CN207908817U - Lighting system - Google Patents

Lighting system Download PDF

Info

Publication number
CN207908817U
CN207908817U CN201721329355.0U CN201721329355U CN207908817U CN 207908817 U CN207908817 U CN 207908817U CN 201721329355 U CN201721329355 U CN 201721329355U CN 207908817 U CN207908817 U CN 207908817U
Authority
CN
China
Prior art keywords
light beam
speculum
lighting system
capture device
light
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN201721329355.0U
Other languages
Chinese (zh)
Inventor
M·陈
B·史
T·H·多
S·姚
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Life Technologies Holdings Pty Ltd
Original Assignee
Life Technologies Holdings Pty Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Life Technologies Holdings Pty Ltd filed Critical Life Technologies Holdings Pty Ltd
Application granted granted Critical
Publication of CN207908817U publication Critical patent/CN207908817U/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B27/00—Optical systems or apparatus not provided for by any of the groups G02B1/00 - G02B26/00, G02B30/00
    • G02B27/10—Beam splitting or combining systems
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B17/00—Systems with reflecting surfaces, with or without refracting elements
    • G02B17/02—Catoptric systems, e.g. image erecting and reversing system
    • G02B17/06—Catoptric systems, e.g. image erecting and reversing system using mirrors only, i.e. having only one curved mirror
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B27/00—Optical systems or apparatus not provided for by any of the groups G02B1/00 - G02B26/00, G02B30/00
    • G02B27/0025—Optical systems or apparatus not provided for by any of the groups G02B1/00 - G02B26/00, G02B30/00 for optical correction, e.g. distorsion, aberration

Landscapes

  • Physics & Mathematics (AREA)
  • General Physics & Mathematics (AREA)
  • Optics & Photonics (AREA)
  • Microscoopes, Condenser (AREA)

Abstract

The utility model provides a kind of lighting system, and the lighting system includes being configured to by imageable target surface placed on it, light source, beam splitter, at least further includes the first speculum.The beam splitter is configured to separate from the light beam of the light source, and first mirror arrangement is that will be reflected into from the first beam of the beam splitter on the surface with the imageable target.Imaging system includes being configured to imaging surface, speculum and capture device with the imageable target being disposed thereon.The capture device is configured to the image by extending from the imageable target, reflecting the speculum and reach imageable target described in the trace capture for emitting light of the capture device.The speculum, described capture device or both are each configured to move the length in the path to shorten the transmitting light in the diagonal directions relative to the imaging surface.It also discloses for calibrating imaging system to eliminate or reduce the heterogeneity caused by imaging system characteristic in sample image.

Description

Lighting system
Technical field
This disclosure relates to equipment, system and the side of the high-definition picture for object to be illuminated and obtained to object Method.Present disclosure also relates to the methods of image non-uniformity correction.
Background technology
The method independent of such as number amplification or using zoom lens is needed to provide the high-definition picture of object Imaging device, System and method for.As image is amplified, number amplification typically results in image pixel.It is difficult in many cases To use zoom lens, because being generally difficult to meet in a stable manner such as large aperture, focal length, operating distance, deformation, the curvature of field The various requirements such as rate and signal decaying.
It there is a need for being able to use when different and provide two beams or more Shu Guang in the case of two or more light sources with right Lighting apparatus that imageable target is illuminated, system and method, especially in uniform means of illumination.It is logical using multiple light sources The light beam for often resulting in different luminous powers is applied in imageable target, because must individually be safeguarded when using two light sources, and And there may be different optical characteristics after manufacturing or configuring in equipment or system.In addition, usual using multiple light sources Cause the overall illumination of imageable target that there is higher heterogeneity, and lighting system also has higher mechanical complexity, Its possibility for then improving maintenance requirement and increasing non-uniform illumination.During imaging another FAQs (with Imaging pattern is unrelated) it is image non-uniform.For example, when same sample is placed in imaging surface or the different location in the visual field, even if Same sample emits identical signal, and respective image also can seem uneven based on position.This field needs solution image non- Even property.
Invention content
The invention discloses a kind of lighting systems.The lighting system include surface, light source, beam splitter, the first speculum and Second speculum.Surface configuration is with the imageable target being disposed thereon.Light source is configured to project light beam.Beam splitter configures For the light beam from light source is separated into the first light beam and the second light beam.First mirror arrangement is the first light beam of reflection to provide To the first light beam of the reflection that surface is illuminated.Second mirror arrangement is that the second light beam of reflection shines surface with providing Second light beam of bright reflection.
In another embodiment, lighting system includes surface, light source, beam splitter and the first speculum.Surface configuration is With the imageable target being disposed thereon.Light source is configured to project light beam.Beam splitter is configured to will the light beam from light source point From for the first light beam and the second light beam.Second light beam illuminates surface.First mirror arrangement is that reflection comes from beam splitter The first light beam to provide the first light beam of the reflection illuminated to surface.
Also disclose a kind of means of illumination.This method includes providing the surface with the imageable target being disposed thereon.It should Method further includes providing light beam by light source.This method further includes that the Shu Guang is separated into the first light beam and the second light beam.It should Method further includes being illuminated to surface.Illumination includes：(i) using the first speculum reflect the first light beam with generate to surface into First light beam of the reflection of row illumination, and (ii) reflect the second light beam using the second speculum and are illuminated to surface with generating Reflection the second light beam.
In another embodiment, means of illumination includes providing light beam by light source.This method further includes by the Shu Guang It is separated into the first light beam and the second light beam.This method further includes that the surface of opposed imageable target thereon is illuminated.Illumination Including using the first speculum to reflect the first light beam to generate the first light beam of the reflection illuminated to surface.From the Shu Guangzhong Separate the second light beam so that second light beam illuminates surface.
Also disclose a kind of imaging system.The imaging system includes imaging surface, speculum and capture device.Imaging surface It is configured to imageable target placed on it.Capture device be configured to by extend from imageable target, reflect speculum and to Up to the image of the trace capture imageable target of the transmitting light of capture device.Speculum, capture device or both be configured to relative to Imaging surface moves the length in the path to shorten transmitting light in the diagonal directions.
In another embodiment, imaging system includes that imaging surface, speculum, speculum axis, capture device, capture are set Standby axis and drive block.Imaging surface is configured to imageable target placed on it.Mirror arrangement is along speculum axis at first pair Linea angulata moves on direction.Capture device is configured to by extending from imageable target, reflecting speculum and reach capture device Transmitting light trace capture imageable target image.Capture device is configured to diagonally adjacent second along capture device axis It is mobile.Drive block transmits the movement between speculum and capture device, to make speculum and capture device while movement.
Also disclose a kind of imaging method.This method includes that imageable target is placed on imaging surface.This method further includes Capture device, speculum or both is set to be moved in the diagonal directions both with respect to imaging surface.This method further includes that use is caught It obtains equipment and is imaged mesh by extending, reflecting speculum from imageable target and reach the trace capture for emitting light of capture device Target image.
Also disclose a kind of illumination and imaging system.The system includes being configured to the imageable target with being disposed thereon Surface.Light source is configured to project light beam.Beam splitter is configured to the light beam from light source being separated into the first light beam and the second light Beam.First illuminated mirror is configured to the first light beam of reflection to provide the first light beam of the reflection illuminated to surface.Second illumination Mirror is configured to the second light beam of reflection to provide the second light beam of the reflection illuminated to surface.Capture device be configured to by from The image that imageable target extends, reflects transmitting mirror and reach the trace capture imageable target of capture device.Transmitting mirror, capture Equipment or both is configured to be moved in the diagonal directions relative to surface to shorten the length in path.
Also disclose a kind of illumination and imaging method.This method includes that imageable target is placed on surface.This method is also wrapped It includes from light source and projects light beam.This method further includes that the Shu Guang is separated into the first light beam and the second light beam.This method further includes Imageable target is illuminated.Illumination includes：(i) the first light beam is reflected using the first illuminated mirror to illuminate surface to generate Reflection the first light beam, and (ii) using the second illuminated mirror reflect the second light beam to generate the reflection illuminated to surface The second light beam.This method further includes being caught using capture device by extending, reflecting transmitting mirror from imageable target and reach Obtain the image of the trace capture imageable target of equipment.
The disclosure describes the method for generating the image for passing through Nonuniformity Correction in some embodiments.At some In embodiment, heterogeneity is shown as the identical signal for being measured at the different location in visual field, modified strong Spend the image of signal.
The asymmetric correction method of the disclosure can be applied to the image obtained from various samples, and the sample includes biology Sample, the biological sample include biomolecule such as protein, polypeptide, glycoprotein, modified protein, nucleic acid, DNA, RNA, carbon water Compound, lipid, lipopolysaccharides, biopolymer, by cell and tissue generate other metabolins and combination thereof.Biology Molecule or biological sample with biomolecule can be individually imaged or dispersible, positioning or embedded film, gel, filter paper, carry glass Piece, microplate or matrix such as polyacrylamide gel or nitrocellulose or PDVF films trace, Ago-Gel, agar plate, It is imaged in tissue culture plate or histotomy.The asymmetric correction method of the disclosure can be applied to from above-mentioned sample The image of any acquisition.
The asymmetric correction method of the disclosure can be applied to the image generated by the chemiluminescence variation of biological sample Or the image of the change in fluorescence generation by sample.The asymmetric correction method of the disclosure can be applied to by bioluminescence at The image that picture, transillumination or reflected light imaging generate.
In one embodiment, it generates and includes by the method for the image of Nonuniformity Correction：It calculates on imaging sensor The relative illumination of the imaging len of multiple pixels；Flat field correction matrix is generated based on the relative illumination；Capture or acquisition one or The image of multiple biological samples, wherein image have heterogeneity；And the image captured using flat field correction adjustment of matrix To generate the image by Nonuniformity Correction.
In one embodiment, generation flat field correction matrix includes inverted to relative illumination to generate flat field correction square Battle array.In some embodiments, the formula obtained calculating relative illumination is returned using by linearly or nonlinearly curve matching.The song Line can be an order polynomial, quadratic polynomial, cubic polynomial etc..It calculates flat field correction matrix and generates flat field correction matrix value.
The image captured using flat field correction adjustment of matrix includes the one or more biological samples that will be captured or acquire Image be multiplied by the value of flat field correction matrix.In some embodiments, it captures or acquires using flat field correction adjustment of matrix Image further includes that the image for the one or more biological samples that will be captured or be acquired based on pixel to pixel is multiplied by flat field correction square The value of battle array is to generate the image through flat field correction.In some embodiments, the image display through flat field correction is captured or is acquired The accuracy of the signal level of each image of one or more biological samples, and it is unrelated with its position in visual field.
In one embodiment, present disclose provides a kind of method generating the image by Nonuniformity Correction, the party Method includes：Calculate the relative illumination of the imaging len of multiple pixels on imaging sensor；It is inverted to relative illumination flat to generate Field correction matrix；The image of capture or acquisition biological sample；And the biological sample that will be captured or acquire based on pixel to pixel Image be multiplied by the value of flat field correction matrix to generate the image through flat field correction.
In one embodiment, the present disclosure describes the method for generating the image by Nonuniformity Correction, this method packets It includes：The image of capture or the one or more biological samples of acquisition, wherein image have heterogeneity；And utilize flat field correction square The image that battle array adjustment is captured or acquired is to generate the image by Nonuniformity Correction.Adjusting the image for capturing or acquiring can Include that the image of one or more biological samples of capture is multiplied by generate by the value of flat field correction matrix based on pixel to pixel Image through flat field correction.
In some embodiments, flat field correction matrix is in image device or imaging system.Flat field correction matrix is for making It is available with the user of image device.Flat field correction matrix value can be stored in imaging device.In some embodiments, flat field correction Matrix value can be stored in the software component or machine element of imaging device.Flat field correction matrix value is for using imaging system The user for carrying out Nonuniformity Correction is available.In some embodiments, flat field correction matrix is flat field correction principal matrix.
In one embodiment, the method for heteropical flat field correction matrix of the generation for correcting image includes：Meter Calculate the relative illumination of the imaging len of multiple pixels on imaging sensor；Flat field correction matrix is generated based on relative illumination, and Flat field correction matrix is normalized based on the maximum pixel intensity value in matrix.In one embodiment, flat field school is generated Positive matrices includes inverted to relative illumination and the value in matrix is normalized.For example, manufacturer or user produce Flat field correction matrix in case use in the future.
Be described below it is middle will part illustrate additional objects and advantages of this invention, and the objects and advantages will part from It is clear that it can be by putting into practice acquistion to the present invention in description.Objects and advantages of the present invention will be by means of appended power The element that is particularly pointed out in sharp claim and combination are realized and are obtained.
It should be understood that being generally described above and described in detail below being only exemplary and explanatory and be not intended to limit institute The invention of opinion.
Attached drawing schematic illustration
Attached drawing is incorporated to and forms part of this specification, this is explained to illustrate the present invention and be used for together with this explanation The principle of invention.
Fig. 1 and Fig. 2 shows the perspective views (having an X-rayed from different perspectives) according to the imaging system of one embodiment.
Fig. 3 and Fig. 4 shows the perspective view of the imaging system according to one embodiment, some components wherein in Fig. 1 and Fig. 2 It is omitted so that the axis of coupling access component is better shown.
Fig. 5, Fig. 6, Fig. 7 and Fig. 8 show the cross section that the imaging system of scaling is continuously improved according to one embodiment Side view.
Fig. 9 shows the light beam projected from the lighting module of imaging system according to one embodiment.
Figure 10 shows to work as movement of the center in the path for emitting light with transmitting mirror during scaling according to one embodiment When keeping on the mirror between two parties, primary beam is penetrated by falling for reflective mirror blocks at least partly.
Figure 11 shows the simplification of the light beam projected from the lighting module of imaging system shown in Fig. 9 according to one embodiment Schematic side elevation.
Figure 12 shows showing for the simplification of the light beam projected from the lighting module with additional reflector according to one embodiment Meaning property top view.
Figure 13 shows the showing from the simplification for having the light beam projected there are two the lighting module of beam splitter according to one embodiment Meaning property top view.
Figure 14 shows cross-sectional side view of the light beam according to one embodiment before reaching beam splitter across hole.
Figure 15 shows the perspective view of photometer reference plate.
Figure 16 A show the stacking image of a luminous point of the photometer reference plate at the different location of visual field, and scheme 16B illustrates that the figure of the signal of same spot at different location.
Figure 17 shows to indicate the relative illumination of the imaging len in imaging system according to one embodiment and by non-linear Return obtained best fit result.
Figure 18 A show to pass through the modified figure of Figure 16 A obtained using flat field correction principal matrix according to one embodiment Picture, and Figure 18 B illustrate that the figure of the ratio of the intensity of spot and the maximum intensity of spot.
Figure 19 A show that the image of the chemiluminescence sample among visual field under 1 times of scaling, Figure 19 B are shown The figure of the chemiluminescence sample at position under 1 times of scaling between the centre position of visual field and upper right diagonal position Picture, and 19C shows the image of the chemiluminescence sample under 1 times of scaling at the upper right diagonal position in visual field.
Figure 20 A show that the image according to the progress quantitative analysis of two row band of one embodiment pair, Figure 20 B show Figure 20 A's The figure of intensity of the first row band before flat field correction, Figure 20 C show the first row band of Figure 20 A after flat field correction The figure of intensity, Figure 20 D show the figure of intensity of the second row band of Figure 20 A before flat field correction, and Figure 20 E show figure The figure of intensity of the second row band of 20A after flat field correction.
Figure 21 A show the relative illumination according to one embodiment imaging len when amplifying 1 times to sensor image height Table, and Figure 21 B show the relative illumination of the imaging len when amplifying 2 times to the table of sensor image height.
Figure 22 shows the curve graph relative to the centrosymmetric relative illuminations of CCD according to one embodiment.
Figure 23 A show the figure according to one embodiment best fit nonlinear regression curve when amplifying 1 times, and scheme 23B shows the figure of the best fit nonlinear regression curve when amplifying 2 times.
Figure 24 shows the analog image when amplifying 1 times according to one embodiment.
Figure 25 shows the flat field correction main picture according to one embodiment.
Figure 26 A, Figure 26 B and Figure 26 C are using the image after flat field correction.Figure 26 A show put according to one embodiment The image in the centre position in visual field at big 1 times, Figure 26 B are shown when amplifying 2 times between the centre position of visual field and upper right The image at position between diagonal position, and Figure 26 C are shown when amplifying 1 times at the upper right diagonal position in visual field Image.
Figure 27 shows the image of each eight bands of two rows according to one embodiment.
Figure 28 A show figure of the first row according to one embodiment Figure 27 when amplifying 1 times before flat field correction, figure 28B shows figure of the first row of Figure 27 when amplifying 1 times after flat field correction, and Figure 28 C show Figure 27 when amplifying 1 times Figure of second row before flat field correction, and Figure 28 D show the second row of Figure 27 when amplifying 1 times after flat field correction Figure.
Figure 29 A show figure of the first row according to one embodiment Figure 27 when amplifying 2 times before flat field correction, figure 29B shows figure of the first row of Figure 27 when amplifying 2 times after flat field correction, and Figure 29 C show Figure 27 when amplifying 2 times Figure of second row before flat field correction, and Figure 29 D show the second row of Figure 27 when amplifying 2 times after flat field correction Figure.
Figure 30 shows the film location (for example, intermediate, upper right corner) according to one embodiment before and after flat field correction Chart.
Specific implementation mode
With detailed reference to the exemplary embodiment of the present invention, illustrate the reality of the exemplary embodiment in the accompanying drawings Example.In the conceived case, it will pass through schema and refer to same or similar component using identical reference label.It is retouched following In stating, with reference to forming part thereof of attached drawing, and wherein by means of explanation, that wherein the present invention may be practiced is specific exemplary real It applies and exemplifies.Fully describe these embodiments in detail, so that those skilled in the art can put into practice the present invention, and And it should be understood that using other embodiments and change can be made without departing substantially from the scope of the present invention.Therefore, below It describes exemplary only.
Fig. 1 and Fig. 2 shows the perspective views of the part of the imaging system 100 obtained from different perspectives according to one embodiment. Imaging system 100 may include imaging surface 110.In an example, imaging surface 110 can be or including pallet or screen. Imaging surface 110 can be plane and substantially horizontal (that is, being parallel to ground).Imageable target 112 can be placed in into On image surface 110.Imageable target 112 can be or including with polyacrylamide gel, Ago-Gel, nitrocellulose filter and The relevant biomaterial of pvdf membrane such as nucleic acid and/or protein.Imageable target 112 can also be or all including non-biological material Such as the product and document of manufacture.
Imaging system 100 may also include speculum 120.Speculum 120 can be positioned in the imaging surface (for example, directly) 110 and the top of imageable target 112.Speculum 120 may include reflecting surface.As shown, reflecting surface can be plane；So And in other embodiments, reflecting surface can be bending.When the reflecting surface of speculum 120 is plane, speculum 120 reflecting surface can be orientated angled relative to imaging surface 110 (that is, relative to horizontal direction).The angle can be with It is about 10 ° to about 80 °, about 20 ° to about 70 ° or about 30 ° to about 60 °.For example, the angle can be about 45 °.
Imaging system 100 may also include capture device 130.Capture device 130 may include detector housing 140, one or Multiple optical filters (one of them is as shown in 150) and camera 160.Detector housing 140 can be positioned in 110 top of imaging surface And laterally (for example, horizontal) shifted reflections mirror 120.Detector housing 140 may include lens 142.Detector housing 140 may be used also The sensor of focus and hole including filter wheel, motor and/or control lens 142.Lens 142 can be plane, and pass through The central longitudinal axis of lens 142 can intersect with the reflecting surface of middle speculum 120.As such, the path of transmitting light can be Extend in vertical direction between imageable target 112 and speculum 120, and in the speculum of detector housing 140 120 and lens It is extended transversely between 142.As used herein, " path of transmitting light " refers to passing through lens 142 from imageable target 112 in visual field To the path of camera 160.
Optical filter 150 can be couple to and be positioned at behind detector housing 140, and the path for emitting light can prolong It extends through detector housing 140 and enters in optical filter 150.Optical filter 150 can be only by the optical transport of selected wavelength to phase Electromagnetism (" EM ") optical filter of machine 160.Optical filter 150, which is placed in behind lens 142, allows optical filter 150 to be less than optical filter 150 the case where being placed in 142 front of lens.Exciting light and transmitting light can enter lens 142.Swashed by what optical filter 150 stopped Lens 142 and circumferential surface can be hit by shining, and a certain amount of exciting light can again return to optical filter 150 and can be at this moment Across optical filter 150.In another embodiment, optical filter can be placed in the front of lens 142.Due to exciting by lens The optical filter blocking in 142 fronts, therefore might have considerably less exciting light after optical filter, for example, almost without excitation In light propagation to lens 142 and camera 160 is reached, this makes it easy to control stray light and reduces background signal.Lens The optical filter in 142 fronts can be more than the subsequent optical filter of lens 150.Therefore, the size of filter wheel bigger and may occupy more Big space.In certain embodiments, the second optical filter can also be placed in 142 front of lens.In such embodiments, certain implementations Can be that the second optical filter of notch filter is placed in the front of lens 142, and optical filter 150 is placed in 142 rear of lens in example.This A little embodiments can provide the advantages of two filters work at the same time, utmostly to weaken including stray excitation Influence of the stray light to the transmitting captured by camera 160.
Camera 160 can be couple to and the path for being located in behind optical filter 150, and emitting light can extend across optical filtering In device 150 and entrance camera 160, wherein camera 160 can capture the one or more of imageable target 112 (for example, through filtering ) image.
Imaging system 100 may also include first sensor 190 at first position and positioned at the second place Second sensor 192 (is shown in FIG. 1).First sensor 190 can be limiting sensor, which is configured to limit The travel distance of detector housing 140, optical filter 150 and camera 160.Second sensor 192 can be target-seeking sensor, this is target-seeking Sensor is configured to set detector housing 140, optical filter 150 and camera 160 to initial default location.
Imaging system 100 may also include lighting module 200 (as shown in Figure 1).Lighting module 200 can be or be penetrated including falling Lighting module and/or transillumination module.Lighting module 200 may include light source 210.Light source 210 can be or including one or Multiple light emitting diodes (" LED ").Lighting module 200 may also include exciter filter 220, which is couple to And it is positioned at the front of light source 210.Exciter filter 220 can be configured to limit the wave-length coverage of the light from light source 210. Lighting module 200 may also include lens 230, which is couple to and is positioned at the front of exciter filter 220.Extremely In few one embodiment, lens 230 can be or including lens ring.Lighting module 200 may also include beam splitter 240, this point Beam device 240 is couple to and is positioned at the front of lens 230.Beam splitter 240 can be configured to separate from the light beam of light source 210 Or it is divided into two or more beam sections.Lighting module 200 may also include near-infrared (" NIR ") lighting module and can quilt Be positioned close to the speculum 250 of light source 210 (for example, being below), exciter filter 220, lens 230, beam splitter 240 or Combination thereof.NIR lighting modules and speculum 250 may include the LED for providing the light within the scope of NIR.NIR lighting modules and Speculum 250 can also be substantially the same with visible light angle NIR light is reflected into beam splitter 240.Lighting module 200 May also include the back mirror 260 for being positioned in 210 top of the lower section of capture device 130 and/or light source, exciter filter 220, Lens 230, beam splitter 240 or combination thereof.Lighting module 200 may also include front mirror 262.Imaging surface 110 can be horizontal Light source 210, exciter filter 220, lens 230, the side of beam splitter 240 and front-reflection are positioned at (for example, flatly) to ground Between the other side of mirror 262.Front mirror 262 can also be positioned in 110 top of imaging surface.Although being not shown, illumination Module 200 may also include transillumination module and light source (for example, LED).A light source or multiple light sources for transillumination 110 lower section of imaging surface can be positioned in provide illumination by imaging surface 110 and imageable target 112.
Fig. 3 and Fig. 4 shows the perspective view of the imaging system 100 according to one embodiment, and some of components are (for example, anti- Penetrate mirror 120 and capture device 130) it is omitted so that the axis 124,134 of coupling access component is better shown.
Speculum 120 (being not shown in figs. 3 and 4) can be couple to reflecting mirror support structure 122, and capture device 130 (being also not shown in figs. 3 and 4) can be couple to capture device support construction 132.Reflecting mirror support structure 122 can couple To and be configured to slide back and forth being aligned with speculum axis 124 in the axial direction of (for example, parallel) along speculum axis 124.Capture Equipment support construction 132 can be couple to and be configured to along capture device axis 134 be aligned with capture device axis 134 (for example, It is parallel) axial direction on slide back and forth.Drive block 180 can be couple to and be configured to along drive block axis 184 with drive block axis 184 It is aligned in the axial direction of (for example, parallel) and slides back and forth.In at least one embodiment, speculum axis 124, capture device axis 134, Drive block axis 184 or combination thereof can be in single plane.
Speculum axis 124 can be in diagonal orientation relative to the upper surface of imaging surface 110.As used herein, " diagonal " is Refer to direction that is both not parallel or being not orthogonal to imaging surface 110.More specifically, speculum axis 124 can be relative to imaging surface 110 are orientated angled, which is about 10 ° to about 170 °, about 40 ° to about 140 ° or about 70 ° to about 110 ° (when from Fig. 3 When being observed with direction shown in Fig. 4).It (is carried out when from Fig. 3 and angle shown in Fig. 4 for example, angle 126 can be about 91 ° When observation).
Capture device axis 134 also can be in diagonal orientation relative to imaging surface 110 (that is, relative to horizontal direction).More Body, capture device axis 134 can be orientated angled relative to imaging surface 110, which is about 10 ° to about 80 °, about 20 ° to about 70 ° or about 30 ° to about 60 ° (when from being carried out from direction shown in Fig. 3 and Fig. 4).For example, angle 136 can be with It is about 35 ° (when from being carried out from Fig. 3 and angle shown in Fig. 4).Angle between speculum axis 124 and capture device axis 134 Degree 127 can be about 80 ° to about 140 °, about 90 ° to about 130 ° or about 100 ° to about 120 °.For example, angle 127 can be About 123 °.
Drive block axis 184 can be positioned between speculum axis 124 and capture device axis 134 (that is, being in angle 127 It is interior).Drive block axis 184 can be also orientated relative to the upper surface of imaging surface 110 in diagonal orientation or vertical (that is, vertical).
Referring to Fig. 4, the first transmission shaft 138 can be couple to and capture device support construction 132 and drive block 180 it Between extend.Capture device support construction 132 (and capture device 130), drive block 180 or combination thereof can be configured to along first Transmission shaft 138 slides axially.Second driving shaft 128 can be couple to and between reflecting mirror support structure 122 and drive block 180 Extend.Reflecting mirror support structure 122 (and speculum 120), drive block 180 or combination thereof can be configured to along the first transmission shaft 128 slide axially.
Imaging system 100 may include one or more motors (one of them is as shown in 170 in Fig. 3).Motor 170 can make Speculum 120 and/or capture device 130 (for example, detector housing 140, optical filter 150 and camera 160) are obtained relative to imaging Surface 110 and imageable target 112 move.In the illustrated embodiment, single motor 170 may make speculum 120 and capture to set Standby 130 is mobile simultaneously.This while movement can be realized by using power transmission shaft and drive block, the power transmission shaft and Drive block couples speculum 120 and capture device 130, is such as driven above in association with the first transmission shaft 138, second described in Fig. 4 Axis 128 and drive block 180.Such method provides the movement that speculum 120 and capture device 130 are controlled using single motor Advantage, and controlled motion is come with the method for synchronization independent of the independent control mechanism such as control software, to provide drop Low-complexity and cost reduce the advantages of maintenance needs, and improve in the image being consistent under different zoom degree The ability of the heart.In another embodiment, first motor may make speculum 120 to move, and the second motor may make capture Equipment 130 moves, and speculum 120 can be fixed relative to the ratio of the movement of capture device 130.Software can be passed through First motor and the second motor are controlled to realize this fixed movement ratio, and can realize and move synchronously, while being contracted Keep the center of image consistent during putting.Drive block 180 can be couple to speculum 120 and capture device 130.When using single electric When machine 170, drive block 180 can couple the movement of speculum 120 and capture device 130, as Fig. 3 and Fig. 4 are described in detail.Other In embodiment, one or more belt drives or miscellaneous equipment can be used for mobile mirror 120 and capture device 130.
Referring again to Fig. 3 and Fig. 4, motor 170 can be couple to lead screw 172 by coupler 174.Coupler 174 can incite somebody to action The rotary motion of motor 170 is transferred to lead screw 172, to make lead screw 172 rotate.Lead screw 172 can be with capture device axis 134 It is parallel.When lead screw 172 is rotated in a first direction, lead screw 172 can push up capture along capture device axis 134 in first axle Equipment support construction 132 (and capture device 130).On the contrary, when (that is, opposite) direction rotates lead screw 172 along second, silk Bar 172 can (that is, opposite) side pulls up capture device support construction 132 and (and catches second along capture device axis 134 Obtain equipment 130).
When capture device support construction 132 (and capture device 130) is moved up along capture device axis 134 in first axle When, the first transmission shaft 138 can be such that drive block 180 is moved up in first axle along drive block axis 184.On the contrary, when capture is set For standby support construction 132 (and capture device 130) along capture device axis 134 when the second axis moves up, the first transmission shaft 184 can So that along drive block axis 184, (that is, opposite) axis moves up drive block 180 second.
When drive block 180 along drive block axis 184 when first axle moves up, second driving shaft 128 can make speculum Support construction 122 (and speculum 120) is moved up along speculum axis 124 in first axle.On the contrary, when drive block 180 is along biography For motion block axis 184 when the second axis moves up, second driving shaft 128 can make reflecting mirror support structure 122 (and speculum 120) Along speculum axis 124, (that is, opposite) axis is moved up second.
It will thus be appreciated that speculum 120 and capture device 130 can move together simultaneously.When speculum 120 and catch When obtaining equipment 130 and being moved along its corresponding first axis, from imageable target 112 (reflecting speculum 120) to detector housing The total length in the path of the transmitting light of 140 lens 142 can reduce, and when speculum 120 and capture device 130 are along its phase When second answered is axially moved, from imageable target 112 (reflecting speculum 120) to the hair of the lens 142 of detector housing 140 Penetrating the total length in the path of light can increase.
Fig. 5, Fig. 6, Fig. 7 and Fig. 8 show the cross that the imaging system 100 of scaling is continuously improved according to one embodiment Side cross-sectional view.More specifically, Fig. 5 shows the imaging system not scaled 100.In one embodiment, when not scaling, from As target 112 (reflecting speculum 120) arrives the total of 115 center of path of the transmitting light of the lens 142 of detector housing 140 Length can be for example, about 455mm, but the total length for emitting 115 center of path of light will be depending on the total of system and its component Body configures, and includes the characteristic of capture device 130.When not scaling, the path 114 for emitting light can be with first of speculum 120 Divide (for example, surface region) contact.First part's (for example, surface region) can be the pact in the total surface region of speculum 120 50% to about 100%, about 75% to about 99% or about 85% to about 95%.
Referring now to Fig. 6, capture device 130 and speculum 120 can be moved along its corresponding first axis to reduce hair Penetrate the total length (that is, imageable target 112 on amplification imaging surface 110) at 115 center of path of light.Between imageable target 112 The center in the path 115 of the transmitting light between speculum 120 can be kept fixed in 120 diagonal motion of speculum (that is, Vertical arrows are identical with Fig. 5 and Fig. 6).Therefore, the point for emitting the speculum of the central contact in the path 115 of light can be with anti- Mirror 120 and capture device 130 is penetrated to move along its corresponding first axis and change/move.For example, the path 115 of transmitting light Center can be in Figure 5 point 116A at and point 116B in figure 6 at contacted with speculum 120.In addition, the road of transmitting light The part (for example, surface region) for the speculum 120 that diameter 114 contacts can with speculum 120 and capture device 130 along its first It is axially moved and reduces.
Referring now to Fig. 7, speculum 120 and capture device 130 can also be moved along its corresponding first axis with into one Step reduces the total length (that is, imageable target 112 on amplification imaging surface 110) at 115 center of path of transmitting light.Between imaging The center in the path 115 of the transmitting light between target 112 and speculum 120 can keep solid in 120 diagonal motion of speculum Fixed (that is, vertical arrows are identical with Fig. 5-7).Therefore, the point for emitting the speculum of the central contact in the path 114 of light can be with Speculum 120 and capture device 130 move along its corresponding first axis and change/move.For example, the path 114 of transmitting light Center can be in the figure 7 point 116C at contacted with speculum 120.In addition, the speculum that the path 114 of transmitting light contacts 120 part (for example, surface region) can further be moved along its first axis with speculum 120 and capture device 130 and Continue to reduce.
Referring now to Fig. 8, speculum 120 and capture device 130 can utmostly reduce 115 center of path for emitting light Total length (that is, utmostly amplify imaging surface 110 on imageable target 112).Between imageable target 112 and speculum The center in the path 115 of the transmitting light between 120 can be kept fixed in 120 diagonal motion of speculum (that is, vertical arrows with It is identical in Fig. 5-8).Therefore, the point for emitting the speculum of the central contact in the path 115 of light can be with speculum 120 and capture Equipment 130 moves along its corresponding first axis and changes/move.For example, the center in the path 115 of transmitting light can be in Fig. 8 In point 116D at contacted with speculum 120.In an example, when amplification factor maximum, from (anti-from imageable target 112 Project speculum 120) can be to the total length at center in path 115 of transmitting light of lens 142 of detector housing 140 For example, about 215mm.Therefore, imaging system 100 can be configured to be amplified to about 2 times from about 1 times；However, in other embodiments, Imaging system 100 can be set to be further amplified (that is, being more than 2 times).In addition, the speculum that the path 114 of transmitting light contacts 120 part (for example, surface region) can increase with amplification factor and be reduced.For example, when amplification factor maximum, the part (for example, surface region) can be speculum 120 total surface region about 5% to about 80%, about 10% to about 70% or About 20% to about 60%.
Fig. 9 shows the light beam (penetrating primary beam for example, falling) 212 projected from lighting module 200 according to one embodiment. Light beam 212 can be projected from the light source 210 (see Fig. 1) of lighting module 200.Light beam 212 can be separated into the first light beam by beam splitter 240 213 and second light beam 214.First light beam 213 can reflect back mirror 260 and be illuminated to imageable target 112, and And second light beam 214 can reflect front mirror 262 and imageable target 112 is illuminated.This gives below in conjunction with Figure 11 With more detailed description.In another embodiment, the NIR that light beam 212 can be projected and be reflected from NIR lighting modules 250 Speculum in lighting module 250 and reach imageable target 112.In at least one embodiment, light beam 212 can extend across hair The path 114 of light is penetrated to be illuminated to imageable target 112, with indirect illumination light or can be able to include to be swashed by falling to penetrate to illuminate The fluorescence part of light is sent out after hair.
When speculum 120 and capture device 130 are in the position of its maximum amplification, as shown in figure 9, capture device 130 lower end 139 can be positioned on the lower section of 120 lower end 129 of speculum.Therefore, speculum 120 is in appointing along speculum axis 124 It can not stop light beam 214 at meaning point.
Figure 10 shows to penetrate primary beam 212 by the falling of stopping of speculum 120 at least partly according to one embodiment.If hair The center for penetrating the path 114 of light remains fixed in same point on speculum 120 when speculum moves (for example, in Fig. 5 Point 116A), then when capture device 130 and speculum 120 are in the position of its maximum amplification, the lower end of speculum 120 129 can be positioned in the lower section of 130 lower end 139 of capture device.Therefore, speculum 120 can stop light beam 212 at least partly. Thus, as shown in figures 5-9, emit the path 114 of light center can with speculum 120 move and moved on speculum 120/ Change in order to avoid stopping light beam 212.
It falls and penetrates illumination and/or excitation can be used for the fluorescence mode of protein analysis.Many fluorescent dyes can be used for protein Dyeing and/or Western blotting, and there is different dyes different excitation spectrums to be distributed, it is therefore desirable to the excitation of different colours Light.Certain exciting powers can provide fluorescence imaging signal within the acceptable Imagewise exposure time.If illumination and/or excitation work( Rate changes too greatly in visual field, then there may be one or more dark spaces, wherein it is difficult to see that line band/band of sample, or can To see line band/band in dark space, but the signal of brighter areas becomes to be saturated.Therefore, substantially uniform illumination can change Kind image quality.
It falls there are two classes and penetrates illumination：Axial and off-axis (that is, inclination).Axial illumination can generate speck on the image, because Certain light reflections are from sample.Off-axis illumination is a kind of method for coping with this problem.In some embodiments, off-axis angle can be big In or equal to predetermined quantities in order to avoid generating speck.
Figure 11 shows the signal of the simplification of the light beam 212 projected from lighting module 200 shown in Fig. 9 according to one embodiment Property side view.Light beam 212 can be projected from the light source 210 (see Fig. 1) of lighting module 200.Light source 210 may include being used for fluorescence excitation The first LED and the 2nd LED for near-infrared.In another embodiment, usable halogen tungsten lamp covers two kinds of spectrum.For Any specific channel, can there is only light beams.Light source 210 can have solid color.In at least one embodiment, light Source 210 can be white light source, and optical filter can be used for generating different colors.
Light beam 212 can be separated into the first light beam 213 and the second light beam 214 by beam splitter 240.Although being not shown, In other embodiments, beam splitter 240 can be configured to light beam 212 being separated into three beams or more beam.As used herein, term " beam splitter " includes the one or more optical components that can separate or other means separation light beam, and including but not limited to rib Mirror, plate, dielectric mirror, metal coating speculum, beam splitter cube, fiber optic splitter and be configured to generate two or Light is set to collimate bundles of optical fiber before more output beams.
Beam splitter 240 can averagely be separated between the light beam of gained or peeling strength, or can they be separated into difference The intensity of ratio.In the illustrated embodiment, beam splitter 240 is plate, and the first light beam 213 reflects beam splitter 240, and the Two light beams 214 pass through beam splitter 240.Beam splitter 240 may include coating and/or optical filter (for example, linear variable filter), So that one end/side of beam splitter 240 can have the characteristic different from opposed end/side.First light beam 213 may include beam 212 Luminous power about 40% to about 60% (for example, 40%, 45%, 50%, 55% or 60%), and the second light beam 214 can wrap Include about 40% to about 60% (for example, 40%, 45%, 50%, 55% or 60%) of the luminous power of beam 212.Therefore, certain In embodiment, the first light beam 213 and the second light beam 214 can equably separate the luminous power of beam 212 (the first light beam 213 account for 50% And 50%) the second light beam 214 accounts for.In other embodiments, the first light beam 213 can have beam compared to the second light beam 214 The greater or lesser percentage of 212 luminous power.Angle between the center of first light beam 213 and the center of the second light beam 214 Degree can be about 62 ° to about 68 °, about 70 ° to about 90 ° or about 90 ° to about 110 °.First light beam 213 can reflect back mirror 260, generate the first light beam 215 of the reflection illuminated to the imageable target 112 on imaging surface 110.Second light beam 214 can Front mirror 262 is reflected, the second light beam of the reflection illuminated to the imageable target 112 on imaging surface 110 is generated 216.Angle between the center of first light beam 215 of reflection and the center of the second light beam 216 of reflection can be about 80 ° to about 100 °, about 106 ° to about 114 ° or about 120 ° to about 140 °.Although being not shown, at least one embodiment, second Light beam 214 can directly illuminate the imageable target 112 on imaging surface 110, without reflecting front mirror 262 and generating Second light beam 216 of reflection.
First light beam 215 of reflection and the second light beam 216 of reflection can be provided to the imageable target on imaging surface 110 112 off-axis illumination.More specifically, the first light beam 215 of reflection and the second light beam 216 of reflection can be provided to imaging surface The illumination of the substantial symmetry of imageable target 112 on 110.For example, between the first light beam 215 and imaging surface 110 of reflection Angle can be at reflection the second light beam 216 and imaging surface 110 between +/- 10 ° of angle within.From beam splitter 240 to The distance of back mirror 260 to imaging surface 110 is substantially equal to (for example, difference 10% within) from beam splitter 240 to preceding Speculum 262 arrives the distance of imaging surface 110.In at least one embodiment, back mirror 260 and/or front mirror 262 Movement can be combined with the rotation of beam splitter 240, to change the illumination to imageable target 112 on imaging surface 110.
Figure 12 is shown according to one embodiment from the injection of lighting module 1200 with additional reflector 1261-1265 The schematic top view of the simplification of light beam 1212.In the illustrated embodiment, light beam 1212 can be from the light source of lighting module 1200 It projects, and the first light beam 1213 and the second light beam 1214 can be separated by beam splitter 1240.First light beam 1213 can reflect Beam splitter 1240, and the second light beam 1214 may pass through beam splitter 1240.First light beam 1213 is can be on imaging surface 110 Imageable target 112 carry out illumination front-reflection go out the first speculum 1261 and the second speculum 1262.Second light beam 1214 is can Illumination front-reflection is carried out to the imageable target 112 on imaging surface 110 and goes out third speculum 1263, the 4th speculum 1264 and the Five speculums 1265.Identical as the embodiment in Figure 11, beam 1213,1214 can be provided to the imageable target on imaging surface 110 112 off-axis illumination.In addition, beam 1213,1214 can provide the substantial symmetry to the imageable target 112 on imaging surface 110 Illumination.
Figure 13 shows that there are two the injections of the lighting module 1300 of beam splitter 1340,1342 from tool according to one embodiment The schematic top view of the simplification of light beam 1312.In the illustrated embodiment, light beam 1312 can be from the light source of lighting module 1300 It projects, and can the first light beam 1313 and the second light beam 1314 be separated by the first beam splitter 1340.First light beam 1313 can be anti- The first beam splitter 1340 is projected, and the second light beam 1314 may pass through the first beam splitter 1340.In at least one embodiment, first Light beam 1313 may include the luminous power of beam 1312 about 15% to about 35% (for example, 15%, 20%, 25%, 30% or 35%), and the second light beam 1314 may include beam 1312 luminous power about 65% to about 85% (for example, 65%, 70%, 75%, 80% or 85%).
Then, the first light beam 1313 can reflect the first speculum 1360, generate to the imageable target on imaging surface 110 First light beam 1315 of 112 reflections illuminated.Second light beam 1314 can be separated into third beam by the second beam splitter 1342 1316 and the 4th beam 1317.Third beam 1316 can reflect the second beam splitter 1342, and the 4th beam 1317 may pass through the second beam splitting Device 1342.In at least one embodiment, third beam 1316 may include about 20% to about 40% (example of the luminous power of beam 1314 Such as, 33%), and the 4th beam 1317 may include beam 1314 luminous power about 60% to about 80% (for example, 66%).Then, Third beam 1316 can reflect the second speculum 1362, and generation illuminates the imageable target 112 on imaging surface 110 anti- The third beam 1318 penetrated.Identical as the embodiment in Figure 11, beam 1315,1318 can be provided to the imaging mesh on imaging surface 110 The off-axis illumination of mark 112.In addition, beam 1315,1318 can be provided to the substantially right of the imageable target 112 on imaging surface 110 The illumination of title.
4th beam 1317 can also illuminate the imageable target 112 on imaging surface 110.As shown, the 4th beam 1317 can not reflect speculum before shining the imageable target 112 on imaging surface 110.In one embodiment In, the angle between the first light beam 1313 and the 4th beam 1317 can be at the angle between third beam 1316 and the 4th beam 1317 Within about 10 ° to about 40 °.Similarly, in one embodiment, the angle between the first light beam 1315 and the 4th beam 1317 of reflection Degree can be within about 10 ° to about 40 ° of the angle between the third beam 1318 and the 4th beam 1317 of reflection.
Although Figure 13 show to the imageable target 112 on imaging surface 110 is illuminated beam 1315,1317, 1318, but in another embodiment, four or more beams can shine the imageable target 112 on imaging surface 110 It is bright.For example, four fronts Shu Kecong, rear portion, left and right side illuminate the imageable target 112 on imaging surface 110.
Figure 14 shows to be reached across the hole of beam shaping 1,410 1418 according to the light beam 212 in one embodiment Fig. 9 Cross-sectional side view before beam splitter 240.Beam shaping 1410 may include one or more lens, and (three lens are such as 1412, shown in 1414,1416).As shown, hole 1418 can be positioned between the second lens 1414 and the third lens 1416； However, in other embodiments, any position or divide in light arrival that hole 1418 can be positioned in beam shaping 1410 The outside of beam shaping 1410 is positioned in before beam device.The size (for example, cross-sectional area or diameter) in hole 1418 can be It is fixed.In another embodiment, the size in hole 1418 can be changed to change the photograph of imageable target 112 on imaging surface 110 Bright size (for example, cross-sectional area or diameter).The intensity of the also changeable light beam 212 from light source is to change imaging surface 110 The illumination intensity of upper imageable target 112.
Flat field correction is calibrated
The image device or imaging system of the disclosure can be used for being imaged various biomolecule and biological sample, described Biomolecule and biological sample such as protein, polypeptide, glycoprotein, modified protein, nucleic acid, DNA, RNA, carbohydrate, fat Matter, lipopolysaccharides, biopolymer and the other metabolins generated by cell and tissue.Biological sample can be individually imaged, or It can be at film, gel, filter paper, glass slide, microplate or matrix such as polyacrylamide gel or nitrocellulose or PDVF films print It is imaged in mark, Ago-Gel, agar plate, tissue culture plate or histotomy.
The imaging system of the disclosure can be imaged biomolecule and biological sample under several imaging patterns, including glimmering Light imaging, chemiluminescence imaging, biodiversity resources, transillumination or reflected light imaging.In some imaging patterns, sample shine or The variation (wavelength, frequency or Strength Changes) for showing the light of transmitting can be imaged without exterior lighting or excitation.In some imagings In pattern, sample shines or the light (wavelength, frequency or Strength Changes) of change transmitting, is then exposed to exterior lighting or excitaton source It is imaged.In some imaging patterns, sample reflected light or the light (wavelength, frequency or Strength Changes) for changing reflection, then Illumination is externally exposed to be imaged.
Faced in imaging process one common problem encountered is that (unrelated with imaging pattern), when sample be placed in imaging surface or When the different location of visual field, the different location of image seems uneven.Imaging surface is in one embodiment by the portion in Fig. 1 Part 110 is exemplified, and referred to herein as imaging region, visual field, sample screen or sample tray.In some embodiments In, image non-uniform is shown as, for the identical signal measured at the different location on imaging surface or visual field, having The image of the strength signal of variation.In some embodiments, image non-uniform is shown as in imaging surface or visual field On different location at the identical signal that measures, the image with unlike signal level.Image non-uniform related with position A kind of characteristic i.e. relative illumination for being partly due to imaging len causes.
The heterogeneity of the image of position based on sample on imaging surface is interfered carries out accurate quantitative analysis to biomolecule It measures.The present disclosure describes system, algorithm and the method for the lens subassembly for calibrating imaging system, the system, algorithm and sides Method can eliminate the heterogeneity that lens are shown to obtain accurate data from the sample image of biomolecule.Utilize the disclosure Method and system calibration lens subassembly eliminates the heterogeneity that lens are shown to obtain accurate data from sample image.
As described in foregoing embodiments, the imaging of the disclosure and lighting apparatus provide image correction function to pass through with system Image analysis improves the data precision.These functions are individually or with method and system further combined with non-homogeneous to calibrate image Property, to provide the excellent and accurate quantitative measurment to biological sample in running gel or film, and then provide reliable Data analysis and image information derived from sample.
In one embodiment, it generates and includes by the method for the image of Nonuniformity Correction：It calculates on imaging sensor The relative illumination of the imaging len of multiple pixels；Flat field correction matrix is generated based on the relative illumination；Capture is one or more raw The image of object sample, wherein image have heterogeneity；And using the image that flat field correction adjustment of matrix is captured to generate By the image of Nonuniformity Correction.
In one embodiment, the image captured using flat field correction adjustment of matrix includes that will be caught based on pixel to pixel The image of the biological sample obtained is multiplied by the value of flat field correction matrix to generate the image through flat field correction.
In some embodiments, a kind of generate may include by the method for the image of Nonuniformity Correction：Imaging is calculated to pass The relative illumination of the imaging len of multiple pixels on sensor；It is inverted to relative illumination to generate flat field correction matrix；By flat field Correction matrix is supplied to user；Wherein the image of the biological sample of capture can be multiplied by flat field correction by user based on pixel to pixel The value of matrix is to generate the image through flat field correction.User is handling it using flat field correction matrix to image progress flat field correction Preceding usable image device obtains the image of capture.In some embodiments, user, which may be selected to generate, passes through Nonuniformity Correction Image.It in some embodiments, can be by providing a user the value of the flat field correction matrix precalculated and indicating user The image of the biological sample of capture is multiplied by the value of flat field correction matrix to generate the figure through flat field correction based on pixel to pixel Picture, instruction user generate the image by Nonuniformity Correction.In some embodiments, imaging device or imaging software manufacturer Flat field correction matrix can be provided a user.In some embodiments, user method described herein and algorithm can be used to calculate flat The value of field correction matrix.
In one embodiment, a kind of to include by the method for the image of Nonuniformity Correction for generating：Calculate imaging The relative illumination of the imaging len of multiple pixels on sensor；It is inverted to relative illumination to generate flat field correction matrix.For example, Imaging device manufacturer or user produce flat field correction matrix in case using in the future.
The image through flat field correction obtained by disclosed method shows each image of the biological sample of capture Signal level accuracy, and it is unrelated with its position in imaging surface, imaging region or visual field.Figure through flat field correction As for by the image of Nonuniformity Correction.
The example of flat field correction calibration
The image device of the disclosure or a kind of exemplary application pattern of imaging system are used as under chemiluminescence pattern To the image device that biomolecule (protein and nucleic acid such as, but not limited in gel or trace) is imaged, sent out in chemistry Under optical mode, chemiluminescence sample shines without exterior lighting and excitation.As described above, chemiluminescence is carried out Suffered from a problem that when imaging be image heterogeneity, wherein when chemiluminescence sample be placed in imaging surface (such as, scheme Component 110, imaging region, visual field, sample screen or sample tray in 1) different location when, even for identical sample (same protein in such as gel or trace or nucleic acid bands), picture signal is also different.
The problem is shown by using photometer with reference to microplate.In an example, Figure 15 shows photometer reference plate 1500 perspective view.However, it is known in the art that any photometer reference plate be used equally for that the problem is shown.Photometer refers to Plate 1500 has one or more radiation spots.Eight radiation spots shown on photometer 1500 are numbered as 1501-1508. Radiation is stopped (for example, 1501-1507) by seven in radiation spot.Only one (for example, most bright) hot spot 1508 at Picture is at the different location of imaging surface (on the diagonal line of imaging screen).The hot spot 1508 of photometer reference plate 1500 is set to At different location in visual field (or sample screen).Constant time for exposure acquisition pair is used in the various pieces of imaging surface The image of the hot spot 1508 of photometer 1500, and by image stack.
Figure 16 A show the image 1600 of the hot spot 1508 of photometer 1500.Figure 16 B are shown according to one embodiment from this public affairs Figure 161 0 of the signal of the hot spot 1508 acquired at each position on the imaging surface for the image device opened.The figure of Figure 16 B 1610 displays, the signal from same spot 1508 seem there is different intensity at the different location of imaging screen, i.e., Make all signal all sames, because these signals are sent out by identical launch spot 1508 on photometer 1500.Such as Figure 16 A and figure Shown in 16B, due to being imaged at the different location of imaging screen/sample screen, the identical letter from chemiluminescence sample Number seem different.This signal difference caused by position is since a kind of characteristic i.e. relative illumination of imaging len causes. In view of the heterogeneity of image, whether user can not determine imaging difference by biomolecule (protein, core in various luminous samples Acid, DNA etc.) concentration difference cause or signal difference whether by sample strip band sample screen different location be imaged Cause.Therefore, reliable and accurate quantitative information can not be obtained using current imaging method.
Above-mentioned signal difference may be in the imaging system by the disclosure caused by the relative illumination of imaging len.It contrasts Degree is to indicate a kind of mode of halation and the comprehensive effect to roll-off in imaging len, and lead on sensors at any point Illumination percentage indicate, be normalized to the position for having in the visual field of maximal illumination.Halation and to roll-off be to contrast Two kinds of individual compositions of degree.Since above-mentioned signal difference is caused by relative illumination (that is, one of characteristic of imaging len), because This can then correct this species diversity if it is known that relative illumination.Correction course includes the relative illumination data based on imaging len Flat field correction principal matrix file is created, flat field correction principal matrix is normalized based on the maximum value in matrix, and should Flat field correction (FF) principal matrix is applied to the image captured from system.
Figure 17 shows Figure 170 0 of the relative illumination of the imaging len of the disclosure according to one embodiment.By linear or Non-linear curve fitting returns the formula that can get the curve in Figure 170 0.For discrete data point, homing method can be applied For series of points matched curve to find out best-fit equation.Then, identified formula can be used to calculate imaging sensing Any position contrasts the number of degrees on device.In one embodiment of the disclosure, the algorithm of flat field correction includes the following steps：
● step 1- calculates the opposite of all pixels on imaging sensor based on returning identified formula from curve matching Illumination number；
● step 2- is inverted to the number in step 1, and inverse is normalized based on the maximum value in inverse To generate flat field correction principal matrix；With
● the image of the biological sample captured is multiplied by step 2 by step 3- in Image Acquisition, based on pixel to pixel Middle created matrix applies the final image of flat field correction processing to generate.
The accuracy of the signal level of all bands in sample is shown through flat field correction treated image.
Figure 18 A show the image 1800 of Figure 16 A by flat field correction processing according to one embodiment, and Figure 18 B show Go out the ratio of the spot intensity and center spot in Figure 18 A.In other words, Figure 18 B show those hot spots in flat field correction processing Front and back relative percentage.As shown, before application flat field correction, each spot intensity and the light with maximum intensity value The ratio (Figure 18 A) of spot can be about 0.7, and after flat field correction is handled, which increases to greater than 0.95.As such, answering After flat field correction, significant illuminance compensation is obtained.
Figure 19 A show that according to one embodiment, chemiluminescence sample is in visual field middle position under 1 times of scaling Image 1900, Figure 19 C show under 1 times of scaling chemiluminescence sample visual field upper right angular position image 1910, and And Figure 19 B show that chemiluminescence sample is in the image 1920 in visual field centre position and upper right angular position under 1 times of scaling.
Figure 20 A show to carry out the image 2000 of quantitative analysis according to two row band of one embodiment pair, and Figure 20 B show the Figure 20 10 of intensity of a line band before flat field correction, Figure 20 C show intensity of the first row band after flat field correction Figure 20 20, Figure 20 D show Figure 20 30 of intensity of the second row band before flat field correction, and Figure 20 E show the second row Figure 20 40 of intensity of the band after flat field correction.As shown, the intensity of single band (has mutually same with imaging surface This) on sample position and change, and after flat field correction, intensity does not change with the position on imaging surface.
Generate flat field correction (FF) principal matrix
Figure 21 A show the relative illumination and sensor image height of the imaging len when amplifying 1 times according to one embodiment Table 2100, and Figure 21 B show the relative illumination of the ccd sensor imaging len and table of picture altitude when amplifying 2 times Lattice 2110.
Figure 22 shows the curve graph relative to the symmetrical relative illumination of image sensor center according to one embodiment 2200.Maximum picture height is about 8mm.It is simulated from 0mm to 8mm.
Figure 23 A show Figure 23 00 according to one embodiment optimum fit curve when amplifying 1 times, and Figure 23 B are shown Figure 23 10 of optimum fit curve when amplifying 2 times.Curve can be primary multinomial, quadratic polynomial, cubic polynomial etc.. Picture altitude is calculated using the formula：
Wherein h indicates the height (being indicated with mm) of the center pixel of distance detection sensor, xcIndicate the x of center pixel Coordinate, and ycIndicate the y-coordinate of center pixel.Pixels tall in the example is 3.69 μm/pixel.In view of this point, When amplifying 1 times, relative illumination is calculated using the formula of optimum fit curve：
RI=-0.3654h2-3.1275h+100.15
Wherein RI indicates relative illumination (%), 0≤RI≤100.
For Bin 1x1 images：
Width=3360 pixels → xc=1690
Highly=2704 pixels → yc=1352
For pixel (1,1)
RI=-0.3654 × 7.980922-3.1275×7.98092+100.15
=51.9%
Figure 24 shows the analog image 2400 when amplifying 1 times according to one embodiment.
Figure 25 shows the flat field correction main picture 2500 according to one embodiment.The value of each pixel can in master image To be equal to RI-1。
The application of flat field correction master image
Figure 26 A show according to one embodiment amplify 1 times when the image with the sample in visual field centre position 2600, Figure 26 C show the image 2610 with the sample in visual field upper right Angle Position when amplifying 1 times, and Figure 26 B are shown Image 2620 with the sample in visual field centre position and upper right Angle Position.In this example, sample is ProBloot membrane, It is 5 that it, which has the protein with the visual equivalent of chemiluminescent substrate, position in storehouse number, and scaling can be 1 times or 2 times, gain For high (for example, 55), and the term of validity is 60 seconds.
Flat field principal matrix is applied to the image in Figure 26 A-26C.It can be taken in selected item and draw rectangular mask, and And Average pixel intensity is measured.Using the macro mensuration region for ensuring measured each band for applying flat field correction Image before principal matrix and after application flat field correction principal matrix is in identical position.Although ProBloot membrane is prepared as With equal albumen quality, but the signal value between different bands still has variation.It is same for each independent band Signal strength in row should be similar, and unrelated with film location.
Figure 27 shows the image 2700 of each eight sample strip bands of two rows according to one embodiment.
It is flat in application that Figure 28 A show that Figure 28 00 of the first row before flat field correction according to one embodiment, Figure 28 B are shown Figure 28 10 of the first row, Figure 28 C show the figure of the second row before application flat field correction principal matrix after field correction principal matrix 2820, and Figure 28 D show Figure 28 30 of the second row after application flat field correction principal matrix.Times magnification in Figure 28 A-28D Number is 1 times.Before application flat field correction principal matrix, the difference of ADU (AD conversion unit) value be by image device surface/ The position of film causes on image device screen.After application flat field correction principal matrix, ADU values are similar, unrelated with the position of film.
Figure 29 A show that Figure 29 00 according to one embodiment the first row before application flat field correction principal matrix, Figure 29 B show Go out Figure 29 10 of the first row after application flat field correction principal matrix, Figure 29 C are shown the before application flat field correction principal matrix Figure 29 20 of two rows, and Figure 29 D show Figure 29 30 of the second row after application flat field correction principal matrix.In Figure 29 A-29D Amplification factor be 2 times.
Figure 30 show according to one embodiment application flat field correction principal matrix before and after film location (for example, at Centre position, upper right Angle Position as film and its sample strip band on device surface/image device screen).As shown, answering Before flat field correction principal matrix, the position in the upper right corner is relatively fuzzy, and after applying flat field correction principal matrix, band has Similar brightness.
Although illustrating that the broad range of numberical range of the present invention and parameter are approximations, illustrated in particular instance Numerical value is report reported as precisely as possible.However, any numerical value all includes inherently certain error, surveyed in their own experiment Standard deviation present in fixed will necessarily cause this error.In addition, all ranges disclosed herein are interpreted as covering Any and all subranges included in it.For example, the range of " being less than 10 " may include between minimum value of zero and maximum value 10 Between any and all subrange, that is to say, that there is the minimum value equal to or more than zero and maximum equal to or less than 10 It is worth any and all subranges of (such as 1 to 5).In some cases, as negative value can be presented in the numerical value for parameter states. In this case, negative value, such as -1, -2, -3, -10, -20, -30 etc. can be used in the example value for being set fourth as the range of " being less than 10 ".
Although having referred to the religious doctrine of its exemplary embodiment, those skilled in the art will be to described implementation Example is carry out various modifications without departing from true spirit and range.Term as used herein and description only by means of illustrate illustrate simultaneously And it is not intended to limit.Exactly, although describing method by example, it can be different from illustrated order or hold simultaneously Row method and step.In addition, with regard to used in embodiment and claims term " include (including, includes) ", For " having (having, has, with) " or the degree of its version, such term is intended to be similar to term "comprising" Mode include.As used herein, about term " in the one or more " meaning for enumerating (A and B) of all items Individual A, individual B or A and B.Those skilled in the art will realize that these and other variation may such as exist In spirit and scope defined in following claims and its equivalent.
The considerations of from specification of the invention disclosed herein with putting into practice, other embodiments of the invention will be to affiliated neck The technical staff in domain is apparent.Wish only to be considered as specification and example illustratively, wherein the true scope of the present invention It is indicated by following claims with spirit.
As used herein, term " inside " and " outside ", "up" and "down", " upward " and " downward ", " on " and " it Under ", " inside " and " outside " and other similar terms used herein refer to the relative position of each other, it is not intended that indicate specific Direction or spatial orientation.Term " coupling ", " coupling ", " connection (connect and connection) ", " connection ", " with ... connect " and " making connection " refer to " with ... be directly connected to " or " connected by one or more intermediary elements or component It connects ".

Claims (29)

1. a kind of lighting system, including：
Surface is configured to imageable target is placed on it；
Light source is configured to project light beam；
Beam splitter, the beam splitter are configured to that the first light beam and the second light beam will be separated into from the light beam of the light source；And
First speculum, first mirror arrangement, first light beam from the beam splitter that is reflection is to provide to institute State the first light beam of the reflection that surface is illuminated.
2. lighting system according to claim 1, wherein second light beam illuminates the surface.
3. lighting system according to claim 1 further includes the second speculum, second mirror arrangement is reflection institute The second light beam is stated to provide the second light beam of the reflection illuminated to the surface.
4. lighting system according to claim 3, wherein the second light beam of the first light beam of the reflection and the reflection It provides and the off-axis on the surface is illuminated.
5. lighting system according to claim 3, wherein the second light beam of the first light beam of the reflection and the reflection Illumination to the substantial symmetry on the surface is provided.
6. lighting system according to claim 1, wherein there is the light beam light beam luminous power, first light beam to have First light beam luminous power and the second light beam have the second light beam luminous power, and the wherein described first light beam luminous power and second Light beam luminous power is respectively at least the 40% of the light beam luminous power.
7. lighting system according to claim 6, wherein the first light beam luminous power and the second light beam luminous power are respectively It is at least the 45% of the light beam luminous power.
8. lighting system according to claim 7, wherein the first light beam luminous power and the second light beam luminous power are basic It is upper equal.
9. lighting system according to claim 3 further includes third speculum, wherein the first light beam of the reflection or institute The second beam configuration for stating reflection is that the third speculum is reflected before being illuminated to the surface.
10. lighting system according to claim 1, wherein be configured to will be from the light of the light source for the beam splitter Beam is separated into first light beam, second light beam and third beam.
11. lighting system according to claim 1, wherein the beam splitter includes prism, plate, dielectric mirror, metal Speculum, beam splitter cube, fiber optic splitter or optical fiber are coated, the optic fiber configureing is to generate two or more outputs Light is set to collimate bunchy before beam.
12. lighting system according to claim 1, wherein first light beam reflects the beam splitter, and it is described Second light beam passes through the beam splitter.
13. lighting system according to claim 3 further includes the second beam splitter, second beam splitter is configured to institute The first light beam for stating reflection is separated into two reflecting bundles, and the off-axis that described two reflecting bundles provide the surface different angle shines It is bright.
14. lighting system according to claim 13 further includes third beam splitter, the third beam splitter is configured to institute The second light beam for stating reflection is separated into two reflecting bundles, and the off-axis that described two reflecting bundles provide the surface different angle shines It is bright.
15. lighting system according to claim 1, wherein the center at the center of first light beam and second light beam Between angle be about 62 ° to about 68 °.
16. lighting system according to claim 1, wherein the of the center of the first light beam of the reflection and the reflection Angle between the center of two light beams is about 106 ° to about 114 °.
17. lighting system according to claim 3, wherein from the beam splitter to first speculum to the surface The first distance be substantially equal to the second distance from the beam splitter to second speculum to the surface.
18. lighting system according to claim 1, wherein the strand-separating appts have first end and the second end, and The wherein described beam splitter is variable beamsplitter, the institute that the variable beamsplitter is got in the first end and the second end The luminous power for stating light beam is different.
19. lighting system according to claim 1 further includes imaging system, wherein the imaging system includes：
Transmitting mirror；With
Capture device is configured to set by extending, reflecting the transmitting mirror from the imageable target and reach the capture The image of imageable target described in standby trace capture, wherein the transmitting mirror, described capture device or both are each configured to relatively It is moved in the diagonal directions in the surface to shorten the length in the path.
20. lighting system according to claim 19, wherein the capture device and the transmitting mirror are both in difference It is diagonally adjacent mobile simultaneously.
21. lighting system according to claim 19, wherein the capture device includes lens, optical filter and camera, and And the wherein described optical filter is positioned between the lens and the camera.
22. lighting system according to claim 19, wherein the path is reflected from the region of the transmitting mirror, and And the wherein described region is moved in the capture device, the transmitting mirror or both both with respect to the surface in the diagonal directions It is reduced when dynamic to shorten the length in the path.
23. lighting system according to claim 19, wherein the center in the path is reflected from the point of the transmitting mirror Go out, and the wherein described point in the capture device, the transmitting mirror or both both with respect to the surface in diagonal Upper mobile time shift is dynamic to shorten the length in the path.
24. lighting system according to claim 19, further includes：
Speculum axis, wherein the transmitting mirror is configured to diagonally adjacent move first along the speculum axis；
With
Capture device axis, wherein the capture device is configured to diagonally adjacent move second along the capture device axis.
25. lighting system according to claim 19 further includes drive block, the drive block transmits the transmitting mirror and institute The movement between capture device is stated, to make the transmitting mirror and the capture device while movement.
26. lighting system according to claim 25, further includes：
First transmission shaft, first transmission shaft extend between the capture device and the drive block, wherein the capture Equipment, described drive block or both are each configured to move along first transmission shaft；With
Second driving shaft, the second driving shaft extend between the transmitting mirror and the drive block, wherein the transmitting mirror, Described drive block or both is each configured to move along the second driving shaft.
27. lighting system according to claim 26, further includes：
Motor；With
Drive screw, the drive screw are couple to the motor, wherein the motor is configured to rotate the drive screw, and And the wherein described capture device, described speculum or both are in the diagonally adjacent movement in response to the transmission spiral shell The rotation of bar.
28. lighting system according to claim 19, further includes：
First motor is configured to make the transmitting mirror in the first diagonally adjacent movement；With
Second motor is configured to make the capture device in the second diagonally adjacent movement, wherein the speculum and described Capture device is mobile simultaneously, and the wherein described speculum and the capture device are moved with fixed rate relative to each other.
29. lighting system according to claim 19 further includes second light source, the second light source is configured to catch from described Outgoing beam below equipment is obtained, wherein the lower end of the transmitting mirror is positioned in the lower end of the capture device, or even when described The length in path is also such when minimizing so that the lower end of the speculum does not stop the light beam.
CN201721329355.0U 2016-10-13 2017-10-13 Lighting system Active CN207908817U (en)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US201662408018P 2016-10-13 2016-10-13
US62/408,018 2016-10-13
US201762508747P 2017-05-19 2017-05-19
US62/508,747 2017-05-19

Publications (1)

Publication Number Publication Date
CN207908817U true CN207908817U (en) 2018-09-25

Family

ID=61935380

Family Applications (2)

Application Number Title Priority Date Filing Date
CN201710960925.4A Pending CN107942522A (en) 2016-10-13 2017-10-13 For object is illuminated and is imaged equipment, System and method for
CN201721329355.0U Active CN207908817U (en) 2016-10-13 2017-10-13 Lighting system

Family Applications Before (1)

Application Number Title Priority Date Filing Date
CN201710960925.4A Pending CN107942522A (en) 2016-10-13 2017-10-13 For object is illuminated and is imaged equipment, System and method for

Country Status (1)

Country Link
CN (2) CN107942522A (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113570604A (en) * 2021-09-28 2021-10-29 武汉精创电子技术有限公司 Automatic generation method and device for crystal grain detection sample

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2009121068A2 (en) * 2008-03-28 2009-10-01 Contrast Optical Design & Engineering, Inc. Whole beam image splitting system
US8502867B2 (en) * 2010-03-19 2013-08-06 Lightspeed Genomics, Inc. Synthetic aperture optics imaging method using minimum selective excitation patterns
IL243163B (en) * 2014-12-18 2021-02-28 Gordon Noam A lighting method for imaging specular reflections from a wide range of angles in a visual inspection system

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113570604A (en) * 2021-09-28 2021-10-29 武汉精创电子技术有限公司 Automatic generation method and device for crystal grain detection sample

Also Published As

Publication number Publication date
CN107942522A (en) 2018-04-20

Similar Documents

Publication Publication Date Title
US20230273448A1 (en) Systems and methods for illuminating and imaging objects
US8842173B2 (en) Biological image acquisition device
JP7424286B2 (en) Fluorescence observation device and fluorescence observation method
US7864369B2 (en) Large-area imaging by concatenation with array microscope
US9632301B2 (en) Slide scanner with a tilted image
US9395527B2 (en) Fluorescent microscope for observing multiple fluorescent images, fluorescent image surveying method using the same, and multiple fluorescent image observing system
JP2009532732A (en) Confocal microscope having a two-dimensional array of light emitting diodes
US11269171B2 (en) Spectrally-resolved scanning microscope
CN110114709A (en) Determine the method and microscope of fluorescence intensity
CN110140129A (en) It is imaged using the imaging of the low resolution slide glass of double optics path and single imaging sensor and slide labels imaging and high-resolution slide glass
CN207908817U (en) Lighting system
WO2021177446A1 (en) Signal acquisition apparatus, signal acquisition system, and signal acquisition method
US20210360140A1 (en) Multimodality multiplexed illumination for optical inspection systems
WO2007135091A2 (en) Module for reading from a biochip
WO2022138374A1 (en) Data generation method, fluorescence observation system, and information processing device
CN114460020B (en) Hyperspectral scanning system and method based on digital micro-reflector
US20110013008A1 (en) Device for acquiring image of living body
US10627638B2 (en) Devices, systems, and methods for illuminating objects
CN109758096A (en) A kind of survey light feedback fluorescence endoscopy system and fluorescence image processing method
JP2021139927A (en) High-speed photographing device
JP4983929B2 (en) Biological image acquisition device

Legal Events

Date Code Title Description
GR01 Patent grant
GR01 Patent grant