 470.38nm L, 66.17nm H, 111.1nm L, 56.66nm H, 77.23nm L, 55.17nm H, 23.83nm L, 183.1nm H, 420.31nm L, 93.27nm H, 78.92nm L, 49.79nm L... H at 84.63nm, L at 67.55nm, L at 84.91nm, H at 164.16nm, L at 211.48nm, H at 167.1nm, L at 146.55nm, H at 45.25nm, L at 83.65nm, H at 65.12nm, L at 109.32nm, H at 64.15nm, L at 75.06nm, H at 43.17nm, L at 218.34nm, H at 171.11nm, L at 90.37nm, L at 69.4nm, L at 109.81nm, H at 68.9nm, L at 91.63nm, H at 52.96nm, L at 89.87nm, H at 60.08nm 239.92nm L, 28.05nm H, 53.8nm L, 85.51nm H, 86.75nm L, 67.41nm H, 85.34nm L, 87.55nm H, 84.89nm L, 53.22nm H, 82.75nm L, 55.25nm H, 226.15nm L, 161.65nm H, 75.91nm L, 64.75nm H, 77.83nm L, 82.07nm H, 118.06nm L, 51.15nm H, 83.26nm L, 50.78nm H, 254.57nm L, 153.59nm H, 81.67 L at 49.91nm, H at 36.76nm, H at 74.86nm, L at 127.69nm, H at 70.58nm, L at 86.22nm, H at 68.78nm, L at 68.01nm, H at 52.07nm, L at 129.7nm, H at 132.09nm, L at 53.92nm, H at 58.73nm, L at 71.07nm, H at 81.93nm, L at 55.04nm, H at 95.99nm, L at 75.04nm, H at 57.45nm, L at 97.3nm, H at 67.24nm, L at 93.98nm, H at 125.37nm, L at 78.59nm, 56.76nm H, 86.44nm L, 69.97nm H, 78.89nm L, 90.52nm H, 80.43nm L, 51.71nm H, 278.21nm L, 62.74nm H, 59.57nm L, 35.74nm H, 50.12nm L, 59.97nm H, 85.74nm L, 72.49nm H, 77.49nm L, 77.16nm H, 97.03nm L, 74.51nm H, 254.21nm L, 50.36nm H, 76.4nm L, 139.52nm H, 72.01nm L, 70.51nm H, 79.64nm The wavelengths are: L (81.15nm), H (64.34nm), H (86.01nm), L (267.11nm), H (50.44nm), L (72.28nm), H (142.76nm), L (66.76nm), H (92.33nm), L (34.93nm), L (82.34nm), H (34.01nm), H (209.16nm), L (71.32nm), H (157.25nm), L (255.69nm), H (252.61nm), L (321.93nm), H (60.01nm), L (168.03nm), H (113.59nm), and L (105.43nm), where H represents high-refractive-index materials and L represents low-refractive-index materials.

[0015] In one embodiment of this utility model, the film structure of the emission filter, from the glass substrate upwards, is as follows: H at 108.41nm, L at 398.56nm, H at 22.26nm, L at 255.34nm, H at 11.61nm, L at 70.19nm, H at 57.36nm, L at 69.51nm, H at 17.64nm, L at 59.64nm, H at 168.92nm, L at 216.85nm, H at 185.23nm, L at 137.59nm, H at 32.72nm, L at 63.19nm, H at 46.9nm, L at 212.37nm, H at 153.74nm, L at 289.56nm, and so on. 3.8nm H, 108.44nm L, 64.91nm H, 103.2nm L, 22.49nm H, 79.4nm L, 105.08nm H, 161.82nm L, 45.02nm H, 244.86nm L, 150.66nm H, 132.89nm L, 52.91nm H, 85.02nm L, 54.72nm H, 105.86nm L, 23.08nm H, 45.59nm L, 158.66nm H, 266.55nm L, 137.02nm H, 131.61nm L, 66.09nm H, 72.64nm L, 47 0.04nm H, 98.98nm L, 56.18nm H, 25.23nm L, 149.92nm H, 272.53nm L, 125.22nm H, 148.06nm L, 55.02nm H, 90.84nm L, 48.47nm H, 101.24nm L, 26.63nm H, 118.98nm L, 118.78nm H, 268.06nm L, 113.74nm H, 170.23nm L, 51.04nm H, 79.74nm L, 59.02nm H, 86.79nm L, 53.31nm H, 75.44nm L, 12 1.14nm H, 313.65nm L, 101.83nm H, 143.17nm L, 69.3nm H, 63.27nm L, 59.67nm H, 83.9nm L, 57.83nm H, 78.32nm L, 128.09nm H, 313.49nm L, 92.82nm H, 137.42nm L, 73.12nm H, 73.84nm L, 51.33nm H, 79.17nm L, 50.88nm H, 100.42nm L, 130.13nm H, 313.34nm L, 77.84nm H, 159.16nm L, 60.0.1nm H, 92.87nm L, 45.03nm H, 72.14nm L, 51.53nm H, 118.25nm L, 131.91nm H, 311.73nm L, 53.61nm H, 169.66nm L, 78.72nm H, 61.79nm L, 58.2nm H, 71.92nm L, 69.46nm H, 41.8nm L, 171.76nm H, 279.76nm L, 69.79nm H, 173.58nm L, 83.75nm H, 37.2nm L, 40.71nm H, 87.69nm L, 60.31nm H, 103.19nm L, 142 The following are the wavelengths of materials with refractive indices: 0.79nm (H), 299.22nm (L), 59.62nm (H), 205.44nm (L), 52.39nm (H), 39.99nm (L), 69.86nm (H), 28.6nm (L), 59.82nm (H), 126.8nm (L), 58.83nm (H), 434.3nm (L), 146.52nm (H), 124.48nm (L), 12.89nm (H), 41.2nm (L), 73.91nm (H), 48.92nm (L), 191.08nm (H), 426.38nm (L), 11.43nm (H), 214.49nm (L), 8.82nm (H), and 289.74nm (L). Here, H represents a high refractive index material, and L represents a low refractive index material.

[0016] In one embodiment of this utility model, the film structure of the color filter, from the glass substrate upwards, is as follows: 96.96nm H, 183.69nm L, 124.05nm H, 416.87nm L, 9.46nm H, 31.9nm L, 105.55nm H, 108.11nm L, 16.17nm H, 218.38nm L, 91.76nm H, 78.03nm L, 143.47nm H, 394.33nm L, 20.06nm H, 89.62nm L, 223.41nm H, 171.92nm... L of m, H of 154.96nm, L of 472.49nm, H of 15.38nm, L of 246nm, H of 66.72nm, L of 23.62nm, H of 130.21nm, L of 238.04nm, H of 143nm, L of 321.59nm, H of 373.71nm, L of 49.66nm, H of 155.65nm, L of 560.02nm, H of 57.31nm, L of 207.42nm, H of 26.2nm, L of 123.22nm, H of 157.38nm, L of 74.79nm, H of 134.43nm, 43 2.44nm L, 103.88nm H, 98.16nm L, 56.59nm H, 99.81nm L, 72nm H, 15.76nm L, 76.3nm H, 223.73nm L, 107.4nm H, 381.78nm L, 94.93nm H, 83.95nm L, 58.92nm H, 104.89nm L, 133.67nm H, 162.16nm L, 86.12nm H, 434.38nm L, 120.4nm H, 172.89nm L, 18.01nm H The wavelengths are: L = 126.93 nm, H = 188.18 nm, L = 119.89 nm, H = 136.13 nm, L = 334.75 nm, H = 130.18 nm, L = 34.33 nm, H = 91.53 nm, L = 320.85 nm, H = 158.68 nm, L = 529.16 nm, H = 27.96 nm, L = 64.82 nm, H = 41.25 nm, L = 54.6 nm, H = 42.05 nm, L = 236.62 nm, H = 28.77 nm, and L = 152.69 nm. Here, H represents high refractive index material and L represents low refractive index material.

[0017] In one embodiment of this utility model, the center wavelength of the excitation filter is 435±2nm, 487±2nm, and 565±2nm, and the bandwidths are 20±2nm, 15±2nm, and 20±2nm, respectively. The excitation filter has an average transmittance of >85% for light with wavelengths in the range of 425-445nm, 480-494nm, and 555-574nm, a cutoff range of 452-475nm@OD>6, and a background depth of 500-550nm@OD>6.

[0018] In one embodiment of this utility model, the center wavelength of the emission filter is 462±2nm, 530±2nm, and 605±2nm, and the bandwidths are 15±2nm, 20±2nm, and 40±2nm, respectively; the emission filter has an average transmittance of >85% for light with wavelengths in the range of 455-470nm, 520-540nm, and 586-624nm, a cutoff range of 475-515nm@OD>6, and a background depth of 547-577nm@OD>6.

[0019] In one embodiment of this utility model, the center wavelengths of the dichroic filters are 464±2nm, 530±2nm, and 606±2nm, and their bandwidths are 20±3nm, 20±2nm, and 40±2nm, respectively; the average transmittance of the dichroic filters for light with wavelengths in the range of 455-472nm, 520-542nm, and 586-626nm is >85%.

[0020] In one embodiment of this utility model, the dichroic filter has a reflectivity of >95% for excitation light and a transmittance of >92% for emitted light at an incident angle of 45°.

[0021] In one embodiment of this utility model, the multi-channel fluorescent dye filter assembly for FISH detection is characterized in that the glass substrate is made of borosilicate optical glass, the high refractive index material is tantalum pentoxide, and the low refractive index material is silicon dioxide.

[0022] This invention provides a multi-channel fluorescent dye filter combination for FISH detection. The multi-channel excitation filter, multi-channel emission filter, and multi-channel dichroic filter achieve selective transmission or reflection of multiple wavelengths through the interference effect of multiple thin film layers, significantly improving the efficiency of multicolor fluorescence imaging or multivariate Raman analysis. This not only overcomes the spectral limitations of single-channel filters but also enables real-time synchronous detection of multiple fluorescent substances, significantly improving detection efficiency and reducing overall cost. Attached Figure Description

[0023] To more clearly illustrate the technical solutions in the embodiments of this utility model or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of this utility model. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0024] Figure 1 This is a schematic diagram of the optical path of a multi-channel fluorescent dye filter combination according to an embodiment of the present invention;

[0025] Figure 2 This is a simulation design spectral curve of the excitation filter according to an embodiment of the present invention;

[0026] Figure 3 This is a spectral curve of the actual coating of the excitation filter according to an embodiment of the present invention;

[0027] Figure 4 This is a simulation design spectral curve of an emission filter according to an embodiment of the present invention.

[0028] Figure 5 This is a spectral curve of the actual coating of an emission filter according to an embodiment of the present invention.

[0029] Figure 6 This is a simulation design spectral curve of a color separation filter according to an embodiment of the present invention;

[0030] Figure 7 This is a spectral curve of the actual coating of a color-separated filter according to an embodiment of the present invention.

[0031] Explanation of reference numerals in attached figures: Figures 2-7 The horizontal axis in the figure represents wavelength (nm), and the vertical axis in the figure represents transmittance (%). Detailed Implementation

[0032] The technical solutions of the present utility model will be clearly and completely described below with reference to the accompanying drawings of the embodiments. Obviously, the described embodiments are only some embodiments of the present utility model, and not all embodiments. Based on the embodiments of the present utility model, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the protection scope of the present utility model.

[0033] Figure 1 This is a schematic diagram of the optical path of a multi-channel fluorescent dye filter combination according to an embodiment of the present invention. Figure 2 The following is a simulation design spectral curve of the excitation filter according to an embodiment of this utility model. Figure 3 This is a spectral curve of the actual coating of an excitation filter according to an embodiment of the present invention. Figure 4 The following is a simulation design spectral curve of the emission filter according to an embodiment of this utility model. Figure 5 This is a spectral curve of the actual coating of an emission filter according to an embodiment of the present invention. Figure 6 The following is a simulation design spectral curve diagram of a color separation filter according to an embodiment of this utility model. Figure 7 This is a spectral curve of the actual coating of a color-separated filter according to an embodiment of the present invention, as shown below. Figure 1 , Figure 2 , Figure 3 , Figure 4 , Figure 5 , Figure 6 and Figure 7 As shown, this utility model provides a multi-channel fluorescent dye filter assembly for FISH detection, comprising: an excitation filter, an emission filter, and a dichroic filter. The structures of the excitation filter, the emission filter, and the dichroic filter all include: a glass substrate disposed at the bottom layer; and

[0034] The glass substrate has multiple high-refractive-index coating layers and multiple low-refractive-index coating layers, wherein each high-refractive-index coating layer and each low-refractive-index coating layer are sequentially sputtered on the glass substrate.

[0035] The excitation filter is disposed between the light source and the substance to be detected, and is used to filter incident light of a preset wavelength to excite the substance to be detected to generate a fluorescence signal.

[0036] The dichroic filter is tilted at a 45° angle in the path of the emitted light from the fluorescence signal generated by the substance to be detected, so that the excitation light in the fluorescence signal is reflected and the emitted light is transmitted; and

[0037] The emission filter is disposed between the dichroic filter and a detector for secondary filtering of emitted light in the fluorescence signal;

[0038] The film structure of the excitation filter is shown in the table below, consisting of 142 layers. In the marking codes, H represents a high refractive index material and L represents a low refractive index material.

[0039]

[0040]

[0041]

[0042]

[0043]

[0044] The specific film structure of the emission filter is shown in the table below, consisting of 142 layers. In the marking codes, H represents a high refractive index material and L represents a low refractive index material.

[0045]

[0046]

[0047]

[0048]

[0049]

[0050] The specific film structure of the dichroic filter is shown in the table below. It consists of 80 layers, with H representing high refractive index material and L representing low refractive index material in the marking codes.

[0051]

[0052]

[0053]

[0054] When the broadband spectrum emitted by the composite light source is incident on the excitation filter, the excitation filter selectively transmits light of a specific excitation wavelength and effectively blocks light of other wavelengths. The transmitted excitation light acts on the analyte to generate a fluorescence signal, which includes excitation and emission light. After passing through a 45° dichroic filter, the excitation light is reflected, and the emission light is transmitted. Finally, the emission filter performs secondary purification of the fluorescence signal to ensure that the detector only receives the specific fluorescence signal with a high signal-to-noise ratio, thereby enabling the analysis of the components of the analyte. The excitation filter, the emission filter, and the dichroic filter are integrated to form a three-element system.

[0055] In this embodiment, the excitation filter, the emission filter, and the color separation filter are all produced by ion-assisted magnetron sputtering to precisely control the properties of each film layer in order to achieve a complex multi-channel structure, but this is not a limitation.

[0056] In this embodiment, the center wavelengths of the excitation filter are 435±2nm, 487±2nm, and 565±2nm, and the bandwidths are 20±2nm, 15±2nm, and 20±2nm, respectively. The excitation filter has an average transmittance of >85% for light with wavelengths in the range of 425-445nm, 480-494nm, and 555-574nm, a cutoff range of 452-475nm@OD>6, and a background depth of 500-550nm@OD>6.

[0057] In this embodiment, the transmission band of the excitation filter covers three ranges: 430-440nm (ECFP), 490-500nm (FITC), and 545-555nm (CY3).

[0058] In this embodiment, the center wavelengths of the emission filter are 462±2nm, 530±2nm, and 605±2nm, with bandwidths of 15±2nm, 20±2nm, and 40±2nm, respectively. The emission filter has an average transmittance of >85% for light with wavelengths in the ranges of 455-470nm, 520-540nm, and 586-624nm, a cutoff range of 475-515nm@OD>6, and a background depth of 547-577nm@OD>6.

[0059] In this embodiment, the emission filter is provided with three independent channels: 470-480nm (ECFP), 520-530nm (FITC), and 565-575nm (CY3).

[0060] In this embodiment, the center wavelengths of the dichroic filters are 464±2nm, 530±2nm, and 606±2nm, and their bandwidths are 20±3nm, 20±2nm, and 40±2nm, respectively; the average transmittance of the dichroic filters for light with wavelengths in the range of 455-472nm, 520-542nm, and 586-626nm is >85%.

[0061] In this embodiment, at an incident angle of 45°, the dichroic filter has a reflectivity of >95% for excitation light and a transmittance of >92% for emitted light.

[0062] In this embodiment, the glass substrate material is borosilicate optical glass, the high refractive index coating layer material is tantalum pentoxide, and the low refractive index coating layer material is silicon dioxide.

[0063] In this embodiment, the multi-channel fluorescent dye filter combination for FISH detection can simultaneously detect ECFP cyan dye, FITC green dye, and CY3 orange fluorescent dye.

[0064] Those skilled in the art will understand that the accompanying drawings are merely schematic diagrams of one embodiment, and the modules or processes shown in the drawings are not necessarily essential for implementing this utility model.

[0065] Those skilled in the art will understand that the modules in the apparatus of the embodiments can be distributed in the apparatus of the embodiments as described in the embodiments, or they can be located in one or more devices different from this embodiment with corresponding changes. The modules of the above embodiments can be combined into one module, or they can be further divided into multiple sub-modules.

[0066] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of this utility model, and not to limit it; although the utility model has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of this utility model.

Claims

1. A multi-channel fluorescent dye filter assembly for FISH detection, characterized in that, include: An excitation filter is placed between the light source and the substance to be detected to filter incident light of a preset wavelength to excite the substance to be detected to generate a fluorescence signal. A dichroic filter, tilted at a 45° angle in the path of the emitted light from the fluorescence signal generated by the substance to be detected, is used to reflect the excitation light in the fluorescence signal and transmit the emitted light; and An emission filter is disposed between the dichroic filter and a detector for secondary filtering of emitted light in the fluorescence signal; The structures of the excitation filter, the emission filter, and the dichroic filter all include: A glass substrate, located at the bottom layer; and The film structure includes multiple high-refractive-index coating layers and multiple low-refractive-index coating layers, wherein each high-refractive-index coating layer and each low-refractive-index coating layer are sequentially and alternately sputtered onto the glass substrate; The excitation filter and the emission filter both have a film structure of 142 layers, while the color separation filter has a film structure of 80 layers.

2. The multi-channel fluorescent dye filter assembly for FISH detection according to claim 1, characterized in that, The film structure of the excitation filter, from the glass substrate upwards, is as follows: H at 23.54 nm, L at 40.19 nm, H at 116.22 nm, L at 355.57 nm, H at 110.94 nm, L at 57.33 nm, H at 50.15 nm, L at 298.71 nm, H at 125.23 nm, L at 470.38 nm, H at 66.17 nm, L at 111.1 nm, H at 56.66 nm, L at 77.23 nm, H at 55.17 nm, L at 23.83 nm, H at 183.1 nm, L at 420.31 nm, H at 93.27 nm, L at 78.92 nm, H at 49.79 nm, and H at 84.63 nm. L, 67.55nm H, 84.91nm L, 164.16nm H, 211.48nm L, 167.1nm H, 146.55nm L, 45.25nm H, 83.65nm L, 65.12nm H, 109.32nm L, 64.15nm H, 75.06nm L, 43.17nm H, 218.34nm L, 171.11nm H, 90.37nm L, 69.4nm H, 109.81nm L, 68.9nm H, 91.63nm L, 52.96nm H, 89.87nm L, 60.08nm H, 239.92nm L, 2 8.05nm H, 53.8nm L, 85.51nm H, 86.75nm L, 67.41nm H, 85.34nm L, 87.55nm H, 84.89nm L, 53.22nm H, 82.75nm L, 55.25nm H, 226.15nm L, 161.65nm H, 75.91nm L, 64.75nm H, 77.83nm L, 82.07nm H, 118.06nm L, 51.15nm H, 83.26nm L, 50.78nm H, 254.57nm L, 153.59nm H, 81.67nm L, 49.91nm H at m, L at 36.76nm, H at 74.86nm, L at 127.69nm, H at 70.58nm, L at 86.22nm, H at 68.78nm, L at 68.01nm, H at 52.07nm, L at 129.7nm, H at 132.09nm, L at 53.92nm, H at 58.73nm, L at 71.07nm, H at 81.93nm, L at 55.04nm, H at 95.99nm, L at 75.04nm, H at 57.45nm, L at 97.3nm, H at 67.24nm, L at 93.98nm, H at 125.37nm, L at 78.59nm, H at 56.76nm, 86.44nm L, 69.97nm H, 78.89nm L, 90.52nm H, 80.43nm L, 51.71nm H, 278.21nm L, 62.74nm H, 59.57nm L, 35.74nm H, 50.12nm L, 59.97nm H, 85.74nm L, 72.49nm H, 77.49nm L, 77.16nm H, 97.03nm L, 74.51nm H, 254.21nm L, 50.36nm H, 76.4nm L, 139.52nm H, 72.01nm L, 70.51nm H, 79.64nm L, 81 The following are the wavelengths of materials with refractive indices: 0.15nm (H), 64.34nm (L), 86.01nm (H), 267.11nm (L), 50.44nm (H), 72.28nm (L), 142.76nm (H), 66.76nm (L), 92.33nm (H), 34.93nm (L), 82.34nm (H), 34.01nm (L), 209.16nm (H), 71.32nm (L), 157.25nm (H), 255.69nm (L), 252.61nm (H), 321.93nm (L), 60.01nm (H), 168.03nm (L), 113.59nm (H), and 105.43nm (L). Here, H represents a high refractive index material, and L represents a low refractive index material.

3. The multi-channel fluorescent dye filter assembly for FISH detection according to claim 1, characterized in that, The film structure of the emission filter, from the glass substrate upwards, is as follows: H at 108.41 nm, L at 398.56 nm, H at 22.26 nm, L at 255.34 nm, H at 11.61 nm, and H at 70.19 nm. L, 57.36nm H, 69.51nm L, 17.64nm H, 59.64nm L, 168.92nm H, L at 216.85nm, H at 185.23nm, L at 137.59nm, H at 32.72nm, 63.19nm L, 46.9nm H, 212.37nm L, 153.74nm H, 289.56nm L, 143.8nm H, L at 108.44nm, H at 64.91nm, L at 103.2nm, H at 22.49nm, 79.4nm L, H at 105.08nm, L at 161.82nm, H at 45.02nm, L at 244.86nm, 150.66nm H, L at 132.89nm, H at 52.91nm, L at 85.02nm, H at 54.72nm, 105.86nm L, H at 23.08nm, L at 45.59nm, H at 158.66nm, L at 266.55nm, 137.02nm H, L at 131.61nm, H at 66.09nm, L at 72.64nm, H at 47.04nm, 98.98nm L, 56.18nm H, 25.23nm L, 149.92nm H, 272.53nm L, 125.22nm H, L at 148.06nm, H at 55.02nm, L at 90.84nm, H at 48.47nm, 101.24nm L, 26.63nm H, 118.98nm L, 118.78nm H, 268.06nm L, 113.74nm H, L at 170.23nm, H at 51.04nm, L at 79.74nm, H at 59.02nm, 86.79nm L, 53.31nm H, 75.44nm L, 121.14nm H, 313.65nm L, 101.83nm H, L at 143.17nm, H at 69.3nm, L at 63.27nm, H at 59.67nm, 83.9nm L, 57.83nm H, 78.32nm L, 128.09nm H, 313.49nm L, 92.82nm H, L at 137.42nm, H at 73.12nm, L at 73.84nm, H at 51.33nm, 79.17nm L, 50.88nm H, 100.42nm L, 130.13nm H, 313.34nm L, 77.84nm H, L at 159.16nm, H at 60.01nm, L at 92.87nm, H at 45.03nm, 72.14nm L, 51.53nm H, 118.25nm L, 131.91nm H, 311.73nm L, 53.61nm H, L at 169.66nm, H at 78.72nm, L at 61.79nm, H at 58.2nm, 71.92nm L, H at 69.46nm, L at 41.8nm, H at 171.76nm, L at 279.76nm, 69.79nm H, L at 173.58nm, H at 83.75nm, L at 37.2nm, H at 40.71nm, 87.69nm L, H 60.31nm, L 103.19nm, H 142.79nm, L 299.22nm, 59.62nm H, L at 205.44nm, H at 52.39nm, L at 39.99nm, H at 69.86nm, 28.6nm L, 59.82nm H, 126.8nm L, 58.83nm H, 434.3nm L, 146.52nm The wavelengths are H (124.48 nm), L (12.89 nm), L (41.2 nm), H (73.91 nm), L (48.92 nm), H (191.08 nm), L (426.38 nm), H (11.43 nm), L (214.49 nm), H (8.82 nm), and L (289.74 nm), where H represents a high refractive index material and L represents a low refractive index material.

4. The multi-channel fluorescent dye filter assembly for FISH detection according to claim 1, characterized in that, The film structure of the dichroic filter, from the glass substrate upwards, is as follows: 96.96nm H, 183.69nm L, 124.05nm H, 416.87nm L, 9.46nm H, 31.9nm L, 105.55nm H, 108.11nm L, 16.17nm H, 218.38nm L, 91.76nm H, 78.03nm L, 143.47nm H, 394.33nm L, 20.06nm H, 89.62nm L, 223.41nm H, 171.92nm L, 154.96nm... H, 472.49nm L, 15.38nm H, 246nm L, 66.72nm H, 23.62nm L, 130.21nm H, 238.04nm L, 143nm H, 321.59nm L, 373.71nm H, 49.66nm L, 155.65nm H, 560.02nm L, 57.31nm H, 207.42nm L, 26.2nm H, 123.22nm L, 157.38nm H, 74.79nm L, 134.43nm H, 432.44nm L 103.88nm H, 98.16nm L, 56.59nm H, 99.81nm L, 72nm H, 15.76nm L, 76.3nm H, 223.73nm L, 107.4nm H, 381.78nm L, 94.93nm H, 83.95nm L, 58.92nm H, 104.89nm L, 133.67nm H, 162.16nm L, 86.12nm H, 434.38nm L, 120.4nm H, 172.89nm L, 18.01nm H, 126. The following are the wavelengths of materials: L (93nm), H (188.18nm), L (119.89nm), H (136.13nm), L (334.75nm), H (130.18nm), L (34.33nm), H (91.53nm), L (320.85nm), H (158.68nm), L (529.16nm), H (27.96nm), L (64.82nm), H (41.25nm), L (54.6nm), H (42.05nm), L (236.62nm), H (28.77nm), and L (152.69nm). Here, H represents a high refractive index material, and L represents a low refractive index material.

5. The multi-channel fluorescent dye filter assembly for FISH detection according to claim 1, characterized in that, The center wavelengths of the excitation filter are 435±2nm, 487±2nm, and 565±2nm, and the bandwidths are 20±2nm, 15±2nm, and 20±2nm, respectively. The average transmittance of the excitation filter for light with wavelengths in the range of 425-445nm, 480-494nm, and 555-574nm is >85%, the cutoff range is 452-475nm@OD>6, and the background depth is 500-550nm@OD>6.

6. The multi-channel fluorescent dye filter assembly for FISH detection according to claim 1, characterized in that, The center wavelengths of the emission filter are 462±2nm, 530±2nm, and 605±2nm, with bandwidths of 15±2nm, 20±2nm, and 40±2nm, respectively. The emission filter has an average transmittance of >85% for light with wavelengths in the ranges of 455-470nm, 520-540nm, and 586-624nm, a cutoff range of 475-515nm@OD>6, and a background depth of 547-577nm@OD>6.

7. The multi-channel fluorescent dye filter assembly for FISH detection according to claim 1, characterized in that, The center wavelengths of the dichroic filters are 464±2nm, 530±2nm, and 606±2nm, and their bandwidths are 20±3nm, 20±2nm, and 40±2nm, respectively. The average transmittance of the dichroic filters for light in the wavelength range of 455-472nm, 520-542nm, and 586-626nm is >85%.

8. The multi-channel fluorescent dye filter assembly for FISH detection according to claim 1, characterized in that, The dichroic filter has a reflectivity of >95% for excitation light and a transmittance of >92% for emitted light at an incident angle of 45°.

9. The multi-channel fluorescent dye filter assembly for FISH detection according to any one of claims 2 to 4, characterized in that, The glass substrate is made of borosilicate optical glass, the high refractive index material is tantalum pentoxide, and the low refractive index material is silicon dioxide.

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