Optical system for a gene sequencer and image correction device

Abstract

The application provides an optical system for a gene sequencer and an image correction device. The optical system comprises a sample module, an illumination module, a focusing module, a light splitting module and an imaging module. The image correction device is used in the imaging module of the optical system and comprises a connecting plate, an image sensor adjusting device and a reflection adjusting mirror. The connecting plate is connected with an image sensor in the imaging module of the optical system. The image sensor is detachably mounted on the connecting plate. The connecting plate is connected with the image sensor adjusting device. The reflection adjusting mirror is arranged in front of the image sensor. The reflection adjusting mirror can be adjusted to rotate around an optical axis and perpendicular to the optical axis to correct the image. The system and the device in the application are simple and reliable in adjustment operation, can effectively correct the image, and can more satisfy the imaging accuracy through the multi-degree-of-freedom image adjustment and correction. The application plays an important role in analyzing base types and base sequences.

Description

TECHNICAL FIELD

[0001] The present application mainly relates to the field of optical system design, image correction and gene sequencing technology, and particularly relates to an optical system for a gene sequencer and an image correction device. BACKGROUND

[0002] At present, the design research on the optical system of the gene sequencer mainly includes the design of the illumination system, the selection of the light splitting mirror group and the design of the imaging device. When the existing direct illumination system irradiates the sample, the illumination spot will be uneven, thereby causing the imaging picture quality to decrease and the sequencing accuracy to decrease, so it is meaningful to reasonably consider the illumination system for the gene sequencing process. Improper selection of the light splitting mirror group will directly cause the weakening or even loss of the sequencing information, according to the fluorescence information emitted by the excited sample, reasonable selection of the light splitting mirror group can effectively collect the sequencing information to achieve the sequencing purpose. In addition, reasonable consideration of the spatial arrangement of the optical elements can minimize the occupied space of the optical system, and the smaller the occupied space of the optical system, the more compact the entire product.

[0003] In the base sequencing judgment process, the pixel consistency of the photographed image has a high requirement, and the pixel consistency refers to the coordinate position of the photographed image as much as possible to coincide. In order to identify the type of the base to be sequenced, a large number of base images need to be obtained, and the base images are obtained by a professional CCD camera. Due to the positioning error of each element of the system, the self-positioning error of the camera and the fact that the image field of the different base types photographed by the cameras does not meet the pixel consistency during the photographing process of the CCD camera, which will bring the consequence that the correct base sequence cannot be analyzed in the subsequent image processing and base recognition operations, resulting in sequencing failure. The existing technology uses an algorithm to compensate, for example, the camera calibration method and the image registration method mentioned in patent CN201810032793.3, which can perform image registration, but when the relative positions of the images photographed by different CCD cameras are too different, this method is not applicable, so it is necessary to design and propose an image correction device, which plays an important role in subsequent analysis of base types and base sequences. SUMMARY

[0004] The embodiment of the present application provides an optical system for a gene sequencer and an image correction device, and the optical system comprises:

[0005] The sample module comprises a sample;

[0006] The illumination module comprises a light source, a lens group, a laser dichroic mirror, a laser color filter and an objective lens;

[0007] The focusing module comprises a focusing module, a focusing dichroic mirror and a focusing color filter;

[0008] The light splitting module comprises a light splitting dichroic mirror set;

[0009] The imaging module comprises a tube lens, an image sensor, an image correction device, and a color filter;

[0010] The illumination module comprises a light source, a lens set, a laser dichroic mirror, a laser color filter, and an objective lens; the light source is used to emit excitation light for exciting a sample; the lens set comprises a collimating mirror, an imaging lens, a laser dichroic mirror, and an objective lens; the collimating mirror is arranged in the optical path between the light source and the laser dichroic mirror, and the collimating mirror is used to expand the beam of the excitation light from the light source; the imaging lens is arranged in the optical path between the light source and the laser dichroic mirror, and the imaging lens is used to image the expanded excitation light on the back focal plane of the objective lens; the laser dichroic mirror is arranged in the optical path between the objective lens and the tube lens, and the laser dichroic mirror is used to reflect the excitation light from the light source into the objective lens to excite the sample and transmit the emission light from the sample into the image sensor; the laser color filter is arranged in the optical path between the light source and the laser dichroic mirror, and the laser color filter is used to ensure the cleanliness of the excitation light from the light source; and the objective lens collects the excitation light for exciting the sample to the sample and collects the emission light from the sample to the imaging module.

[0011] Further, the focusing module emits focusing laser light, the focusing color filter is arranged in the optical path between the focusing module and the focusing dichroic mirror, the focusing color filter is used to ensure the cleanliness of the focusing laser light, the focusing dichroic mirror is arranged in the optical path between the objective lens and the tube lens, the focusing dichroic mirror reflects the focusing laser light into the objective lens to the sample, adjusts the vertical relative position of the objective lens and the sample, the sample reflects the focusing laser light back to the focusing module to determine the vertical relative position of the objective lens and the sample, so as to achieve the purpose of focusing.

[0012] Further, the light splitting module comprises a dichroic mirror set, the dichroic mirror set comprises a first dichroic mirror, a second dichroic mirror, a third dichroic mirror, and a color filter for blocking specific emission light from the sample; the dichroic mirror set is arranged in the optical path between the objective lens and the tube lens, and the dichroic mirror is used to transmit one of the excitation light and the emission light and reflect the other of the excitation light and the emission light.

[0013] Further, the above optical system is photographed by the image sensor in the imaging module, and the difference of the image pixels photographed by the image sensor is adjusted based on the difference of the characteristic bright spots of the photographed image.

[0014] Further, the image correction device is further included in the imaging module of the optical system, the image correction device comprises a connecting plate, an image sensor adjusting device and a reflection adjusting mirror, the connecting plate is connected with the image sensor in the imaging module of the optical system, the image sensor is detachably mounted on the connecting plate, the connecting plate is connected with the image sensor adjusting device, the image sensor adjusting device comprises a first adjusting vertical plate, a first adjusting screw, a second adjusting bottom plate, a second adjusting screw, a third adjusting bottom plate and a third adjusting screw, the first adjusting vertical plate is used for adjusting the first direction freedom degree, the second adjusting bottom plate is used for adjusting the second direction freedom degree, the third adjusting bottom plate is used for adjusting the third direction freedom degree, the reflection adjusting mirror is arranged in front of the image sensor, and the reflection adjusting mirror adjusts the rotation direction around the optical axis and the rotation direction perpendicular to the optical axis to correct the image.

[0015] The system and the device in the application are simple and reliable in adjustment operation, can effectively correct the image, and can more meet the imaging precision through the multi-freedom degree image adjustment and correction, thereby playing an important role in analyzing the base type and the base sequence. BRIEF DESCRIPTION OF DRAWINGS

[0016] Figure 1 is a schematic diagram of the optical system of the application;

[0017] Figure 2 is a schematic diagram of the module composition of the optical system of the application;

[0018] Figure 3 is an image and a fluorescent feature point photographed by the image sensor in the optical system of the application;

[0019] Figure 4 is a schematic diagram of the image correction device model in the optical system of the application;

[0020] Figure 5 is a schematic diagram of the image correction device model in the optical system of the application;

[0021] Figure 6 is a schematic diagram of the connecting plate model in the image correction device in the optical system of the application;

[0022] Figure 7 is a schematic diagram of the first adjusting vertical plate model in the image correction device in the optical system of the application;

[0023] Figure 8 is a schematic diagram of the second adjusting bottom plate model in the image correction device in the optical system of the application;

[0024] Figure 9 is a schematic diagram of the third adjusting bottom plate model in the image correction device in the optical system of the application;

[0025] Figure 10 This is a schematic diagram of the positioning key and fixing key of the image correction device in the optical system of the present invention;

[0026] In the diagram: 1-Sample, 2-Objective lens, 3-Laser dichroic mirror, 4-First dichroic mirror, 5-Laser imaging lens, 6-Collimating lens, 7-Laser, 8-Second dichroic mirror, 9-First tube lens, 10-First reflection adjustment mirror, 11-First color filter, 12-First camera, 13-Second tube lens, 14-Fourth camera, 15-Fourth color filter, 16-Second reflection adjustment mirror, 17-Fourth tube lens, 18-Third dichroic mirror, 19-Third tube lens, 20- Third reflecting adjustment mirror, 21-Second color filter, 22-Second camera, 23-Third color filter, 24-Third camera, 25-Focusing dichroic mirror, 26-Focusing color filter, 27-Focusing module, 28-Laser color filter, 101-First adjusting screw, 103-Second adjusting screw, Screw 102-Third adjustment, 200-Third base plate, 201-Second base plate, 202-Camera connecting plate, 203-First vertical plate, 204-Positioning key, 204-1-Fixing slot.

[0027] 205 - First fixing key, 206 - Second fixing key, 1000 - Fluorescent spot, 2000 - Background, 3000 - Fluorescent feature point, 1b - First threaded hole, 2b - Second threaded hole, 4b - Third threaded hole, 5b - Fourth threaded hole, 7b - Fifth threaded hole, 8b - Sixth threaded hole, 101a - Seventh threaded hole, 102a - Eighth threaded hole, 103a - Ninth threaded hole, 102b - First nut groove, 103b - Second nut groove, 101b - Third nut groove 1a-First sliding groove, 4a-Second sliding groove, 204-2-Third sliding groove, 2a-First through hole, 3a-Second through hole, 5a-Third through hole, 6a-Fourth through hole, 7a-Fifth through hole, 111-First screw, 222-Second screw, 333-Third screw, 444-Fourth screw, 555-Fifth screw, 666-Sixth screw, 777-Seventh screw, I-Sample module, II-Illumination module, III-Focusing module, IV-Spectrometer module, V-Imaging module. Detailed Implementation

[0028] The preferred embodiments of the present invention will now be described in detail with reference to the accompanying drawings, so that the advantages and features of the present invention can be more easily understood by those skilled in the art, thereby providing a clearer and more explicit definition of the scope of protection of the present invention.

[0029] In the following description, a lot of specific details are set forth in order to provide a thorough understanding of the embodiments of the present application, and it will be apparent to those skilled in the art that the application can be practiced without some or all of these specific details. In other instances, well known process steps have not been described in detail in order not to unnecessarily obscure the present application. The specific embodiments of the present application are described in order to provide a thorough and enabling disclosure of the present application.

[0030] The present application provides an optical system for a gene sequencer and an image correction device, the optical system schematic diagram is as Figure 1 , the optical system module composition schematic diagram is as Figure 2 .

[0031] The optical system is divided into five modules: sample module I, focusing module II, illumination module III, light splitting module IV and imaging module V receiving image information.

[0032] As Figure 1 the optical system schematic diagram, according to the module composition schematic diagram Figure 2 classification, sample module I includes sample 1, sample 1 usually carries four fluorescent markers, such as A dye molecules, B dye molecules, C dye molecules and D dye molecules, the four fluorescent excitation wavelengths are λ1, λ1, λ2 and λ2, and the wave peaks of the wave bands of the fluorescent emission wavelengths are λ3, λ4, λ5 and λ6; illumination module III includes laser 7, which emits excitation light beams of λ1 and λ2 required for fluorescent excitation, the excitation light beams are projected through laser color filter 28, expanded through collimating mirror 6, and imaged at the back focal plane of objective lens 2 of optical fiber imaging, in the imaging process, the light beams are reflected by laser dichroic mirror 3 and transmitted through focusing dichroic mirror 25 to enter objective lens 2, and the two excitation light beams of different wavelengths are uniformly irradiated on sample 1 through objective lens 2.

[0033] Sample 1 is excited by the excitation light beams to emit different fluorescent light, the different fluorescent light is divided into several paths through light splitting module IV, the first reflecting adjusting mirror 10, the second reflecting adjusting mirror 16 and the third reflecting adjusting mirror 20 in the imaging module of the optical system are in 45-degree turning state, according to the comparison of the images, the first reflecting adjusting mirror 10, the second reflecting adjusting mirror 16, the third reflecting adjusting mirror 20 and the image correction device correct the pixels of the images, and the imaging module V is used for imaging and photographing, so that the pixel difference of the four camera image fluorescent feature points 3000 is within 15 pixels, and we consider that the image field of view photographed by the camera is coincident.

[0034] Before taking pictures, focusing module 27 in focusing module II emits focusing laser, which is reflected by focusing dichroic mirror 25 after passing through focusing color filter 26, received by objective lens, and then enters sample and is reflected back to objective lens, the vertical distance between objective lens and sample is adjusted until the focusing module judges that the sample is on the working surface of objective lens, and then the focusing is considered to be completed.

[0035] After focusing, the optical system takes a picture of the fluorescence as shown Figure 3 The fluorescence bright spot 1000 is the genetic base information, the black area is the background 2000, and the white circle part is the fluorescence feature point 3000. According to the images taken by each camera, the pixel position of the fluorescence feature point 3000 in the taken image is determined by using software (such as imagej image processing software), and then the pixel distance that each camera needs to adjust is obtained. The image correction device is adjusted to achieve the purpose of image pixel consistency.

[0036] The functions of the first dichroic mirror 4, the second dichroic mirror 8, and the third dichroic mirror 18 in the optical system schematic diagram are to reflect the sample fluorescence emission light, such as the above-mentioned waveband with a wave peak of λ3 and λ4 fluorescence, and to transmit the sample fluorescence emission light, such as the above-mentioned waveband with a wave peak of λ5 and λ6 fluorescence. The first camera 12, the second camera 22, the third camera 24, and the fourth camera 14 in the imaging module V of the optical system schematic diagram are connected with the camera connecting plate 202 in the image correction device by screws, which are used to adjust the camera pixel consistency.

[0037] The imaging module V includes the third color filter 23 and the third camera 24, the fourth color filter 15 and the fourth camera 14; the imaging module V includes the second color filter 21 and the second camera 22, the first color filter 11 and the first camera 12. The light splitting module IV includes the first dichroic mirror 4, the second dichroic mirror 8 and the third dichroic mirror 18, the first dichroic mirror 4 reflects the two kinds of fluorescence excited by the laser 7 with the wavelength λ1 into the imaging module V, and transmits the two kinds of fluorescence excited by the laser 7 with the wavelength λ2 into the imaging module V; the second dichroic mirror 8 reflects one kind of fluorescence excited by λ1, and the image is formed through the second tube lens 13, and before the camera, the second color filter 21 is passed through to block the stray light except the fluorescence into the second camera 22; the second dichroic mirror 8 transmits one kind of fluorescence excited by λ1, and the image is formed through the first tube lens 9, and the first reflection adjusting mirror 10 in the image correction device in the imaging module V preliminarily corrects the image pixel consistency, and can shorten the optical system occupied space, so that the product can be more compact, and the pixel consistency is fine adjusted through the image correction device, and the first color filter 11 is passed through to block the stray light except the fluorescence into the first camera 12. The third dichroic mirror 18 reflects one kind of fluorescence excited by λ2, and the image is formed through the fourth tube lens 17, and the second reflection adjusting mirror 16 in the image correction device in the imaging module V preliminarily corrects the image pixel consistency, and the pixel consistency is fine adjusted through the image correction device, and the fourth color filter 15 is passed through to block the stray light except the fluorescence into the fourth camera 14; the third dichroic mirror 18 transmits one kind of fluorescence excited by λ2, and the image is formed through the third tube lens 19, and the third reflection adjusting mirror 20 in the image correction device in the imaging module V preliminarily corrects the image pixel consistency, and the pixel consistency is fine adjusted through the image correction device, and the third color filter 23 is passed through before the camera to block the stray light except the fluorescence into the third camera 24. The structure model diagram of the image correction device is as shown in Figure 4 , and the structure model detail diagram of the image correction device is as shown in Figure 5 .

[0038] The structure part of the image correction device is as shown in

[0039] The image correction device in the embodiment includes a connecting plate, an image sensor adjusting device and a reflection adjusting mirror. The connecting plate is connected with the image sensor in the optical system imaging module, the reflection adjusting mirror is arranged in front of the image sensor, and the reflection adjusting mirror adjusts the rotation direction around the optical axis and the rotation direction perpendicular to the optical axis to correct the image. Taking the coordinate system direction at the lower right corner of the figure as an example, the image sensor adjusting device mainly includes a camera connecting plate 202 (the model diagram is as shown in Figure 6 ), a first vertical plate 203 (the model diagram is as shown in Figure 7), adjusting the second direction freedom (Y direction) of the second bottom plate 201 (model schematic diagram as shown in Figure 8 ), adjusting the third direction freedom (X direction) of the third bottom plate 200 (model schematic diagram as shown in Figure 9 ), and positioning the key 204, the first fixed key 205 and the second fixed key 206 (model schematic diagram as shown in Figure 10 ).

[0040] The image sensor adjusting device can realize multi-freedom adjustment of X, Z and Y in front and back, up and down, and left and right, and is simple to operate while meeting the regulation and control requirements. The detailed assembly diagram of the device is shown in the figure. The third bottom plate 200 is provided with four first threaded holes 1b, and the second bottom plate 201 is provided with a first sliding groove 1a, and the length of the first sliding groove 1a is the range of the Y freedom. The first screw 111 passes through the first sliding groove 1a and cooperates with the first threaded hole 1b to play a role of fixing the position, the third bottom plate 200 is provided with a fixed groove 204-1 at the bottom, the positioning key 204 is fixed with the whole machine panel, and the third bottom plate 200 cooperates with the positioning key 204 through the fixed groove 204-1 to be positioned, and the purpose is that when the position of the third bottom plate 200 is adjusted, the third bottom plate 200 can move straight to the adjusting direction, avoiding the rotation of the third bottom plate 200 to cause adjustment error; the second bottom plate 201 is provided with two second threaded holes 1b, the first vertical plate 203 is provided with a first through hole 2a, and the second screw 222 passes through the first through hole 2a of the first vertical plate 203 and cooperates with the second threaded hole 1b of the second bottom plate 201 to be fixed; the first vertical plate 203 is provided with a second sliding groove 4a, and the length of the second sliding groove 4a is the range of the Z freedom, the camera connecting plate 202 is provided with a third threaded hole 4b, and the fourth screw 444 passes through the second sliding groove 4a of the first vertical plate 203 and cooperates with the third threaded hole 4b of the camera connecting plate 202 to be fixed; the third screw 333 passes through the second through hole 3a of the camera connecting plate 202 and cooperates with the threaded hole of the camera to be fixed. The positioning key 204 is fixed on the whole machine panel (here the whole machine panel is not drawn) and cooperates with the sliding groove 204-1 in the third bottom plate 200; the other two positioning keys 204 pass through the third through hole 5a and 7a and cooperate with the fourth threaded hole 5b and the fifth threaded hole 7b on the third bottom plate 200 through the fifth screw 555 and the seventh screw 777 to be fixed, one of the positioning keys 204 cooperates with the third sliding groove 204-2 on the second bottom plate 201, and the positioning key 204 cooperates with the third adjusting screw 103. The second fixed key 206 is provided with a fourth through hole 6a, and the screw 888 passes through the fourth through hole 6a of the fixed part 206 and cooperates with the sixth threaded hole 8b on the third bottom plate 200 to be fixed; the first fixed key 205 is provided with a fourth through hole 6a, and the sixth screw 666 passes through the fourth through hole 6a of the fixed part 205 and cooperates with the threaded hole on the whole machine panel to be fixed.

[0041] The first adjusting screw 101 is fixed with the seventh threaded hole 101a of the first vertical plate 203, the nut of the adjusting screw is clamped in the third nut slot 101b on the camera connecting plate 202, when the first direction freedom (Z direction) needs to be adjusted, the first adjusting screw 101 and the second threaded hole 101a of the first vertical plate 203 are used to adjust the length, the nut of the first adjusting screw 101 moves the camera connecting plate together with the camera to the Z direction to achieve the adjusting purpose.

[0042] The second adjusting screw 103 is fixed with the ninth threaded hole 103a of the fixed key 204, the nut of the adjusting screw is clamped in the second nut slot 103b on the second bottom plate 201, when the second direction freedom (Y direction) needs to be adjusted, the third adjusting screw 102 and the eighth threaded hole 102a of the positioning key 204 are used to adjust the length, the nut of the second adjusting screw 103 moves the second bottom plate 201 to the Y direction to achieve the adjusting purpose.

[0043] The third adjusting screw 102 is fixed with the eighth threaded hole 102a of the first fixed key 205, the nut of the adjusting screw is clamped in the first nut slot 102b on the second fixed key 206 of the third bottom plate 200, when the third direction freedom (X direction) needs to be adjusted, the third adjusting screw 102 and the third threaded hole 102a of the first fixed key 205 are used to adjust the length, the nut of the third adjusting screw 102 moves the third bottom plate 200 to the X direction to achieve the adjusting purpose.

[0044] The technical features of the above-mentioned embodiments can be combined in any way, in order to make the description simple, not all possible combinations of technical features in the above-mentioned embodiments are described, however, as long as the combination of technical features does not exist contradictory, it should be considered as the scope of the present application.

[0045] The above-mentioned embodiments are only the embodiments of the present application, and do not limit the patent scope of the present application, any equivalent structure or equivalent process transformation, or direct or indirect application in other related technical fields, are also included in the patent protection scope of the present application.

Claims

1. An optical system for a genetic sequencer, the optical system comprising: The application relates to a fluorescence imaging device. The device comprises a sample module, an illumination module, a focusing module, a light splitting module and an imaging module. The sample module comprises a sample. The illumination module comprises a light source, a lens group, a laser dichroic mirror, a laser filter and an objective lens. The focusing module comprises a focusing module group, a focusing dichroic mirror and a focusing filter. The light splitting module comprises a dichroic mirror group. The imaging module comprises a tube lens, an image sensor, an image correction device and a filter. The light source is used for emitting excitation light for exciting the sample, the lens group comprises a collimating mirror and an imaging lens, the collimating mirror is arranged in a light path between the light source and the laser dichroic mirror, the collimating mirror is used for expanding the light source excitation light, the imaging lens is arranged in the light path between the light source and the laser dichroic mirror, the imaging lens is used for imaging the expanded light source excitation light on a back focal plane of the objective lens, the laser dichroic mirror is arranged in a light path between the objective lens and the tube lens, the laser dichroic mirror is used for reflecting the light source excitation light into the objective lens to excite the sample and transmitting the emission light emitted from the sample into the image sensor, the laser filter is arranged in the light path between the light source and the laser dichroic mirror, the laser filter is used for ensuring the cleanliness of the light source excitation light, and the objective lens is used for collecting the excitation light for exciting the sample to the sample and collecting the emission light from the sample to the imaging module.

2. The optical system for a genetic sequencer of claim 1, wherein: The sample module comprises a sample with fluorescently labeled base sequence information, the sample can be excited by the light source to emit fluorescent signal emission light of different wavelengths, the fluorescent signal emission light enters the imaging module after passing through the light splitting module, and the image sensor in the imaging module captures the base sequence information.

3. The optical system for a genetic sequencer of claim 1, wherein: The focusing module group emits focusing laser, the focusing filter is arranged in a light path between the focusing module group and the focusing dichroic mirror, the focusing filter is used for ensuring the cleanliness of the focusing laser, the focusing dichroic mirror is arranged in a light path between the objective lens and the tube lens, the focusing dichroic mirror is used for reflecting the focusing laser into the objective lens to the sample, adjusting the vertical relative position of the objective lens and the sample, the sample reflects the focusing laser back to the focusing module to determine the vertical relative position of the objective lens and the sample, so that the focusing purpose is achieved.

4. The optical system for a genetic sequencer of claim 1, wherein: The light splitting module comprises a dichroic mirror group, the dichroic mirror group comprises a first dichroic mirror, a second dichroic mirror, a third dichroic mirror and a filter used for ensuring the cleanliness of the specific emission light from the sample, the dichroic mirror group is arranged in a light path between the excitation light from the light source to the objective lens and in a light path between the emission light from the objective lens to the tube lens, and the dichroic mirror is used for transmitting one of the excitation light and the emission light and reflecting the other one of the excitation light and the emission light.

5. The optical system for a genetic sequencer of claim 1, wherein: The imaging module comprises an image sensor for imaging the emitted light from the sample and a tube lens arranged to collect the emitted light from the objective to the image sensor, wherein the objective and the tube lens form an infinity conjugate, the image correction device comprises an image sensor adjustment device and a reflective adjustment mirror, the image sensor is mounted on the image sensor adjustment device, the reflective adjustment mirror is also used for image correction, the color filter is arranged in front of the image sensor, and the color filter ensures the cleanliness of the fluorescent signal emitted light.

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