Gene sequencing system and method provided with multi-surface aberration compensator

Abstract

A gene sequencing system and method provided with a multi-surface aberration compensator, which system and method are used for imaging a fluorescence signal of a sequencing chip. An inherent aberration adjustment mirror is disposed in an imaging system, such that the use of an insertion-type adjustor is avoided, thereby reducing the problems of precision and redundancy caused by insertion; and aberration generated within a certain flow channel thickness (or chip thickness) range can be continuously adjusted, thereby improving the adaptability of the range of application, and reducing costs. By means of adjusting the aberration adjustment mirror, imaging of a first surface and a second surface of flow channels of different chips can be realized without needing to replace an optical element in the imaging system, such that a wide range of applicability is achieved.

Description

Gene sequencing system and method provided with multi-surface aberration compensator Technical Field

[0001] The present invention relates to the field of biometric imaging technology, and in particular to a gene sequencing system and method provided with a multi-surface aberration compensator. Background Art

[0002] Due to its optical precision and wide applicability, microscopy technology can be used to detect fluorescence-excited biosample chip signals in gene sequencing. In gene sequencing technology, a biochip is provided with a flow channel, which includes a first surface (the inner surface located above the flow channel) and a second surface (the inner surface located below the flow channel). To improve chip utilization and detection throughput, both the first and second surfaces of the flow channel are generally configured as detection surfaces, and an optical microscope is used to detect the fluorescence signals generated by the excitation of the biological sample on the first and second surfaces.

[0003] The upper and lower surfaces are tested, and the thickness of the flow channel is about 20-200 microns. The objective lens used in the gene sequencer has a large NA (numerical aperture) and a small depth of field. The thickness of the flow channel and the fluid medium in the flow channel will have a great aberration effect on the imaging quality. The current industry adopts the method of inserting optical correction devices to compensate for the aberration caused by the thickness of the flow channel, so that the imaging system can clearly image the upper and lower surfaces of the biological sample chip. However, the insertion method changes the number of optical elements in the optical imaging system, and there will be problems with reduced insertion accuracy and repeatability. The inserted regulator only compensates for the aberration caused by a certain thickness of the flow channel (that is, the aberration compensation value is fixed). At the same time, multiple repetitive operations also increase the complexity of the operation, and its applicability is relatively limited.

[0004] Existing imaging systems have poor applicability for imaging high-throughput sequencing systems, poor stability and repeatability, and high costs.

[0005] Summary of the Invention

[0006] In view of this, the object of the present invention is to provide a gene sequencing system and method provided with a multi-surface aberration compensator, which uses an aberration adjustment mirror to adjust the imaging aberration during biological measurement to improve imaging accuracy.

[0007] In order to achieve the above object, the present invention provides the following technical solutions:

[0008] The gene sequencing system provided by the present invention and provided with a multi-surface aberration compensator comprises a gene sequencer and a multi-surface aberration compensator;

[0009] The multi-surface aberration compensator includes at least one aberration adjustment mirror for compensating for aberrations generated by a signal beam of a gene sequencer during an imaging process;

[0010] The light beam emitted from the incident light path in the gene sequencer is irradiated onto biological samples located on different surfaces through the objective lens of the gene sequencer. The biological samples are excited by the light beam to generate signal light beams. The signal light beams reflected back from different surfaces are focused by the corresponding aberration adjustment mirrors and then enter the imaging unit to generate images.

[0011] Furthermore, it also includes a control module for controlling the aberration adjustment mirror, which is used to obtain the aberration compensation adjustment value of the aberration generated by different surface spacings and adjust the focal length of the multi-surface aberration compensator according to the aberration compensation adjustment value.

[0012] Furthermore, the different surfaces are the first surface and the second surface of the biochip flow channel arranged on the upper gene sequencer; the first surface is the upper surface of the flow channel in the biochip, and the second surface is the lower surface of the flow channel in the biochip.

[0013] Furthermore, the radiation light beam emitted by the incident light path enters the first surface of the flow channel of the biochip through the objective lens. In the first control state of the aberration adjustment mirror, the signal light beam reflected back from the first surface of the flow channel passes through the aberration adjustment mirror and enters the imaging unit for imaging; when the focal length of the aberration adjustment mirror is changed, in the second control state of the aberration adjustment mirror, the light beam signal reflected back from the second surface of the flow channel passes through the aberration adjustment mirror and enters the imaging unit for imaging.

[0014] Furthermore, the imaging unit includes a reflective light path splitter, a first tube lens, a first photosensitive device unit, a second tube lens, and a second photosensitive device unit;

[0015] The reflective light path splitter is used to split the light beam reflected from the flow channel collected by the objective lens into a first light beam and a second light beam. The first light beam enters the first tube lens and converges to be received and imaged by the first photosensitive device unit. The second light beam enters the second tube lens and converges to be received and imaged by the second photosensitive device unit.

[0016] Furthermore, the imaging unit further comprises a first beam splitter, a third tube lens, and a third photosensitive device unit; a second beam splitter, a fourth tube lens, and a fourth photosensitive device unit;

[0017] The first light beam enters the first beam splitter and is divided into a third light beam and a fourth light beam. The third light beam enters the first tube lens and converges to the first photosensitive device unit for receiving imaging. The fourth light beam enters the third tube lens and converges to the third photosensitive device unit for receiving imaging. At the same time,

[0018] After entering the second beam splitter, the second beam is divided into a fifth beam and a sixth beam. After entering the second tube lens, the fifth beam converges to the second photosensitive device unit for receiving imaging. After entering the fourth tube lens, the sixth beam converges to the fourth photosensitive device unit for receiving imaging.

[0019] Furthermore, the aberration adjustment mirror includes a film lens, a container, and a driver;

[0020] The film lens is arranged on the container, and an optical fluid material is arranged in the closed chamber formed between the film lens and the container. The driver applies pressure to the container and causes the film lens to deform so as to change the focal length of the imaging module composed of the film lens, the optical fluid material and the container.

[0021] Furthermore, the film lens in the aberration adjustment mirror is a deformable lens, which is made of elastic polymer. The film lens is sealed on a container filled with liquid medium, and a driver is used to apply pressure to the container, causing the film lens to bend so that the focal length changes.

[0022] Furthermore, the driver controls the pressure applied to the container through a control circuit, and the adjustment current of the driver is in a linear relationship with the focal length of the lens.

[0023] Furthermore, the optical fluid material is selected from a material with a refractive index of about 1.1-1.5 and an Abbe number of 90-110.

[0024] Furthermore, the optical fluid material is a low dispersion material with a refractive index of 1.3 and an Abbe number of 100.

[0025] Furthermore, the driver includes a voice coil motor and a controller. The voice coil motor, controller, thin film lens, and container constitute a voice coil liquid lens. The voice coil motor generates a driving force on the container under the action of the controller, so that the focal length of the voice coil liquid lens changes.

[0026] Furthermore, the voice coil motor includes a mover component and a stator component; the mover component and the stator component constitute an electromagnetic drive device;

[0027] The axis of the movable component coincides with that of the stator component, and both the movable component and the stator component are electrically connected to a controller; under the control of the controller, the movable component and the stator component work, and under the action of the electromagnetic field, the movable component and the stator component move, thereby providing a driving force for linear motion.

[0028] Furthermore, the stator component is arranged on the container, and the mover component is arranged on the film lens. The mover component is used to apply a driving force to the film lens, so that the film lens is deformed.

[0029] Furthermore, the movable component adopts either a coil or a magnet, and the stator component adopts either a coil or a magnet. When the movable component adopts a coil, the stator component is a magnet; when the movable component adopts a magnet, the stator component is a coil; so that the movable component and the stator component can constitute an electromagnetic drive device that can provide electromagnetic force.

[0030] Furthermore, the electromagnetic driving device adopts two types: moving coil type and moving iron type, and generates different driving forces by changing the coil current, and changes the focal length value of the liquid lens under the action of the driving force.

[0031] Furthermore, the film lens adopts a non-uniform thickness elastic film structure, the light-transmitting area of ​​the film lens is arranged as a convex-flat structure, and the refractive index of the liquid filled in the liquid cavity matches the refractive index of the non-uniform thickness elastic film.

[0032] Furthermore, the movable component adopts a driving ring arranged on the film lens, which drives the film lens in the light-transmitting area in the middle of the ring to deform by squeezing or stretching the driving ring.

[0033] Furthermore, the movable component is adhered to the driving ring through liquid. By controlling the magnitude and direction of the coil current, the liquid lens can be driven to produce deformations in different directions and degrees. The curvature of the thin film lens is changed by the voice coil motor, thereby changing the focal length of the lens.

[0034] The method for implementing gene sequencing using the above-mentioned gene sequencing system provided by the present invention comprises the following steps:

[0035] placing a biological sample to be tested in the flow channel of the biochip on the carrier;

[0036] Adjusting the focal length of the aberration adjustment mirror to a first state so that a signal light beam generated by the biological sample disposed on the first surface and excited by the radiation light source passes through the first state and is imaged by the imaging unit;

[0037] Re-adjusting the focal length of the aberration adjustment mirror to a second state so that the signal light beam generated by the biological sample disposed on the second surface and excited by the radiation light source passes through the second state and is imaged by the imaging unit;

[0038] Repeat the above steps so that the signal light beams generated by the biological samples on all surfaces are excited by the radiation light source and then focused by the aberration adjustment mirror and imaged by the imaging unit;

[0039] The gene sequencing results are obtained through a gene sequencer based on the imaging data.

[0040] Furthermore, the signal light beam passes through a spectroscope to obtain two light beam signals, and the two light beam signals are respectively imaged in corresponding photosensitive device units.

[0041] Furthermore, the signal light beam is passed through a spectroscope to obtain two light beam signals, which are then split again through a spectroscope to obtain four light beam signals, and each light beam signal is imaged in a corresponding photosensitive device unit.

[0042] The method for implementing gene sequencing using the above-mentioned gene sequencing system provided by the present invention comprises the following steps:

[0043] acquiring position data of different surfaces on which biological samples are placed;

[0044] Calculate compensation adjustment values ​​for generating aberrations based on position data of different surfaces;

[0045] The aberration adjustment mirror is adjusted according to the compensation adjustment value of the aberration to obtain imaging data of the signal light beam generated by the biological sample on different surfaces excited by the radiation light source in the imaging unit;

[0046] The gene sequencing results are obtained through a gene sequencer based on the imaging data.

[0047] Furthermore, the different surfaces of the biological sample are a first surface and a second surface of a biochip flow channel;

[0048] The position data of the different surfaces include a first distance value and a second distance value;

[0049] The first distance value is the distance between the signal light beam generated by the biological sample excited by the radiation light source on the first surface of the flow channel and the imaging unit when imaging;

[0050] The second distance value is a distance between the signal light beam generated by the biological sample excited by the radiation light source and the imaging unit when imaging.

[0051] The beneficial effects of the present invention are:

[0052] The present invention provides a gene sequencing system equipped with a multi-surface aberration compensator for imaging fluorescence signals from sequencing chips. By providing an inherent aberration adjustment mirror within the imaging system, the use of an insertable adjuster is avoided, reducing accuracy and repeatability issues caused by insertion. Furthermore, the system can continuously adjust aberrations generated within a certain range of flow channel thickness (or chip thickness), thereby improving adaptability and reducing costs. Adjusting the aberration adjustment mirror allows imaging of the first and second surfaces of flow channels on different chips without requiring replacement of optical components in the imaging system, thus achieving wide applicability.

[0053] Other advantages, objects, and features of the present invention will be described in part in the following description and, in part, will be apparent to those skilled in the art upon examination of the following description or may be learned from practice of the present invention. The objects and other advantages of the present invention may be realized and obtained through the following description. BRIEF DESCRIPTION OF THE DRAWINGS

[0054] In order to make the purpose, technical solutions and beneficial effects of the present invention more clear, the present invention provides the following drawings for illustration:

[0055] FIG1 is a structural diagram of a multi-surface aberration compensator scheme 1.

[0056] FIG2 is a schematic diagram of the structure of the second multi-surface aberration compensator scheme.

[0057] Figure 3 is a schematic diagram of the flow channel structure.

[0058] FIG4 is a schematic diagram of the aberration adjustment mirror structure.

[0059] FIG5 is a schematic diagram of the state of the aberration adjustment mirror.

[0060] FIG6 is a schematic diagram of the image quality of a biochip in the prior art.

[0061] FIG7 is a schematic diagram of the radius of the Airy disk at a wavelength of 562 nm.

[0062] FIG8 is a schematic diagram of the GEO radius of the third field of view.

[0063] FIG9 is a schematic diagram of the imaging aberration effect of the adjustment mirror.

[0064] FIG10 is a schematic diagram showing the imaging quality of the first surface.

[0065] FIG11 is a schematic diagram showing the imaging quality of the second surface.

[0066] In the figure, there are a radiation light source 1, an incident light path spectroscope 2, an aberration adjustment mirror 3, an objective lens 4, a biochip 5, a carrier stage 6, a reflected light path spectroscope 7, a first tube mirror 8, a first photosensitive device unit 9, a second tube mirror 10, a second photosensitive device unit 11, a third tube mirror 12, a third photosensitive device unit 13, a fourth tube mirror 14, a fourth photosensitive device unit 15; a container 31, a thin film lens 32, an optical fluid material 33, a first state 34, a second state 35, a third state 36; a first surface 51, a second surface 52, a flow channel 53, an alkaloid component 54; a first spectroscope 71, a second spectroscope 72. DETAILED DESCRIPTION

[0067] The present invention will be further described below with reference to the accompanying drawings and specific embodiments so that those skilled in the art can better understand the present invention and implement it. However, the embodiments are not intended to limit the present invention.

[0068] Example 1

[0069] As shown in FIG1 , the gene sequencing system provided with a multi-surface aberration compensator provided in this embodiment includes a gene sequencer, a multi-surface aberration compensator, and a control module for controlling and processing an aberration adjustment mirror;

[0070] The multi-surface aberration compensator includes at least one aberration adjustment mirror for compensating aberrations in the process of light beam imaging;

[0071] The light beam emitted from the incident light path in the gene sequencer is irradiated onto biological samples located on different surfaces through the objective lens of the gene sequencer. The biological samples are excited by the light beam to generate signal light beams. The signal light beams reflected from different surfaces are focused by corresponding aberration adjustment mirrors and then enter the imaging unit to generate images that meet the requirements.

[0072] The aberration adjustment mirror in this embodiment is arranged outside the multi-surface component, that is, outside the flow channel where the biological sample is placed; in this way, the aberration adjustment mirror will not affect the light beam track inside the flow channel;

[0073] The control module is used to obtain the aberration compensation adjustment value of the aberration generated by different surface spacings and adjust the focal length of the multi-surface aberration compensator according to the aberration compensation adjustment value.

[0074] In this embodiment, the different surfaces provided with biological samples include at least a first surface and a second surface; the aberration adjustment mirror has at least a first control state and a second control state after focusing.

[0075] The signal light beam reflected from the biological sample on the first surface can form a required image in the imaging unit after passing through the aberration adjustment mirror in the first control state;

[0076] The signal light beam reflected from the biological sample on the second surface can form a required image in the imaging unit after passing through the aberration adjustment mirror in the second control state;

[0077] In this embodiment, the first surface is the upper surface of the flow channel in the biochip, and the second surface is the lower surface of the flow channel in the biochip;

[0078] In this embodiment, the radiation beam emitted from the incident light path enters the first surface of the flow channel of the biochip through the objective lens. In a first control state of the aberration adjustment mirror, the signal beam reflected from the first surface of the flow channel passes through the aberration adjustment mirror and enters the imaging unit for imaging. When the focal length of the aberration adjustment mirror is changed, in a second control state of the aberration adjustment mirror, the signal beam reflected from the second surface of the flow channel passes through the aberration adjustment mirror and enters the imaging unit for imaging.

[0079] The incident light path includes a light source and an incident light path reflector; the light beam emitted by the light source enters the flow channel of the biochip through the objective lens;

[0080] In this embodiment, if multiple aberration adjustment mirrors are used, the optical axes of the multiple aberration adjustment mirrors are overlapped and combined on the same straight line to form a lens group, and the focusing function is achieved by adjusting the focal length of the focusing lens group.

[0081] The imaging unit in this embodiment can be an imaging unit in a gene sequencer, or a separate imaging unit independent of the gene sequencer. The imaging unit structure described below includes a reflective light path spectroscope, a first tube lens, a first photosensitive device unit, a second tube lens, and a second photosensitive device unit.

[0082] The reflective light path splitter is used to split the light beam reflected from the flow channel collected by the objective lens into a first light beam and a second light beam. The first light beam enters the first tube lens and converges to be received and imaged by the first photosensitive device unit. The second light beam enters the second tube lens and converges to be received and imaged by the second photosensitive device unit.

[0083] As shown in FIG2 , FIG2 is a schematic diagram of the structure of a second scheme of a multi-surface aberration compensator. In this embodiment, the imaging unit further includes a first beam splitter, a third cylindrical lens, and a third photosensitive device unit; a second beam splitter, a fourth cylindrical lens, and a fourth photosensitive device unit;

[0084] The first light beam enters the first beam splitter and is divided into a third light beam and a fourth light beam. The third light beam enters the first tube lens and converges to the first photosensitive device unit for receiving imaging. The fourth light beam enters the third tube lens and converges to the third photosensitive device unit for receiving imaging. At the same time,

[0085] After entering the second beam splitter, the second beam is divided into a fifth beam and a sixth beam. After entering the second tube lens, the fifth beam converges to the second photosensitive device unit for receiving imaging. After entering the fourth tube lens, the sixth beam converges to the fourth photosensitive device unit for receiving imaging.

[0086] The biochip in this embodiment is placed on a carrier.

[0087] As shown in Figure 3, Figure 3 is a schematic diagram of the flow channel structure. In gene sequencing technology, the biochip is provided with a flow channel, and the flow channel includes a first surface (an inner surface located at the upper part of the flow channel) and a second surface (an inner surface located at the lower part of the flow channel). The alkaloid component to be detected is set in the flow channel.

[0088] As shown in FIG4 and FIG5, the aberration adjustment mirror provided in this embodiment is used to provide an optical element with continuously adjustable focal length; it includes a film lens, a container, and a driver;

[0089] The film lens is disposed on a container, and an optical fluid material is disposed in a closed chamber formed between the film lens and the container. The driver applies pressure to the container to cause the film lens to deform, so as to change the focal length of an imaging module composed of the film lens, the optical fluid material, and the container.

[0090] In the thin film lens of this embodiment, the optical fluid material always fills the closed cavity during the deformation process, ensuring that the imaging module can form a liquid chamber lens, thereby achieving a zoom effect.

[0091] The film lens in the aberration adjustment mirror provided in this embodiment is a deformable lens, generally made of an elastic polymer. The film is sealed on a container filled with a liquid medium. When a driver applies pressure to the container, the film lens bends, thereby changing the focal length. In the first state shown in FIG5 , in a relatively unpressurized state, the lens does not bend. As shown in FIG5 , when a certain pressure is applied, the lens bends, and the focal length changes, forming a second state. When the pressure is varied, the lens bends more and the focal length changes again, forming a third state.

[0092] The driver of this embodiment controls the pressure applied to the container through a control circuit. The adjustment current of the driver is associated with the focal length of the lens. In this embodiment, the focal length of the lens obtained by selecting different optical fluid materials is different.

[0093] The optical fluid material in this embodiment can be selected from a material with a refractive index of about 1.1-1.5 and an Abbe number of 90-110 to enable its use in multi-color applications. A material with a refractive index close to that of water can also be selected. The optical fluid material in this embodiment has a refractive index of 1.3 and an Abbe number of 100, which is a low-dispersion material.

[0094] The driver provided in this embodiment includes a voice coil motor and a controller. The voice coil motor, the controller, the thin film lens, and the container constitute a voice coil liquid lens. The voice coil motor generates a driving force on the container under the action of the controller, so that the focal length of the voice coil liquid lens changes.

[0095] The voice coil motor includes a mover component and a stator component; the mover component and the stator component constitute an electromagnetic drive device;

[0096] The axis of the movable component coincides with that of the stator component, and both the movable component and the stator component are electrically connected to a controller; under the control of the controller, the movable component and the stator component work, and under the action of the electromagnetic field, the movable component and the stator component move, thereby providing a driving force effect of linear motion.

[0097] The stator component provided in this embodiment is arranged on the container, and the mover component is arranged on the film lens. The mover component is used to apply a driving force to the film lens, so that the film lens is deformed;

[0098] The mover component in this embodiment can be any one of a coil or a magnet, and the stator component can be any one of a coil or a magnet. When the mover component is a coil, the stator component is a magnet; when the mover component is a magnet, the stator component is a coil; so that the mover component and the stator component can constitute an electromagnetic drive device that can provide electromagnetic force; the electromagnetic drive device can be of two types, namely, a moving coil type and a moving iron type, which generates different driving forces by changing the coil current, and changes the focal length value of the liquid lens under the action of the driving force.

[0099] The thin film lens in this embodiment adopts a non-uniform thickness elastic film structure. The light-transmitting area of ​​the thin film lens is arranged as a convex-flat structure. The refractive index of the liquid filled in the liquid cavity matches the refractive index of the non-uniform thickness elastic film. Therefore, the focal length of the liquid lens in the initial state is infinite, which is approximately a parallel flat plate, greatly improving the zoom range.

[0100] In this embodiment, the movable component adopts a driving ring arranged on the thin film lens. By squeezing or stretching the driving ring, the thin film lens in the light-transmitting area in the middle of the ring is deformed. The movable component is stuck to the driving ring through the liquid. By controlling the size and direction of the coil current, the liquid lens can be driven to produce deformation in different directions and degrees. The curvature of the thin film lens is changed by the voice coil motor, thereby changing the focal length of the lens.

[0101] The multi-surface aberration compensator in this embodiment uses a cemented lens group; the cemented lens group includes a plurality of lenses having a common optical axis, the distance between the lenses in the cemented lens group is adjustable, and the cemented lens group has at least a first control state and a second control state after focusing;

[0102] The signal light beam reflected from the biological sample on the first surface can form a required image in the imaging unit after passing through the cemented lens group in the first control state;

[0103] The signal light beam reflected from the biological sample on the second surface can form a desired image in the imaging unit after passing through the cemented lens group in the second control state;

[0104] The first surface is an upper surface of the flow channel in the biochip, and the second surface is a lower surface of the flow channel in the biochip;

[0105] The radiation light beam emitted from the incident light path enters the first surface of the flow channel of the biochip through the cemented lens group. Under the first control state of the cemented lens group, the signal light beam reflected back from the first surface of the flow channel passes through the cemented lens group and enters the imaging unit for imaging; when the focal length of the cemented lens group is changed, under the second control state of the cemented lens group, the light beam signal reflected back from the second surface of the flow channel passes through the cemented lens group and enters the imaging unit for imaging.

[0106] The cemented lens group in the cemented lens group includes any one or more combinations of the first cemented lens, the second cemented lens, the third cemented lens, and the fourth cemented lens; the spacing between adjacent lenses is adjustable;

[0107] The light beam emitted from the incident light path is irradiated onto biological samples located on different surfaces through an objective lens;

[0108] The distance between the first cemented lens group and the second cemented lens group is adjustable, thereby achieving aberration compensation between the first surface and the second surface of the flow channel;

[0109] The distances between the first cemented lens, the second cemented lens, and the third cemented lens are adjustable, thereby achieving aberration compensation between the first surface and the second surface of the flow channel;

[0110] The distances between the first cemented lens, the second cemented lens, the third cemented lens, and the fourth cemented lens are adjustable, thereby achieving aberration compensation between the first surface and the second surface of the flow channel;

[0111] The first cemented lens, the second cemented lens, the third cemented lens, and the fourth cemented lens are respectively any one of doublets or triplets.

[0112] The cemented lens group further includes a lens barrel and an adjuster; the cemented lens group is arranged in the lens barrel, and the adjuster is used to adjust the distance between each lens in the cemented lens group.

[0113] Example 2

[0114] This embodiment describes a method for achieving multi-surface imaging through an aberration adjustment mirror based on the above-mentioned multi-surface aberration compensator. This method maintains the optical elements in the original imaging system unchanged, and only provides the aberration adjustment mirror in the original optical path system. The aberration adjustment mirror can be used to achieve imaging of multiple surfaces by the same optical imaging system. The multi-surface imaging method provided in this embodiment and applicable to different flow channels includes the following steps:

[0115] placing a biological sample to be tested in the flow channel of the biochip on the carrier;

[0116] Adjusting the focal length of the aberration adjustment mirror to a first state so that a signal light beam generated by the biological sample disposed on the first surface and excited by the radiation light source passes through the first state and is imaged by the imaging unit;

[0117] Re-adjusting the focal length of the aberration adjustment mirror to a second state so that the signal light beam generated by the biological sample disposed on the second surface and excited by the radiation light source passes through the second state and is imaged by the imaging unit;

[0118] Repeat the above steps so that the signal light beams generated by the biological samples on all surfaces are excited by the radiation light source and then focused by the aberration adjustment mirror and imaged by the imaging unit;

[0119] The gene sequencing results are obtained through a gene sequencer based on the imaging data.

[0120] At the same time, this embodiment also provides an imaging method based on the multi-surface aberration compensator, comprising the following steps:

[0121] acquiring position data of different surfaces on which biological samples are placed;

[0122] Calculate compensation adjustment values ​​for generating aberrations based on position data of different surfaces;

[0123] The aberration adjustment mirror is adjusted according to the compensation adjustment value of the aberration to obtain imaging data of the signal light beam generated by the biological sample on different surfaces excited by the radiation light source in the imaging unit;

[0124] The gene sequencing results are obtained through a gene sequencer based on the imaging data.

[0125] The different surfaces of the biological sample are a first surface and a second surface of a biochip flow channel;

[0126] The position data of the different surfaces include a first distance value and a second distance value;

[0127] The first distance value is the distance between the signal light beam generated by the biological sample excited by the radiation light source on the first surface of the flow channel and the imaging unit when imaging;

[0128] The second distance value is the distance between the signal light beam generated by the biological sample excited by the radiation light source and the imaging unit when imaging;

[0129] According to the two multi-surface aberration compensators provided in this embodiment, the imaging process of each system is as follows:

[0130] The first structure is shown in FIG1 . In the first structure, the light beam in the reflected light path is split to obtain two light beam signals, and the two light beam signals are respectively imaged in the corresponding photosensitive device units. The specific process is as follows:

[0131] (1) First, the light beam generated by the radiation light source passes through the reflective light path spectroscope and is reflected into the objective lens. The objective lens is used to collect the signal light beam generated by the first surface base component in the flow channel of the biochip placed on the carrier being excited by the radiation light source. The signal light beam passes through the aberration adjustment mirror in the first state (first state) and enters the reflective light path spectroscope. The reflective light path spectroscope is used to divide the signal light beam collected by the objective lens into a first light beam and a second light beam. After the first light beam enters the tube lens and converges, it is received by the detection photosensitive device unit for imaging. After the second light beam enters the tube lens and converges, it is received by the detection photosensitive device unit for imaging.

[0132] (2) Secondly, the light beam generated by the radiation light source passes through the incident light path spectrometer and is reflected into the objective lens. The objective lens is used to collect the signal light beam generated by the second surface base component in the flow channel of the biochip placed on the carrier being excited by the radiation light source. After passing through the aberration adjustment mirror in the second state (second state), the signal light beam enters the reflected light path spectrometer. The reflected light path spectrometer is used to divide the signal light beam collected by the objective lens into a first light beam and a second light beam. After the first light beam enters the tube lens and converges, it is received by the detection photosensitive device unit for imaging. After the second light beam enters the tube lens and converges, it is received by the detection photosensitive device unit for imaging.

[0133] (3) Repeat steps (1)-(2).

[0134] The second structure is shown in FIG2 . In the second structure, the light beam in the reflected light path is split to obtain two light beam signals, and the two light beam signals are split again to obtain four light beam signals. Each light beam signal is imaged in the corresponding photosensitive device unit. The specific process is as follows:

[0135] (1) First, the light beam generated by the radiation light source passes through the incident light path spectrometer and is reflected into the objective lens. The objective lens is used to collect the signal light beam generated by the first surface base component in the flow channel of the biochip placed on the carrier being excited by the radiation light source. After passing through the aberration adjustment mirror in the first state (first state), the signal light beam enters the reflected light path spectrometer. The reflected light path spectrometer is used to divide the signal light beam collected by the objective lens into a first light beam and a second light beam. The first light beam enters the first spectrometer. The first spectrometer is used to divide the first light beam of the objective lens into a third and a fourth light beam. After the third light beam enters the tube lens and converges, it is received and imaged by the detection photosensitive device unit. After the fourth light beam enters the first tube lens and converges, it is received and imaged by the detection photosensitive device unit. At the same time, the second light beam enters the second spectrometer. The second spectrometer is used to divide the second light beam of the objective lens into a fifth and a sixth light beam. After the fifth light beam enters the tube lens and converges, it is received and imaged by the detection photosensitive device unit. After the sixth light beam enters the second tube lens and converges, it is received and imaged by the detection photosensitive device unit.

[0136] (2) Secondly, the light beam generated by the radiation light source passes through the incident light path spectrometer and is reflected into the objective lens. The objective lens is used to collect the signal light beam generated by the second surface base component in the flow channel of the biochip placed on the carrier being excited by the radiation light source. After passing through the aberration adjustment mirror in the second state (second state), the signal light beam enters the reflected light path spectrometer. The reflected light path spectrometer is used to divide the signal light beam collected by the objective lens into a first light beam and a second light beam. The first light beam enters the first spectrometer. The first spectrometer is used to divide the first light beam of the objective lens into a third and a fourth light beam. After the third light beam enters the tube lens and converges, it is received and imaged by the detection photosensitive device unit. After the fourth light beam enters the first tube lens and converges, it is received and imaged by the detection photosensitive device unit. At the same time, the second light beam enters the second spectrometer. The second spectrometer is used to divide the second light beam of the objective lens into a fifth and a sixth light beam. After the fifth light beam enters the tube lens and converges, it is received and imaged by the detection photosensitive device unit. After the sixth light beam enters the second tube lens and converges, it is received and imaged by the detection photosensitive device unit.

[0137] (3) Repeat steps (1)-(2).

[0138] The aberration adjustment mirror in this embodiment is used to achieve continuous adjustment of the aberration; thereby achieving imaging compensation on the double-layer surface or multi-layer surface of the biochip flow channel, or allowing the upper / lower surfaces of double-layer chips with different flow channel thicknesses to be imaged simultaneously.

[0139] Example 3

[0140] This embodiment describes in detail the comparative analysis of gene sequencing using a variable-focus multi-surface aberration compensator in conjunction with specific implementation methods.

[0141] As shown in FIG6 , it is a schematic diagram of an image quality image of the first surface and the second surface of a biochip in the prior art. In the example, the microscopic imaging device images a biochip 5 with a cover glass thickness of 300 μm and a flow channel thickness of 100 μm, wherein the objective lens parameters are as follows: NA is 0.8, field of view is 1.2 mm, and magnification is 20 times.

[0142] As shown in Figure 7, the radius of the Airy disk at a wavelength of 562nm (i.e., the optical diffraction limit) is 9.166um. If the GEO spot radius of each field of view obtained from the spot diagram is greater than the diffraction limit, then for an infinitesimal point, its spot radius after passing through the optical system is the GEO radius. If it is less than the diffraction limit, then the spot radius is the diffraction limit. For example, in the imaging quality of the first surface, the GEO radius of the third field of view is 4.032um, which is smaller than the Airy disk radius. Therefore, its spot radius is the Airy disk radius, i.e., 9.166um. In the imaging quality of the second surface,

[0143] As shown in Figure 8, the GEO radius of the third field of view is 71.387 μm, which is much larger than the Airy disk radius. Therefore, its spot radius is equal to its GEO radius of 71.387 μm. It should be understood that the aforementioned light wavelength, cover glass thickness, flow channel thickness, objective lens parameters, and Airy disk radius are merely examples and do not limit these parameters in this embodiment. These parameters can be set according to actual needs.

[0144] As shown in FIG9 , the aberration-adjusting mirror of this embodiment is a liquid lens, the curvature of which can be changed by a voice coil motor control, thereby changing the focal length of the lens, and the lens material is a low-dispersion material with a refractive index of 1.3 and an Abbe number of 100. When imaging the first surface signal, the curvature of the liquid lens is infinite and is equivalent to a flat liquid material. When imaging the second surface signal, the focal length is changed to compensate for the aberration. Moreover, since there is a flow channel solution between the first and second surfaces, the imaging will be subjected to large aberrations, mainly spherical aberration, so that the imaging quality is reduced. The aberration can be well compensated by changing the curvature of the liquid lens.

[0145] Figure 10 shows a schematic diagram of image quality for imaging the first and second surfaces of a biochip in the prior art. In this example, a microscopic imaging device images a biochip with a 300 μm cover glass thickness and a 100 μm flow channel thickness. The objective lens parameters are as follows: NA 0.8, field of view 1.2 mm, and magnification 22.22x. The figure shows that the Airy disk radius (i.e., the optical diffraction limit) at a wavelength of 562 nm is 9.53 μm. If the GEO spot radius of each field of view obtained from the point diagram is greater than the diffraction limit, then for an infinitesimal point, the spot radius after passing through the optical system is the GEO radius. If it is less than the diffraction limit, then the spot radius is the diffraction limit. For example, in the image quality of the first surface, the GEO radius of the third field of view is 3.526 μm, which is smaller than the Airy disk radius. Therefore, its spot radius is the Airy disk radius, i.e., 9.53 μm. In the image quality of the second surface, the GEO radius of the third field of view is 3.526 μm, which is smaller than the Airy disk radius.

[0146] As shown in Figure 11, the GEO radius of the third field of view is 6.241 μm, much larger than the radius of the Airy disk. Therefore, its spot radius is equal to its GEO radius of 9.53 μm. After adding the aberration adjustment mirror 3, the image quality remains unchanged, indicating that the compensation group produces a good aberration compensation effect.

[0147] The aberration adjustment mirror selected in this embodiment has an adjustment range of 0-150 μm, which can achieve continuous adjustment of the aberration within the range of 0-150 μm. When the flow channel thickness of the biochip is in the range of 0-120 μm, plus the spherical aberration generated by the liquid medium in the flow channel is about 30 μm, the thickness of the biochip can fluctuate from 0-150 μm; thus, this technical solution can be applied to chips produced by different manufacturers (chips with different flow channel thicknesses). Directly adjusting the adjustment range of the aberration adjustment mirror can achieve imaging of the first and second surfaces of the flow channels of different chips without replacing the optical components in the imaging system, thus having a wide range of applicability.

[0148] The incident light path spectrometer in this embodiment can have a reflectance-to-transmission ratio of 1:1, or other splitting ratios; a bandpass filter (not shown) can be placed in front of the barrel lens or behind the barrel lens to select the wavelength of light passing through the barrel lens / before the imaging sensor; the material used for the aberration adjustment mirror can be a liquid-filled liquid lens or a double-liquid electrowetting liquid lens.

[0149] The above embodiments are merely preferred embodiments for the purpose of fully illustrating the present invention, and the scope of protection of the present invention is not limited thereto. Equivalent substitutions or modifications made by those skilled in the art based on the present invention are within the scope of protection of the present invention. The scope of protection of the present invention shall be subject to the claims.

Claims

1. A gene sequencing system provided with a multi-surface aberration compensator, including a gene sequencer, characterized in that: Also included is a multi-surface aberration compensator; The multi-surface aberration compensator includes at least one aberration adjustment mirror for compensating for aberrations generated by a signal beam of a gene sequencer during an imaging process; The light beam emitted from the incident light path in the gene sequencer is irradiated to biological samples located on different surfaces through the objective lens of the gene sequencer. The biological samples are excited by the light beam to generate signal light beams. The signal light beams reflected back from different surfaces are focused by corresponding aberration adjustment mirrors and then enter the imaging unit to generate images.

2. The gene sequencing system provided with a multi-surface aberration compensator according to claim 1, characterized in that: It also includes a control module for controlling the aberration adjustment mirror, wherein the control module is used to obtain the aberration compensation adjustment value of the aberration generated by different surface spacings and adjust the focal length of the multi-surface aberration compensator according to the aberration compensation adjustment value.

3. The gene sequencing system provided with a multi-surface aberration compensator according to claim 1, characterized in that: The different surfaces are the first surface and the second surface of the biochip flow channel arranged on the upper gene sequencer; the first surface is the upper surface of the flow channel in the biochip, and the second surface is the lower surface of the flow channel in the biochip.

4. The gene sequencing system provided with a multi-surface aberration compensator as claimed in claim 3, characterized in that: The radiation light beam emitted by the incident light path enters the first surface of the flow channel of the biochip through the objective lens. In the first control state of the aberration adjustment mirror, the signal light beam reflected back from the first surface of the flow channel passes through the aberration adjustment mirror and enters the imaging unit for imaging; when the focal length of the aberration adjustment mirror is changed, in the second control state of the aberration adjustment mirror, the light beam signal reflected back from the second surface of the flow channel passes through the aberration adjustment mirror and enters the imaging unit for imaging.

5. The gene sequencing system provided with a multi-surface aberration compensator according to claim 1, characterized in that: The imaging unit comprises a reflective light path spectroscope, a first tube lens, a first photosensitive device unit, a second tube lens, and a second photosensitive device unit; The reflective light path splitter is used to split the light beam reflected from the flow channel collected by the objective lens into a first light beam and a second light beam. The first light beam enters the first tube lens and converges to be received and imaged by the first photosensitive device unit. The second light beam enters the second tube lens and converges to be received and imaged by the second photosensitive device unit.

6. The gene sequencing system provided with a multi-surface aberration compensator according to claim 5, characterized in that: The imaging unit further includes a first beam splitter, a third tube lens, and a third photosensitive device unit; a second beam splitter, a fourth tube lens, and a fourth photosensitive device unit; The first light beam enters the first beam splitter and is divided into a third light beam and a fourth light beam. The third light beam enters the first tube lens and converges to the first photosensitive device unit for receiving imaging. The fourth light beam enters the third tube lens and converges to the third photosensitive device unit for receiving imaging. At the same time, The second light beam enters the second beam splitter and is divided into a fifth light beam and a sixth light beam. The fifth light beam enters the second tube lens and converges The sixth light beam enters the fourth tube lens and converges to the fourth photosensitive device unit for receiving imaging.

7. The gene sequencing system provided with a multi-surface aberration compensator according to claim 1, characterized in that: The aberration adjustment mirror comprises a film lens, a container and a driver; The film lens is arranged on a container, and an optical fluid material is arranged in a closed chamber formed between the film lens and the container. The driver applies pressure to the container and causes the film lens to deform so as to change the focal length of an imaging module composed of the film lens, the optical fluid material and the container. The film lens in the aberration adjustment mirror is a deformable lens, and the film lens is made of an elastic polymer. The film lens is sealed on a container filled with a liquid medium, and the driver applies pressure to the container to cause the film lens to bend so that the focal length changes.

8. The gene sequencing system provided with a multi-surface aberration compensator according to claim 7, characterized in that: The optical fluid material is selected from materials with a refractive index of about 1.1-1.5 and an Abbe number of 90-110.

9. The gene sequencing system provided with a multi-surface aberration compensator according to claim 7, characterized in that: The driver includes a voice coil motor and a controller. The voice coil motor, the controller, the thin film lens, and the container constitute a voice coil liquid lens. The voice coil motor generates a driving force on the container under the action of the controller, so that the focal length of the voice coil liquid lens changes.

10. The gene sequencing system provided with a multi-surface aberration compensator according to claim 9, characterized in that: The voice coil motor comprises a mover component and a stator component; the mover component and the stator component constitute an electromagnetic drive device; The axis of the movable component coincides with that of the stator component, and both the movable component and the stator component are electrically connected to a controller; under the control of the controller, the movable component and the stator component work, and under the action of the electromagnetic field, the movable component and the stator component move, thereby providing a driving force for linear motion; The stator component is arranged on the container, the mover component is arranged on the film lens, and the mover component is used to apply a driving force to the film lens so that the film lens is deformed; The movable component adopts any one of a coil or a magnet, and the stator component adopts any one of a coil or a magnet. When the movable component adopts a coil, the stator component is a magnet; when the movable component adopts a magnet, the stator component is a coil; so that the movable component and the stator component can constitute an electromagnetic drive device that can provide electromagnetic force.

11. The gene sequencing system provided with a multi-surface aberration compensator according to claim 10, characterized in that: The electromagnetic driving device adopts two types: moving coil type and moving iron type. Different driving forces are generated by changing the coil current, and the focal length value of the liquid lens is changed under the action of the driving force.

12. The gene sequencing system provided with a multi-surface aberration compensator according to claim 7, characterized in that: The film lens adopts a non-uniform thickness elastic film structure, the light-transmitting area of ​​the film lens is arranged as a convex-flat structure, and the refractive index of the liquid filled in the liquid cavity matches the refractive index of the non-uniform thickness elastic film.

13. The gene sequencing system provided with a multi-surface aberration compensator according to claim 10, characterized in that: The movable member adopts a driving ring arranged on the film lens, and drives the film lens in the light-transmitting area in the middle of the ring to deform by squeezing or stretching the driving ring; The movable component is adhered to the driving ring through liquid. By controlling the size and direction of the coil current, the liquid lens can be driven to produce deformations in different directions and degrees. The curvature of the thin film lens is changed by the voice coil motor control, thereby changing the focal length of the lens.

14. The gene sequencing system provided with a multi-surface aberration compensator according to claim 1, characterized in that: The multi-surface aberration compensator adopts a cemented lens group; the cemented lens group includes a plurality of lenses with a common optical axis, and the distance between the lenses in the cemented lens group is adjustable.

15. The gene sequencing system provided with a multi-surface aberration compensator according to claim 14, characterized in that: The cemented lens group has at least a first control state and a second control state after focusing; The signal light beam reflected from the biological sample on the first surface can form an image in the imaging unit after passing through the cemented lens group in the first control state; The signal light beam reflected from the biological sample on the second surface passes through the cemented lens group in the second control state and then enters the imaging unit to generate an image; The first surface is an upper surface of a flow channel in the biochip, and the second surface is a lower surface of a flow channel in the biochip; The radiation light beam emitted from the incident light path enters the first surface of the flow channel of the biochip through the glued lens group. Under the first control state of the glued lens group, the signal light beam reflected from the first surface of the flow channel passes through the glued lens group and enters the imaging unit for imaging; when the focal length of the glued lens group is changed, under the second control state of the glued lens group, the light beam signal reflected from the second surface of the flow channel passes through the glued lens group and enters the imaging unit for imaging.

16. The gene sequencing system provided with a multi-surface aberration compensator according to claim 14, characterized in that: The cemented lens group in the cemented lens group includes any one or more combinations of the first cemented lens, the second cemented lens, the third cemented lens, and the fourth cemented lens; the first cemented lens, the second cemented lens, the third cemented lens, and the fourth cemented lens are any one of double cemented lenses or triple cemented lenses.

17. The gene sequencing system provided with a multi-surface aberration compensator according to claim 14, characterized in that: The cemented lens group also includes a lens barrel and an adjuster; the cemented lens group is arranged in the lens barrel, and the adjuster is used to adjust the distance between each lens in the cemented lens group.

18. A method for implementing gene sequencing using the gene sequencing system according to any one of claims 1 to 17, characterized in that: The following steps are involved: Placing the biological sample to be tested in the flow channel of the biochip on the stage; The focal length of the aberration adjustment mirror is adjusted to a first state, so that a signal light beam generated by the biological sample disposed on the first surface and excited by the radiation light source passes through the first state and is imaged in the imaging unit; Re-adjusting the focal length of the aberration adjustment mirror to a second state, so that the signal light beam generated by the biological sample disposed on the second surface and excited by the radiation light source passes through the second state and is imaged in the imaging unit; Repeat the above steps so that the signal light beams generated by the biological samples on all surfaces being excited by the radiation light source are focused by the aberration adjustment mirror and then imaged in the imaging unit; The gene sequencing results are obtained through a gene sequencer based on the imaging data.

19. The gene sequencing method with a multi-surface aberration compensator as claimed in claim 18, characterized in that: The signal light beam passes through a beam splitter to obtain two light beam signals, and the two light beam signals are respectively imaged in corresponding photosensitive device units.

20. The gene sequencing method with a multi-surface aberration compensator as claimed in claim 19, characterized in that: The signal light beam is passed through a spectroscope to obtain two light beam signals, which are then split again through a spectroscope to obtain four light beam signals, and each light beam signal is imaged in a corresponding photosensitive device unit.

21. A method for implementing gene sequencing using the gene sequencing system according to any one of claims 1 to 17, characterized in that: The following steps are involved: acquiring position data of different surfaces on which the biological sample is disposed; Calculate compensation adjustment values ​​for generating aberrations according to position data of different surfaces; The aberration adjustment mirror is adjusted according to the compensation adjustment value of the aberration to respectively obtain imaging data of the signal light beam generated by the biological samples on different surfaces excited by the radiation light source in the imaging unit; The gene sequencing results are obtained through a gene sequencer based on the imaging data.

22. The gene sequencing method with a multi-surface aberration compensator according to claim 21, characterized in that: The different surfaces of the biological sample are a first surface and a second surface of a biochip flow channel; The position data of the different surfaces include a first distance value and a second distance value; The first distance value is the distance of the signal light beam generated by the biological sample excited by the radiation light source on the first surface of the flow channel when the imaging unit images the signal light beam; The second distance value is the distance of the signal light beam generated by the biological sample excited by the radiation light source on the second surface of the flow channel when the imaging unit forms an image.

Images