Optical system, gene sequencing device, and imaging method

Abstract

Disclosed are an optical system, a gene sequencing device, and an imaging method. The optical system comprises an illumination apparatus and an imaging apparatus; the illumination apparatus comprises a light source and a light guide assembly; the light guide assembly is used for shaping emergent light of the light source to form illumination light; the illumination light is used for being coupled into a gene sequencing chip from a side of the gene sequencing chip and propagating by means of total reflection in the same medium of the gene sequencing chip to generate an evanescent wave; the gene sequencing chip comprises a sample bearing surface that bears a sample; the evanescent wave excites a fluorescent dye in the sample on the sample bearing surface to generate a fluorescence signal; the imaging apparatus is used for generating an imaging light spot on the basis of the fluorescence signal. In this way, the illumination light is coupled into the gene sequencing chip from the side to achieve total internal reflection, only the region of the gene sequencing chip close to the reflective surface is illuminated, the sample near the reflective surface can be excited to produce fluorescence, and free fluorescent molecules distant from the reflective surface cannot be excited to generate background noise, thereby increasing the signal-to-noise ratio of the image.

Description

Optical system, gene sequencing device and imaging method

[0001] The present invention claims priority to Chinese patent application number 202310937995.3, filed with the Patent Office of China on July 28, 2023, entitled “Optical system, gene sequencing device and imaging method”, the entire contents of which are incorporated herein by reference. Technical Field

[0002] The present invention relates to the field of gene sequencing, and in particular to an optical system, a gene sequencing device and an imaging method. Background Art

[0003] When a gene sequencer is working, it needs to use a laser to excite and illuminate the fluorescent dye. The laser light source used is often a Gaussian or super-Gaussian distributed beam, which results in a strong center of the illumination area and a dark edge. Although it can achieve the purpose of illuminating the microscopic object surface, it cannot achieve the optimal lighting effect. In order to excite samples at the edge of the field of view with a sufficient signal-to-noise ratio, the laser power needs to be increased, but at the same time, the laser intensity in the center of the field of view will be too high, resulting in an increased photobleaching rate. In addition, excessive light intensity will damage the DNA in the central field of view, resulting in an increased error rate in subsequent sequencing results.

[0004] Existing gene sequencers often use epi-illumination systems. This, on the one hand, prevents uniform illumination across the object surface, and on the other hand, results in strong background signals, reducing the image signal-to-noise ratio. Therefore, achieving uniform illumination and improving the signal-to-noise ratio remain unresolved technical challenges.

[0005] Summary of the Invention

[0006] The present invention provides an optical system, gene sequencing equipment and imaging method.

[0007] The optical system of the embodiment of the present application includes an illumination device and an imaging device. The illumination device includes a light source and a light guide component. The light guide component is used to shape the output light of the light source to form illumination light. The illumination light is used to couple into the gene sequencing chip from the side and propagate by total reflection within the same medium of the gene sequencing chip to generate an evanescent wave. The gene sequencing chip includes a sample carrying surface carrying a sample. The evanescent wave excites the fluorescent dye in the sample on the sample carrying surface to generate a fluorescent signal. The imaging device is used to generate an imaging spot based on the fluorescent signal.

[0008] The optical system of the embodiment of the present application couples illumination light into the gene sequencing chip from the side to achieve total internal reflection. Only the area of ​​the gene sequencing chip close to the total reflection surface will be illuminated. The sample near the reflection surface can be excited to produce fluorescence, and the free fluorescent molecules far away from the reflection surface will not be excited to produce background noise, thereby greatly increasing the signal-to-noise ratio of the collected image. In addition, this solution can achieve relatively uniform illumination on the surface of the gene sequencing chip, ensuring sequencing efficiency.

[0009] In some embodiments, the gene sequencing chip includes a cover glass, a slide glass and a flow channel, the flow channel is located between the cover glass and the slide glass, the interface between the cover glass and the flow channel is the sample carrying surface, and the illumination light is used to couple into the cover glass from the side of the cover glass, so that the illumination light is totally reflected in the cover glass to illuminate the sample carrying surface.

[0010] In this way, the illumination light can excite the sample located at the interface between the cover glass and the flow channel to generate a fluorescent signal.

[0011] In some embodiments, the gene sequencing chip includes a cover glass, a slide glass and a flow channel, the flow channel is located between the cover glass and the slide glass, the interface between the slide glass and the flow channel is the sample carrying surface, and the illumination light is used to couple into the slide glass from the side of the slide glass, so that the illumination light is totally reflected inside the slide glass to illuminate the sample carrying surface.

[0012] In this way, the illumination light can excite the sample located at the interface between the glass slide and the flow channel to generate a fluorescent signal.

[0013] The gene sequencing chip includes a glass slide, which includes a first surface and a second surface arranged opposite to each other. The sample is carried on the first surface or the second surface to form a sample carrying surface. The illumination light is coupled into the glass slide from the side of the glass slide. The illumination light is totally reflected inside the glass slide to illuminate the sample carrying surface.

[0014] In this way, the illumination light can excite the sample on the slide surface to produce a fluorescent signal.

[0015] In certain embodiments, the gene sequencing chip includes a first side and a second side opposite to the first side, the illumination light is incident into the gene sequencing chip from the first side and is transmitted to the second side, and the illumination device includes a reflective structure arranged on the second side, and the reflective structure is used to reflect the illumination light transmitted to the second side into the gene sequencing chip.

[0016] In this way, the reflective structure can increase the number of times the illumination light is totally reflected in the gene sequencing chip, thereby improving energy utilization.

[0017] In some embodiments, the reflective structure includes at least one of a reflective mirror and a reflective film formed on the second side.

[0018] In this way, the reflective mirror and the reflective film can increase the number of times the illumination light is totally reflected in the gene sequencing chip, thereby improving energy utilization.

[0019] In some embodiments, the light guide assembly includes a collimating lens and a focusing lens. The collimating lens is arranged between the light source and the focusing lens. The collimating lens is used to collimate the output light of the light source, and the focusing lens is used to focus the light passing through the collimating lens to form illumination light.

[0020] In this way, the collimating lens and the focusing lens cooperate with each other to ensure that the illumination light formed after the outgoing light of the light source is collimated and focused has better uniformity, and can make the illumination light coupled into the gene sequencing chip in a concentrated manner.

[0021] In some embodiments, the optical axis of the collimating lens and the optical axis of the focusing lens are coaxially arranged, and the optical axis of the focusing lens is approximately perpendicular to the side of the gene sequencing chip, so that the illumination light can enter the gene sequencing chip vertically from the side of the gene sequencing chip to achieve total reflection.

[0022] In some embodiments, the optical axis of the collimating lens and the optical axis of the focusing lens are coaxially arranged, and the optical axis of the focusing lens is tilted relative to the side of the gene sequencing chip to increase the number of reflections of the illumination light in the gene sequencing chip.

[0023] In this way, through the above technical solution, the incident angle of the illumination light can be increased, and the number of total reflections of the illumination light in the cover glass can be increased, thereby improving energy utilization.

[0024] In some embodiments, the optical axis of the focusing lens is offset relative to the optical axis of the collimating lens to increase the number of reflections of the illumination light within the gene sequencing chip.

[0025] In this way, the incident angle of the illumination light can be increased, and the number of total reflections of the illumination light in the cover glass can be increased, thereby improving energy utilization.

[0026] In certain embodiments, the illumination light is surface illumination light and / or line illumination light.

[0027] In this way, the line illumination light and / or the surface illumination light can change the illumination range of the illumination light on the gene sequencing chip, thereby changing the uniformity of the illumination light.

[0028] The gene sequencing device of the embodiment of the present application includes an optical system, which can improve the efficiency of gene sequencing.

[0029] The imaging method of the embodiment of the present application is to first couple the illumination light emitted by the illumination device into the gene sequencing chip from the side of the gene sequencing chip, so that the illumination light is totally reflected and propagated in the same medium of the gene sequencing chip to generate an evanescent wave. The gene sequencing chip includes a sample-carrying surface carrying a sample. The evanescent wave excites the fluorescent dye in the sample on the sample-carrying surface to generate a fluorescent signal, and then the fluorescent signal is obtained by using an imaging device to generate an imaging light spot.

[0030] In this way, the illumination light is coupled into the gene sequencing chip from the side to achieve total internal reflection. Only the area of ​​the gene sequencing chip close to the total reflection surface will be illuminated. The sample near the reflection surface can be excited to produce fluorescence, and the free fluorescent molecules far away from the reflection surface will not be excited to produce background noise, thereby greatly increasing the signal-to-noise ratio of the collected image. In addition, this solution can achieve better uniformity of illumination on the surface of the gene sequencing chip, ensuring sequencing efficiency.

[0031] Additional aspects and advantages of the present invention will be set forth in part in the description which follows and, in part, will be obvious from the description which follows, or may be learned by practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] The above and / or additional aspects and advantages of the present invention will become apparent and readily understood from the description of the embodiments with reference to the following drawings, in which:

[0033] FIG1 is a schematic structural diagram of an optical system according to an embodiment of the present invention;

[0034] FIG2 is a schematic diagram of the structure of a gene sequencing chip according to an embodiment of the present invention;

[0035] FIG3 is a schematic diagram of the cooperation between an illumination device and a gene sequencing chip according to one embodiment of the present invention;

[0036] FIG4 is a distribution diagram of energy density of illumination light on a gene sequencing chip according to one embodiment of the present invention;

[0037] FIG5 is a schematic structural diagram of a gene sequencing chip according to an embodiment of the present invention;

[0038] FIG6 is a schematic diagram of the cooperation between an illumination device and a gene sequencing chip according to one embodiment of the present invention;

[0039] FIG7 is a distribution diagram of energy density of illumination light on a gene sequencing chip according to one embodiment of the present invention;

[0040] FIG8 is a schematic structural diagram of a gene sequencing chip according to an embodiment of the present invention;

[0041] FIG9 is a schematic diagram of the cooperation between an illumination device and a gene sequencing chip according to another embodiment of the present invention;

[0042] FIG10 is a diagram showing the energy density distribution of illumination light on a gene sequencing chip according to another embodiment of the present invention;

[0043] FIG11 is a schematic diagram of the cooperation between the lighting device and the gene sequencing chip according to an embodiment of the present invention;

[0044] FIG12 is a schematic diagram of the cooperation between the lighting device and the gene sequencing chip according to an embodiment of the present invention;

[0045] FIG13 is a schematic diagram of the cooperation between an illumination device and a gene sequencing chip according to another embodiment of the present invention;

[0046] FIG14 is a diagram showing the energy density distribution of illumination light on a gene sequencing chip according to another embodiment of the present invention;

[0047] FIG15 is a schematic diagram of the cooperation between an illumination device and a gene sequencing chip according to yet another embodiment of the present invention;

[0048] FIG16 is a diagram showing the energy density distribution of illumination light on a gene sequencing chip according to yet another embodiment of the present invention;

[0049] FIG17 is a schematic diagram of a linear illumination light according to an embodiment of the present invention;

[0050] FIG18 is a schematic diagram of surface illumination according to an embodiment of the present invention;

[0051] FIG19 is a diagram showing the energy density distribution of surface illumination light on a gene sequencing chip according to an embodiment of the present invention;

[0052] FIG20 is a flow chart of an imaging method of an optical system according to an embodiment of the present invention.

[0053] Description of reference numerals:

[0054] 100. Optical system; 10. Illuminating device; 11. Light source; 111. Illuminating light; 12. Light guide assembly; 121. Collimating lens; 122. Focusing lens; 13. Reflective structure; 20. Imaging device; 200. Gene sequencing chip; 201. Cover glass; 202. Slide; 2021. First surface; 2022. Second surface; 203. Flow channel; 204. Sample holding surface; 205. Sample; 206. First side; 207. Second side. DETAILED DESCRIPTION

[0055] The embodiments of the present invention are described in detail below, examples of which are shown in the accompanying drawings, wherein the same or similar reference numerals throughout represent the same or similar elements or elements having the same or similar functions. The embodiments described below with reference to the accompanying drawings are exemplary and are only used to explain the present invention, and are not to be construed as limiting the present invention.

[0056] In the description of the present invention, it should be understood that the terms "center", "longitudinal", "lateral", "length", "width", "thickness", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside", "clockwise", "counterclockwise" and the like, indicating orientations or positional relationships, are based on the orientations or positional relationships shown in the accompanying drawings, and are only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as limiting the present invention. In addition, the terms "first" and "second" are used for descriptive purposes only, and cannot be understood as indicating or implying relative importance or implicitly indicating the number of the indicated technical features. Therefore, the features defined as "first" and "second" may explicitly or implicitly include one or more of the said features. In the description of the present invention, the meaning of "multiple" is two or more, unless otherwise clearly and specifically defined.

[0057] In the description of the present invention, it should be noted that, unless otherwise expressly specified or limited, the terms "mounted," "connected," and "connected" should be understood in a broad sense. For example, they may refer to fixed connections, detachable connections, or integral connections; mechanical connections, electrical connections, or mutual communication; direct connections or indirect connections through an intermediate medium; and internal communication between two components or interaction between two components. Those skilled in the art will understand the specific meanings of the above terms in the present invention based on specific circumstances.

[0058] In the present invention, unless otherwise expressly specified or limited, a first feature being "above" or "below" a second feature may include the first and second features being in direct contact, or may include the first and second features being in contact not directly but through another feature between them. Furthermore, a first feature being "above," "above," and "above" a second feature may include the first feature being directly above or obliquely above the second feature, or may simply mean that the first feature is higher in level than the second feature. A first feature being "below," "below," and "below" a second feature may include the first feature being directly below or obliquely below the second feature, or may simply mean that the first feature is lower in level than the second feature.

[0059] The disclosure below provides many different embodiments or examples for realizing different structures of the present invention. In order to simplify the disclosure of the present invention, the components and settings of specific examples are described below. Of course, they are merely examples and are not intended to limit the present invention. In addition, the present invention may repeat reference numbers and / or reference letters in different examples. Such repetition is for the purpose of simplicity and clarity and does not in itself indicate the relationship between the various embodiments and / or settings discussed. In addition, the present invention provides examples of various specific processes and materials, but those skilled in the art will recognize the application of other processes and / or the use of other materials.

[0060] Referring to Figures 1 and 2, the optical system 100 of the embodiment of the present application includes an illumination device 10 and an imaging device 20. The illumination device 10 includes a light source 11 and a light guide component 12. The light guide component 12 is used to shape the output light of the light source 11 to form illumination light 111. The illumination light 111 is used to couple into the gene sequencing chip 200 from the side and propagate in the same medium of the gene sequencing chip 200 by total reflection to generate an evanescent wave. The gene sequencing chip 200 includes a sample carrying surface 204 carrying a sample 205. The evanescent wave excites the fluorescent dye in the sample 205 on the sample carrying surface 204 to generate a fluorescent signal; the imaging device 20 is used to generate an imaging spot based on the fluorescent signal.

[0061] The optical system 100 of the embodiment of the present application couples the illumination light 111 from the side of the gene sequencing chip 200 into the gene sequencing chip 200 to achieve total internal reflection. Only the area of ​​the gene sequencing chip 200 close to the total reflection surface will be illuminated, and the sample 205 near the reflection surface can be excited to produce fluorescence. The free fluorescent molecules far from the reflection surface will not be excited to produce background noise, thereby greatly increasing the signal-to-noise ratio of the collected image. In addition, this solution can achieve relatively uniform illumination on the surface of the gene sequencing chip 200, ensuring sequencing efficiency.

[0062] Specifically, the optical system 100 is a system composed of one or more optical elements with the required optical function, and the optical system 100 can produce a clear image that is completely similar to the appearance of the object. The imaging device 20 is a device that converts optical signals into visual images.

[0063] The light guide assembly 12 is used to shape the light emitted by the light source 11. This means that the light guide assembly 12 can adjust parameters such as the shape, direction, and illumination range of the light emitted by the light source 11. The illumination light 111 is coupled into the gene sequencing chip 200 from the side. This means that the illumination light 111 can pass through the side of the gene sequencing chip 200 and enter the gene sequencing chip 200.

[0064] In the embodiment of the present invention, the same medium is an object with a consistent refractive index. For example, the object can be made of a material with uniform density, so that the refractive index at each position of the object is consistent.

[0065] An evanescent wave is an electromagnetic wave generated on one side of an optically less dense medium when light is obliquely incident on an optically less dense medium, resulting in total internal reflection. This electromagnetic wave can excite sample 205 to produce a fluorescent signal. Because the evanescent wave generated by total internal reflection is only a few hundred nanometers thick and cannot propagate deeply within gene sequencing chip 200, the background of gene sequencing chip 200 is not illuminated, allowing imaging device 20 to capture images with a high signal-to-noise ratio.

[0066] The sample holding surface 204 is used to hold the sample 205. In some embodiments, the sample 205 may be a base cluster with a fluorescent dye. The wavelength of the light emitted by the light source 11 may be 532 nm, the optical fiber aperture may be 0.2*0.2 mm, and the numerical aperture NA may be 0.22.

[0067] Referring to Figures 2 and 3, in some embodiments, the gene sequencing chip 200 includes a cover glass 201, a slide glass 202 and a flow channel 203. The flow channel 203 is located between the cover glass 201 and the slide glass 202. The interface between the cover glass 201 and the flow channel 203 is a sample carrying surface 204. The illumination light 111 is used to couple into the cover glass 201 from the side of the cover glass 201, so that the illumination light 111 is totally reflected in the cover glass 201 to illuminate the sample carrying surface 204.

[0068] In this way, the illumination light 111 can excite the sample 205 located at the interface between the cover glass 201 and the flow channel 203 to generate a fluorescent signal.

[0069] Specifically, the illumination light 111 is coupled into the cover glass 201 from the side of the cover glass 201. The illumination light 111 undergoes multiple total reflections inside the cover glass 201, illuminating the sample 205 located at the interface between the cover glass 201 and the flow channel 203. The fluorescent dye on the sample 205 is excited by the illumination light 111, generating a fluorescence signal. The imaging device 20 generates an imaging spot based on the fluorescence signal.

[0070] In some embodiments, the thickness of the cover glass 201 can be 170 μm, the refractive index of the cover glass 201 can be 1.52, the thickness of the flow channel 203 can be 80 μm, and the refractive index of the liquid in the flow channel 203 can be 1.33. Therefore, the refractive index of the cover glass 201 is greater than the refractive index of the liquid in the flow channel 203, so that the illumination light 111 can be totally reflected in the cover glass 201.

[0071] Please refer to Figure 4 . In Figure 4 , the abscissa represents various locations along the short side of gene sequencing chip 200, and the ordinate represents the energy density of illumination light 111. As shown in Figure 4 , within the range of -12.5 mm to 8 mm on the abscissa, the energy density curve of illumination light 111 demonstrates a uniformity exceeding 95%, with an energy density of approximately 16 W / cm².

[0072] Exemplarily, the uniformity of the illumination light 111 may be calculated using the following formula: 1-(max-min) / (max+min), where max is the maximum value of the energy density within the calculation range, and min is the minimum value of the energy density within the calculation range.

[0073] Referring to Figures 5 and 6, in some embodiments, the gene sequencing chip 200 includes a cover glass 201, a glass slide 202 and a flow channel 203. The flow channel 203 is located between the cover glass 201 and the glass slide 202. The interface between the glass slide 202 and the flow channel 203 is a sample carrying surface 204. The illumination light 111 is used to couple into the glass slide 202 from the side of the glass slide 202, so that the illumination light 111 is totally reflected in the glass slide 202 to illuminate the sample carrying surface 204.

[0074] In this way, the illumination light 111 can excite the sample 205 located at the interface between the glass slide 202 and the flow channel 203 to generate a fluorescent signal.

[0075] Specifically, the illumination light 111 is coupled into the glass slide 202 from the side of the glass slide 202, and the illumination light 111 undergoes multiple total reflections within the glass slide 202, illuminating the sample 205 located at the interface between the glass slide 202 and the flow channel 203. The fluorescent dye on the gene sequencing chip 200 is excited by the illumination light 111, generating a fluorescence signal, and the imaging device 20 generates an imaging spot based on the fluorescence signal.

[0076] In some embodiments, the thickness of the flow channel 203 can be 80 μm, the refractive index of the liquid in the flow channel 203 can be 1.33, and the thickness of the glass slide 202 can be 1 mm, and the refractive index of the glass slide 202 can be 1.52. Therefore, the refractive index of the glass slide 202 is greater than the refractive index of the liquid in the flow channel 203, so that the illumination light 111 can be totally reflected within the glass slide 202.

[0077] Please refer to Figure 7 . In Figure 7 , the horizontal axis represents various positions along the length of gene sequencing chip 200, and the vertical axis represents the energy density of illumination light 111. As shown in Figure 7 , within the range of -12.5 mm to 8 mm on the horizontal axis, the energy density curve of illumination light 111 shows a uniformity exceeding 90%, with an energy density of approximately 150 W / cm².

[0078] Exemplarily, the uniformity of the illumination light 111 may be calculated using the following formula: 1-(max-min) / (max+min), where max is the maximum value of the energy density within the calculation range, and min is the minimum value of the energy density within the calculation range.

[0079] Referring to Figures 1 and 8, in some embodiments, the gene sequencing chip 200 includes a glass slide 202, and the glass slide 202 includes a first surface 2021 and a second surface 2022 arranged opposite to each other. The sample 205 is carried on the first surface 2021 or the second surface 2022 to form a sample carrying surface 204. The illumination light 111 is coupled into the glass slide 202 from the side of the glass slide 202, and the illumination light 111 is totally reflected in the glass slide 202 to illuminate the sample carrying surface 204.

[0080] In this way, the illumination light 111 can excite the sample 205 located on the surface of the glass slide 202 to produce a fluorescence signal. Specifically, the illumination light 111 is coupled into the glass slide 202 from the side. The illumination light 111 undergoes multiple total reflections within the glass slide 202, illuminating the sample 205 located on the surface of the glass slide 202. The fluorescent dye on the sample 205 is excited by the illumination light 111, producing a fluorescence signal. The imaging device 20 generates an imaging spot based on the fluorescence signal. As shown in Figure 8, the sample 205 is supported on the first surface 2021, which in this case forms the sample supporting surface 204. In other embodiments, the sample 205 can be supported on the second surface 2022, which forms the sample supporting surface 204.

[0081] Referring to Figures 1, 5 and 9, in some embodiments, the gene sequencing chip 200 includes a first side 206 and a second side 207 opposite to the first side 206. The illumination light 111 is incident into the gene sequencing chip 200 from the first side 206 and is transmitted to the second side 207. The illumination device 10 includes a reflective structure 13 arranged on the second side 207. The reflective structure 13 is used to reflect the illumination light 111 transmitted to the second side 207 into the gene sequencing chip 200.

[0082] In this way, the reflective structure 13 can increase the number of total reflections of the illumination light 111 within the gene sequencing chip 200, thereby improving energy utilization.

[0083] In some embodiments, the reflective structure 13 includes at least one of a reflective mirror and a reflective film formed on the second side surface 207 .

[0084] In this way, the reflective mirror and the reflective film can increase the number of times the illumination light 111 is totally reflected in the gene sequencing chip 200, thereby improving energy utilization.

[0085] Specifically, in some embodiments, the illumination light 111 is coupled into the cover glass 201 from the first side 206 of the cover glass 201 , a reflector can be added to the second side 207 of the cover glass 201 , or a reflective film can be coated on the second side 207 of the cover glass 201 .

[0086] Please refer to Figure 10. In Figure 10, the abscissa represents various positions along the length of gene sequencing chip 200, and the ordinate represents the energy density of illumination light 111. As shown in Figure 10, within the abscissa range of -12.5 mm to 8 mm, the energy density curve of illumination light 111 achieves a uniformity exceeding 95%, with an energy density of approximately 32 W / cm². Compared to the energy density of approximately 16 W / cm² in Figure 4, this represents a roughly doubled increase in energy efficiency.

[0087] Please refer to Figures 11 and 12. In some embodiments, the light guide component 12 includes a collimating lens 121 and a focusing lens 122. The collimating lens 121 is arranged between the light source 11 and the focusing lens 122. The collimating lens 121 is used to collimate the output light of the light source 11, and the focusing lens 122 is used to converge the light passing through the collimating lens 121 to form the illumination light 111.

[0088] In this way, the collimating lens 121 and the focusing lens 122 cooperate with each other to ensure that the illumination light 111 formed after the outgoing light of the light source 11 is collimated and converged has better uniformity, and the illumination light 111 can be concentratedly coupled into the gene sequencing chip 200.

[0089] Specifically, the focal length of the collimating lens 121 may be 2 mm to 200 mm, the focal length of the condensing lens 122 may be 4 mm to 50 mm, the collimating lens 121 is a spherical lens, and the condensing lens 122 is a cylindrical lens.

[0090] Please refer to Figure 9. In some embodiments, the optical axis of the collimating lens 121 and the optical axis of the focusing lens 122 are coaxially arranged, and the optical axis of the focusing lens 122 is approximately perpendicular to the side of the gene sequencing chip 200, so that the illumination light 111 can enter the gene sequencing chip 200 vertically from the side of the gene sequencing chip 200 to achieve total reflection.

[0091] Specifically, the optical axis of the collimating lens 121 is the central axis of the collimating lens 121 , and the optical axis of the condensing lens 122 is the central axis of the condensing lens 122 .

[0092] Please refer to Figure 13. In some embodiments, the optical axis of the collimating lens 121 and the optical axis of the focusing lens 122 are coaxially arranged, and the optical axis of the focusing lens 122 is tilted relative to the side of the gene sequencing chip 200 to increase the number of reflections of the illumination light 111 in the gene sequencing chip 200.

[0093] In this way, through the above technical solution, the incident angle of the illumination light 111 can be increased, thereby increasing the number of total reflections of the illumination light 111 in the cover glass 201, thereby improving energy utilization.

[0094] Specifically, the angle between the optical axis of the condenser lens 122 and the side of the gene sequencing chip 200 can be greater than the critical angle at which no refraction occurs. The closer the incident angle of the illumination light 111 is to the critical angle, the more times the illumination light 111 is totally reflected within the cover glass 201. In some embodiments, the light source 11 and light guide assembly 12 can be tilted 10° relative to the gene sequencing chip 200, thereby increasing the energy utilization efficiency of the illumination light 111 by approximately five times.

[0095] Please refer to Figure 14 . In Figure 14 , the abscissa represents various positions along the length of gene sequencing chip 200, and the ordinate represents the energy density of illumination light 111. As shown in Figure 14 , within the abscissa range of -12.5 mm to 8 mm, the energy density curve of illumination light 111 achieves a uniformity exceeding 95%, with an energy density of approximately 160 W / cm². Compared to the energy density of approximately 32 W / cm² in Figure 10 , this represents a fivefold improvement in energy efficiency.

[0096] Referring to FIG. 15 , in some embodiments, the optical axis of the condensing lens 122 is offset relative to the optical axis of the collimating lens 121 to increase the number of reflections of the illumination light 111 within the gene sequencing chip 200 .

[0097] In this way, the incident angle of the illumination light 111 can be increased, and the number of total reflections of the illumination light 111 in the cover glass 201 can be increased, thereby improving energy utilization.

[0098] Specifically, in some embodiments, the optical axis of the condenser lens 122 is offset by 0.7 mm relative to the optical axis of the collimating lens 121, thereby increasing energy utilization by approximately 5.3 times. Referring to Figure 16 , the abscissa represents various positions along the length of the gene sequencing chip 200, and the ordinate represents the energy density of the illumination light 111. As can be seen from Figure 16 , within the abscissa range of -12.5 mm to 8 mm, the energy density of the illumination light 111 achieves a uniformity exceeding 95%, with an energy density of approximately 168 W / cm². This represents a 5.3-fold increase in energy utilization compared to the approximately 32 W / cm² shown in Figure 10 .

[0099] 17 and 18 , in some embodiments, the illumination light 111 is a linear illumination light 111 and / or a surface illumination light 111 .

[0100] In this way, the line illumination light 111 and / or the surface illumination light 111 can change the illumination range of the illumination light 111 on the gene sequencing chip 200 , thereby changing the uniformity of the illumination light 111 .

[0101] Specifically, in some embodiments, when the focal length of the collimating lens 121 is 3 mm and the focal length of the focusing lens 122 is 4 mm, the illumination light 111 is line illumination light 111, and the uniformity of the line illumination light 111 is more than 95%; please refer to Figure 19, when the focal length of the collimating lens 121 is 75 mm and the focal length of the focusing lens 122 is 50 mm, the illumination light 111 is surface illumination light 111, and the uniformity of the surface illumination light 111 is more than 85%.

[0102] The gene sequencing device according to the embodiment of the present application includes an optical system 100. In this way, the efficiency of gene sequencing can be improved.

[0103] Referring to FIG. 20 , the imaging method according to the embodiment of the present application includes the following steps:

[0104] S10, coupling the illumination light 111 emitted by the illumination device 10 into the gene sequencing chip 200 from the side surface thereof, so that the illumination light 111 is totally reflected and propagates within the same medium of the gene sequencing chip 200 to generate an evanescent wave. The gene sequencing chip 200 includes a sample carrying surface 204 carrying a sample 205. The evanescent wave excites a fluorescent dye in the sample 205 on the sample carrying surface 204 to generate a fluorescent signal.

[0105] S20 , using the imaging device 20 to acquire a fluorescent signal to generate an imaging spot.

[0106] In this way, the illumination light 111 is coupled into the gene sequencing chip 200 from the side to achieve total internal reflection. Only the area of ​​the gene sequencing chip 200 close to the total reflection surface will be illuminated, and the sample 205 near the reflection surface can be excited to produce fluorescence. The free fluorescent molecules far away from the reflection surface will not be excited to produce background noise, thereby greatly increasing the signal-to-noise ratio of the collected image. In addition, this solution can achieve relatively uniform illumination on the surface of the gene sequencing chip 200, ensuring sequencing efficiency.

[0107] Throughout this specification, reference to terms such as "one embodiment," "certain embodiments," "illustrative embodiments," "examples," "specific examples," or "some examples" means that a specific feature, structure, material, or characteristic described in conjunction with the embodiment or example is included in at least one embodiment or example of the present invention. In this specification, illustrative descriptions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.

[0108] While embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions, and variations may be made to the embodiments without departing from the principles and spirit of the invention, and that the scope of the invention is defined by the claims and their equivalents.

Claims

1. An optical system, characterized in that: include: An illumination device, comprising a light source and a light guide component, wherein the light guide component is used to shape the emitted light of the light source to form illumination light, wherein the illumination light is used to couple into the gene sequencing chip from the side thereof and propagate by total reflection in the same medium of the gene sequencing chip to generate an evanescent wave, wherein the gene sequencing chip comprises a sample carrying surface carrying a sample, wherein the evanescent wave excites a fluorescent dye in the sample on the sample carrying surface to generate a fluorescent signal; and, An imaging device is used to generate an imaging spot according to the fluorescent signal.

2. The optical system according to claim 1, characterized in that The gene sequencing chip includes a cover glass, a slide glass and a flow channel, wherein the flow channel is located between the cover glass and the slide glass, and the interface between the cover glass and the flow channel is the sample carrying surface. The illumination light is used to couple into the cover glass from the side of the cover glass, so that the illumination light is totally reflected in the cover glass to illuminate the sample carrying surface.

3. The optical system according to claim 1, characterized in that The gene sequencing chip includes a cover glass, a glass slide and a flow channel, wherein the flow channel is located between the cover glass and the glass slide, and the interface between the glass slide and the flow channel is the sample carrying surface. The illumination light is used to couple into the glass slide from the side of the glass slide, so that the illumination light is totally reflected in the glass slide to illuminate the sample carrying surface.

4. The optical system according to claim 1, characterized in that The gene sequencing chip includes a glass slide, which includes a first surface and a second surface arranged opposite to each other. The sample is carried on the first surface or the second surface to form the sample carrying surface. The illumination light is coupled into the glass slide from the side of the glass slide. The illumination light is totally reflected in the glass slide to illuminate the sample carrying surface.

5. The optical system according to claim 1, characterized in that The gene sequencing chip includes a first side and a second side opposite to the first side. The illumination light is incident into the gene sequencing chip from the first side and is transmitted to the second side. The illumination device includes a reflection structure arranged on the second side. The reflection structure is used to reflect the illumination light transmitted to the second side into the gene sequencing chip.

6. The optical system according to claim 5, characterized in that The reflective structure includes at least one of a reflective mirror and a reflective film formed on the second side surface.

7. The optical system according to claim 1, characterized in that The light guide assembly includes a collimating lens and a condensing lens. The collimating lens is arranged between the light source and the condensing lens. The collimating lens is used to collimate the outgoing light of the light source, and the condensing lens is used to converge the light passing through the collimating lens to form illumination light.

8. The optical system according to claim 7, characterized in that The optical axis of the collimating lens is coaxially arranged with the optical axis of the condensing lens, and the optical axis of the condensing lens is perpendicular to the side surface of the gene sequencing chip.

9. The optical system according to claim 7, characterized in that The optical axis of the collimating lens and the optical axis of the condensing lens are coaxially arranged, and the optical axis of the condensing lens is inclined relative to the side of the gene sequencing chip to increase the number of reflections of the illumination light in the gene sequencing chip.

10. The optical system according to claim 7, characterized in that The optical axis of the condensing lens is offset relative to the optical axis of the collimating lens to increase the number of reflections of the illumination light in the gene sequencing chip.

11. The optical system according to claim 1, characterized in that The illumination light is line illumination light and / or surface illumination light.

12. A gene sequencing device, characterized in that: An optical system comprising any one of claims 1-11.

13. An imaging method, characterized in that: include: The illumination light emitted by the illumination device is coupled into the gene sequencing chip from the side of the gene sequencing chip, so that the illumination light is totally reflected and propagated in the same medium of the gene sequencing chip and generates an evanescent wave, wherein the gene sequencing chip includes a sample carrying surface carrying a sample, and the evanescent wave excites a fluorescent dye in the sample on the sample carrying surface to generate a fluorescent signal; The fluorescence signal is acquired by an imaging device to generate an imaging spot.

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