Summary of the invention
In consideration of it, it is necessary to provide a kind of image definition analysis method, Atomatic focusing method, gene sequencers, gene
Sequencing system and storage medium can optimize the operation for judging fluorescent image clarity.
On the one hand the embodiment of the present invention provides a kind of image definition analysis method, the described method comprises the following steps:
An at least fluorescent image for biochip is obtained, is provided with multiple sites (spot) on the biochip, it is described
Site is used to combine nucleic acid molecules, and the nucleic acid molecules carry fluorophor;
The fluorescent image is subjected to discrete Fourier transform;
Fourier's amplitude figure of the fluorescent image after acquisition discrete Fourier transform;
The corresponding Fourier spectrum of the fluorescent image is obtained according to Fourier's amplitude figure;
The Fourier spectrum is obtained in an at least peak value in high frequency region;
The Fourier spectrum is obtained in the mean value of low-frequency region;
Calculate the difference degree of the peak value Yu the mean value;
When the difference degree is greater than a preset threshold, determine that the fluorescent image is clear.
Further, described by the fluorogram in above-mentioned image definition analysis method provided in an embodiment of the present invention
Include: as carrying out discrete Fourier transform
Obtain number of pixels of the fluorescent image in a first direction and in second direction, the first direction perpendicular to
The second direction;
Obtain the original two dimensional image of the fluorescent image;
The fluorescent image is subjected to discrete Fourier transform using the number of pixels and the original two dimensional image.
Further, described according in Fu in above-mentioned image definition analysis method provided in an embodiment of the present invention
Leaf amplitude figure obtains the corresponding Fourier spectrum of the fluorescent image
Fourier's amplitude figure is transformed into polar coordinate system from cartesian coordinate system;
The Fourier spectrum is obtained by carrying out Integral Processing to the angle in the polar coordinate system.
Further, in above-mentioned image definition analysis method provided in an embodiment of the present invention, the peak value is selected from described
Local maximum value set of the Fourier spectrum in high frequency region.
Further, in above-mentioned image definition analysis method provided in an embodiment of the present invention, the peak value is Fu
In leaf compose maximum value and second largest value in the local maximum value set in high frequency region.
Further, described to calculate the peak value in above-mentioned image definition analysis method provided in an embodiment of the present invention
Include: with the step of difference degree of the mean value
Calculate the sum of the maximum value and described second largest value in local maximum value set described in the Fourier spectrum with
The ratio of the mean value.
Another further aspect of the embodiment of the present invention also provides a kind of Atomatic focusing method, and the Atomatic focusing method includes:
At least fluorescent image for being located at the biochip of first position is obtained, is provided with multiple positions on the biochip
Point, the site are used to combine nucleic acid molecules, and the nucleic acid molecules carry fluorophor;
Judge whether the fluorescent image is clear using image definition analysis method described in any of the above embodiments;
If judging result is no, fluorescent image of the acquisition positioned at the biochip of the second position.
Another further aspect of the embodiment of the present invention also provides a kind of gene sequencing system, and the gene sequencing system includes:
Image collection module is provided with more for obtaining an at least fluorescent image for biochip on the biochip
A site, the site are used to combine nucleic acid molecules, and the nucleic acid molecules carry fluorophor;
Discrete Fourier transform module, for the fluorescent image to be carried out discrete Fourier transform;
Fourier's amplitude obtains module, for obtaining Fourier's amplitude of the fluorescent image after discrete Fourier transform
Figure;
Fourier spectrum obtains module, for obtaining the corresponding Fourier of the fluorescent image according to Fourier's amplitude figure
Spectrum;
Peak value obtains module, for obtaining the Fourier spectrum in an at least peak value in high frequency region;
Mean value obtains module, for obtaining the Fourier spectrum in the mean value of low-frequency region;
Difference degree computing module, for calculating the difference degree of the peak value Yu the mean value;
Judgment module, for determining that the fluorescent image is clear when the difference degree is greater than a preset threshold.
Another further aspect of the embodiment of the present invention also provides a kind of gene sequencer, and the gene sequencer includes processor, institute
Processor is stated for realizing image definition described in above-mentioned any one point when executing the computer program stored in memory
The step of analysis method.
Another further aspect of the embodiment of the present invention also provides a kind of non-volatile computer readable storage medium storing program for executing, is stored thereon with meter
Calculation machine program, the computer program realize image definition analysis method described in above-mentioned any one when being executed by processor
The step of.
Image definition analysis provided in an embodiment of the present invention and focusing method, gene sequencer and system and storage are situated between
Matter.The fluorescent image is carried out discrete Fourier transform by at least fluorescent image for obtaining biochip;Obtain direct computation of DFT
Fourier's amplitude figure of the fluorescent image after leaf transformation;It is corresponding that the fluorescent image is obtained according to Fourier's amplitude figure
Fourier spectrum;The Fourier spectrum is obtained in an at least peak value in high frequency region;The Fourier spectrum is obtained in low frequency
The mean value in region;Calculate the difference degree of the peak value Yu the mean value;When the difference degree is greater than a preset threshold, sentence
The fixed fluorescent image is clear.Sample frequency (the i.e. pixel of single DNA nanosphere of the embodiment of the present invention for DNA nanosphere
Number) it is insensitive, the sample frequency of DNA nanosphere, which changes, will not change the size of peak value, thus be implemented using the present invention
Example, the gene sequencer of biochip and different model for different model, it is not necessary to modify evaluation functions;The present invention is implemented
Example, can calculate the image definition of entire fluorescent image or localized fluorescence image, and it is clear not need to evaluate using a single point
It spends, thus is not easily susceptible to the influence of noise spot using the embodiment of the present invention;The embodiment of the present invention carries out Fourier's amplitude
It is calculated again after normalized, and considers the intensity of low frequency signal, thus light can be reduced using the embodiment of the present invention
According to the influence generated with background value transformation.
Specific embodiment
In order to be more clearly understood that the above objects, features, and advantages of the embodiment of the present invention, with reference to the accompanying drawing and
The present invention will be described in detail for specific embodiment.It should be noted that in the absence of conflict, the embodiment party of the application
Feature in formula can be combined with each other.
Embodiment in the following description, numerous specific details are set forth in order to facilitate a full understanding of the present invention, described reality
The mode of applying is some embodiments of the invention, rather than whole embodiments.Based on the embodiment in the present invention, ability
Domain those of ordinary skill every other embodiment obtained without making creative work, belongs to the present invention
The range of embodiment protection.
Unless otherwise defined, all technical and scientific terms used herein and the technology for belonging to the embodiment of the present invention
The normally understood meaning of the technical staff in field is identical.Term as used herein in the specification of the present invention is intended merely to
The purpose of specific embodiment is described, it is not intended that in the limitation embodiment of the present invention.
Fig. 1 is the flow chart of image definition analysis method provided in an embodiment of the present invention.As shown in Figure 1, described image
Clarity analysis method may include steps of:
S101: an at least fluorescent image for biochip is obtained, multiple sites, institute's rheme are provided on the biochip
For point for combining nucleic acid molecules, the nucleic acid molecules carry fluorophor.
In present embodiment, the biochip can be gene sequencing chip, and the fluorescent image, which can be, to be sequenced
Fluorophor described in journey shines obtained image, can use micro- camera continuously take pictures to biochip it is several to obtain
Fluorescent image.It should be noted that herein " fluorescent image " refer to micro- camera once shoot a resulting visual field (FOV,
Field of view) image again through partial enlargement image obtained.The visual field of micro- camera is smaller, about 768.6 μm of *
648μm.In sequencing procedure, several hundred width fluorescent images can be shot to biochip.The horizontal direction in each visual field with it is vertical
Several trajectory lines are respectively distributed on direction, the intersection point of trajectory line (trackline) is track cross point (trackcross).Each
Region between adjacent two horizontal directions and the trajectory line of vertical direction in the visual field is known as a block (block), Mei Gequ
Site (spot) it has been covered in block, it is preferable that the site is uniformly distributed, and the site is used to combine nucleic acid molecules, the core
Acid molecule carries fluorophor.
S102: the fluorescent image is subjected to discrete Fourier transform.
It may include: to obtain the fluorogram by fluorescent image progress discrete Fourier transform in present embodiment
As the number of pixels in a first direction and in second direction, the first direction is perpendicular to the second direction.Described first
Direction can be horizontal direction, and the second direction can be vertical direction.The original two dimensional image of the fluorescent image is obtained,
The original two dimensional image can be clearly fluorescent image, will using the number of pixels and the original two dimensional image
The fluorescent image carries out discrete Fourier transform.
In one embodiment, the fluorescent image can be saved as to tif format, the fluorescent image of the tif format
In include the fluorescent image file header, the file header can carry the number of pixels in first direction and second direction,
By parsing the file header, number of pixels of the available fluorescent image in a first direction and in second direction.
S103: Fourier's amplitude figure of the fluorescent image after discrete Fourier transform is obtained.
In present embodiment, after the fluorescent image is carried out discrete Fourier transform, the fluorescence can be obtained
Fourier's amplitude figure of image.In Fourier's amplitude figure the gray value of each pixel represent respective frequencies amplitude it is big
It is small.
S104: the corresponding Fourier spectrum of the fluorescent image is obtained according to Fourier's amplitude figure.
In present embodiment, it is described the corresponding Fourier spectrum of the fluorescent image is obtained according to Fourier's amplitude figure can
To include the following steps: Fourier's amplitude figure transforming to polar coordinate system from cartesian coordinate system, by being sat to the pole
Angle in mark system carries out Integral Processing and obtains the Fourier spectrum.The cartesian coordinate system is that rectangular coordinate system and oblique angle are sat
The general designation of system is marked, two number axis linear modules that origin is intersected in the cartesian coordinate system are equal.The polar coordinate system is
Refer to the coordinate system being planar made of pole, polar axis and polar diameter.The cartesian coordinate system and the polar coordinate system can be with
Mutually conversion.
S105: the Fourier spectrum is obtained in an at least peak value in high frequency region.
In present embodiment, the peak value is selected from the Fourier spectrum in the local maximum value set in high frequency region.
The peak value can be maximum value and second largest value of the Fourier spectrum in the local maximum value set in high frequency region, institute
Stating peak value can also be the preceding n in the local maximum value set in high frequency region biggish values of the Fourier spectrum,
In, n is more than or equal to 1.The high frequency area size and the original two dimensional image size of the fluorescent image of acquisition have
It closes.It is described by taking the original two dimensional image of the fluorescent image that size is 640 pixel *, 540 pixel as an example in present embodiment
High frequency region may include 150Hz-300Hz.It is understood that 150Hz expression is every 2.79 ((1/150) poles *
Coordinate system maximum radius) a pixel size, there is a sampled signal.The polar coordinate system maximum radius is 419 (sqrt
(320*320+270*270)) pixel.
S106: the Fourier spectrum is obtained in the mean value of low-frequency region.
In present embodiment, by taking the original two dimensional image of the fluorescent image that size is 640 pixel *, 540 pixel as an example,
The low-frequency region may include 30Hz-150Hz.The Fourier spectrum may include Fourier in the mean value of low-frequency region
Compose the average value in low-frequency region Fourier's amplitude.
S107: the difference degree of the peak value Yu the mean value is calculated.
In present embodiment, the difference degree may include maximum value described in the Fourier spectrum and the second largest value
Carry out the ratio of sum and the mean value.It is understood that the ratio is bigger if fluorescent image is more clear;If
Fluorescent image is fuzzyyer, then the ratio is smaller.
S108: when the difference degree is greater than a preset threshold, determine that the fluorescent image is clear.
In present embodiment, it is pre-set based on experience value that the preset threshold can be those skilled in the art.It can
With understanding, the difference degree is bigger, then the fluorescent image is more clear;The difference degree is smaller, then the fluorescence
Image is fuzzyyer.If the difference degree is less than preset threshold, it can control motor mobile platform and obtain positioned at the second position
Biochip fluorescent image.The second position refers to position of the biochip with respect to object lens.
Image definition analysis method provided in an embodiment of the present invention obtains an at least fluorescent image for biochip, will
The fluorescent image carries out discrete Fourier transform;Fourier's amplitude of the fluorescent image after acquisition discrete Fourier transform
Figure;The corresponding Fourier spectrum of the fluorescent image is obtained according to Fourier's amplitude figure;The Fourier spectrum is obtained in middle height
An at least peak value for frequency field;The Fourier spectrum is obtained in the mean value of low-frequency region;Calculate the peak value and it is described
The difference degree of value;When the difference degree is greater than a preset threshold, determine that the fluorescent image is clear.It is real using the present invention
Example is applied, the operation for judging fluorescent image clarity can be optimized.
Fig. 2 is the flow chart of Atomatic focusing method provided in an embodiment of the present invention.As shown in Fig. 2, the auto-focusing side
Method may include steps of:
S201: at least fluorescent image for being located at the biochip of first position is obtained, is provided on the biochip
Multiple sites, it is preferable that the site is uniformly distributed, and the site is used to combine nucleic acid molecules, and the nucleic acid molecules carry glimmering
Light group.
S202: judge whether the fluorescent image is clear, when the judgment result is no, enter step 203;Work as judging result
When to be, terminate process.
In present embodiment, the difference degree of the peak value Yu the mean value can be calculated, by judging the difference journey
Whether degree is greater than the method for preset threshold, judges whether the fluorescent image is clear.It is understood that if the difference degree
Greater than preset threshold, then determine that the fluorescent image is clear.
S203: the fluorescent image for being located at the biochip of the second position is obtained.
In present embodiment, the platform for loading biochip can be moved to second by controlling motor mobile platform
Position obtains the fluorescent image for being located at the biochip of the second position.It is understood that being located at the second position in described obtain
Biochip fluorescent image after, can continue judgement be located at the second position biochip fluorescent image it is whether clear
It is clear, when the judgment result is no, obtain the fluorescent image for being located at the biochip of the third place.It is provided in an embodiment of the present invention from
Dynamic focusing method, obtains at least fluorescent image for being located at the biochip of first position, judges whether the fluorescent image is clear
It is clear, if the fluorescent image is unintelligible, obtain the fluorescent image for being located at the biochip of the second position.Implemented using the present invention
Example, the gene sequencer of biochip and different model for different model, it is not necessary to modify evaluation function, flexibility is higher;
The embodiment of the present invention does not need to evaluate clarity using a single point, thus is not easily susceptible to the influence of noise spot, and the present invention is implemented
Example calculates again after Fourier's amplitude is normalized, and considers the intensity of low frequency signal, thus benefit
The influence that illumination and background value transformation generate can be reduced with the embodiment of the present invention.
It is the detailed description for being provided for the embodiments of the invention method progress above.The embodiment of the present invention is mentioned below
The gene sequencer of confession is described.
The embodiment of the present invention also provides a kind of gene sequencer, including memory, processor and storage are on a memory simultaneously
The computer program that can be run on a processor, the processor realize institute in any of the above-described embodiment when executing described program
The step of image definition analysis method stated.It should be noted that the gene sequencer may include chip platform, optics
System, liquid channel system.Wherein, the chip platform can be used for loading biochip, and the optical system can be used for obtaining
Fluorescent image, the liquid channel system can be used for carrying out biochemical reaction using preset reagent.
Fig. 3 is the structural schematic diagram of the gene sequencer of an embodiment of the present invention.As shown in figure 3, gene sequencer 1 wraps
Memory 10 is included, gene sequencing system 100 is stored in memory 10.The available biological core of gene sequencing system 100
The fluorescent image is carried out discrete Fourier transform by an at least fluorescent image for piece;Described in obtaining after discrete Fourier transform
Fourier's amplitude figure of fluorescent image;The corresponding Fourier spectrum of the fluorescent image is obtained according to Fourier's amplitude figure;It obtains
Take the Fourier spectrum in an at least peak value in high frequency region;The Fourier spectrum is obtained in the mean value of low-frequency region;
Calculate the difference degree of the peak value Yu the mean value;When the difference degree is greater than a preset threshold, the fluorescence is determined
Image clearly.Using the embodiment of the present invention, the operation for judging fluorescent image clarity can be optimized.
The gene sequencing system 100 can also obtain an at least fluorogram for the biochip positioned at first position
Picture judges whether the fluorescent image is clear, if the fluorescent image is unintelligible, obtains the biochip for being located at the second position
Fluorescent image.Using the embodiment of the present invention, the gene sequencer of biochip and different model for different model is not necessarily to
Evaluation function is modified, flexibility is higher;The embodiment of the present invention does not need to evaluate clarity using a single point, thus is not easily susceptible to make an uproar
The influence of sound point, and Fourier's amplitude is normalized and calculates again later by the embodiment of the present invention, and considers
Low frequency signal it is slight, thus the influence that illumination and background value transformation generate can be reduced using the embodiment of the present invention.
In present embodiment, gene sequencer 1 can also include display screen 20 and processor 30.Memory 10, display screen
20 can be electrically connected with processor 30 respectively.
The memory 10 can be different type storage equipment, for storing Various types of data.For example, it may be gene
Memory, the memory of sequenator 1, can also be the storage card that can be external in the gene sequencer 1, such as flash memory, SM card (Smart
Media Card, smart media card), SD card (Secure Digital Card, safe digital card) etc..In addition, memory 10 can
It can also include nonvolatile memory, such as hard disk, memory, plug-in type hard disk, intelligence to include high-speed random access memory
Energy storage card (Smart Media Card, SMC), secure digital (Secure Digital, SD) card, flash card (Flash
Card), at least one disk memory, flush memory device or other volatile solid-state parts.Memory 10 is for storing
Various types of data, for example, the types of applications program (Applications) installed in the gene sequencer 1, using above-mentioned image
Clarity analysis method and be arranged, the information such as the data that obtain.
Display screen 20 is installed on gene sequencer 1, for showing information.
Processor 30 be used to execute installed in described image clarity analysis method and the gene sequencer 1 it is all kinds of
Software, such as operating system and application display software etc..Processor 30 is including but not limited to processor (Central
Processing Unit, CPU), micro-control unit (Micro Controller Unit, MCU) etc. refers to for interpretive machine
Enable and handle the device of the data in computer software.
The gene sequencing system 100 may include one or more module, and one or more of modules are deposited
Storage in the memory 10 of gene sequencer 1 and be configured to by one or more processors (present embodiment be one processing
Device 30) it executes, to complete the embodiment of the present invention.For example, as shown in fig.4, the gene sequencing system 100 may include image
It obtains module 11, discrete Fourier transform module 12, Fourier's amplitude and obtains module 13, Fourier spectrum acquisition module 14, peak value
Obtain module 15, mean value obtains module 16, difference degree computing module 17, judgment module 18.The so-called mould of the embodiment of the present invention
Block can be the program segment for completing a specific function, the implementation procedure than program more suitable for description software in the processor.
It is understood that each embodiment in corresponding above-mentioned image definition analysis method, gene sequencing system 100
It may include part or all in each functional module shown in Fig. 4, the function of each module will introduced in detail below.It needs
It is noted that identical noun related terms and its specific solution in each embodiment of images above clarity analysis method
It releases explanation and is readily applicable to the following function introduction to each module.For the sake of saving space and avoiding repetition, herein just no longer
It repeats.
Image collection module 11 can be used for obtaining an at least fluorescent image for biochip, be provided on the biochip
Multiple sites, it is preferable that the site is uniformly distributed, and the site is used to combine nucleic acid molecules, and the nucleic acid molecules carry glimmering
Light group.
Image collection module 11 can also be used in at least fluorescent image for obtaining the biochip for being located at first position, described
Multiple sites are provided on biochip, it is preferable that the site is uniformly distributed, and the site is used to combine nucleic acid molecules, institute
It states nucleic acid molecules and carries fluorophor.
Image collection module 11 can also be used in the fluorescent image for obtaining the biochip for being located at the second position.
Discrete Fourier transform module 12 can be used for the fluorescent image carrying out discrete Fourier transform.
Fourier's amplitude, which obtains module 13, can be used for obtaining Fourier's width of the fluorescent image after discrete Fourier transform
Value figure.
Fourier spectrum, which obtains module 14, can be used for obtaining corresponding Fu of the fluorescent image according to Fourier's amplitude figure
In leaf compose.
Peak value, which obtains module 15, can be used for obtaining the Fourier spectrum in an at least peak value in high frequency region.
Mean value, which obtains module 16, can be used for obtaining the Fourier spectrum in the mean value of low-frequency region.
Difference degree computing module 17 can be used for calculating the difference degree of the peak value Yu the mean value.
Judgment module 18 can be used for determining that the fluorescent image is clear when the difference degree is greater than a preset threshold.
The embodiment of the present invention also provides a kind of non-volatile computer readable storage medium storing program for executing, is stored thereon with computer journey
Sequence, the computer program realize the step of the image definition analysis method in any of the above-described embodiment when being executed by processor
Suddenly.
If the integrated module/unit of the gene sequencing system/gene sequencer/computer equipment is with software function list
Member form realize and when sold or used as an independent product, can store in a computer-readable storage medium
In.Based on this understanding, the present invention realizes all or part of the process in above embodiment method, can also pass through calculating
Machine program is completed to instruct relevant hardware, and the computer program can be stored in a computer readable storage medium,
The computer program is when being executed by processor, it can be achieved that the step of above-mentioned each embodiment of the method.Wherein, the computer journey
Sequence includes computer program code, and the computer program code can be source code form, object identification code form, executable text
Part or certain intermediate forms etc..The computer readable storage medium may include: that can carry the computer program code
Any entity or device, recording medium, USB flash disk, mobile hard disk, magnetic disk, CD, computer storage, read-only memory (ROM,
Read-Only Memory), random access memory (RAM, Random Access Memory), electric carrier signal, telecommunications letter
Number and software distribution medium etc..
Alleged processor can be central processing unit (Central Processing Unit, CPU), can also be it
His general processor, digital signal processor (Digital Signal Processor, DSP), specific integrated circuit
(Application Specific Integrated Circuit, ASIC), ready-made programmable gate array (Field-
Programmable Gate Array, FPGA) either other programmable logic device, discrete gate or transistor logic,
Discrete hardware components etc..General processor can be microprocessor or the processor is also possible to any conventional processor
Deng the processor is the control centre of the gene sequencing system/gene sequencer, whole using various interfaces and connection
The various pieces of a gene sequencing system/gene sequencer.
The memory is deposited for storing the computer program and/or module, the processor by operation or execution
Computer program in the memory and/or module are stored up, and calls the data being stored in memory, realizes the base
Because of the various functions of sequencing system/gene sequencer.The memory can mainly include storing program area and storage data area,
In, storing program area can application program needed for storage program area, at least one function (such as sound-playing function, image
Playing function etc.) etc.;Storage data area, which can be stored, uses created data (such as audio data, phone directory according to mobile phone
Deng) etc..In addition, memory may include high-speed random access memory, it can also include nonvolatile memory, such as firmly
Disk, memory, plug-in type hard disk, intelligent memory card (Smart Media Card, SMC), secure digital (Secure Digital,
SD) block, flash card (Flash Card), at least one disk memory, flush memory device or other volatile solid-states
Part.
Fig. 5 A is please referred to, Fig. 5 A is the site distributed image on biochip provided in an embodiment of the present invention.The biology
Multiple sites are distributed on chip, it is preferred that the site is uniformly distributed.As shown in Figure 5A, biology provided in an embodiment of the present invention
Site on chip is to be uniformly distributed.The size in each site can be 900nm*900nm, and the site is received for loading DNA
Rice ball, the DNA nanosphere belong to a kind of nucleic acid molecules.Large circle point shown in Fig. 5 A indicates site, the picture of the site image
Plain size can be 300nm*300nm, it is to be understood that the DNA nanosphere loaded on each site occupies 3*3 pixel
Region.
Fig. 5 B is please referred to, Fig. 5 B is part DNA nanosphere fluorescence imaging figure on biochip provided in an embodiment of the present invention.
Dot shown in Fig. 5 B indicates the imaging of DNA nanosphere, it will be understood by those skilled in the art that in sequencing procedure, not
It is so can all combine DNA nanometers of sphere molecules on site, and be not each DNA nanometers of sphere molecule in the Shi Doufa that takes pictures every time
Light, the base fluorophor for only corresponding to current channel shines, thus DNA nanosphere shown in Fig. 5 B is luminous DNA nanometer
Ball.The spatial frequency of the DNA nanosphere is fixed, and the spatial distribution of the DNA nanosphere has periodically.It is described
DNA nanosphere can be formed by stacking by less cosine wave, thus the corresponding two dimensional discrete Fourier transform of the fluorescent image
Fourier's amplitude can occur peak value at corresponding frequencies.
Fig. 6 A is please referred to, Fig. 6 A is the regional area figure of clear fluorescent image provided in an embodiment of the present invention.The present embodiment
In, it can indicate that clearly two-dimensional fluoroscopic image, M indicate that the number of pixels in the direction x, N indicate the pixel number in the direction y with L (x, y)
Mesh, then the two-dimentional DFT (discrete Fourier transform) of the fluorescent image is expressed as follows with formula:
The corresponding inverse discrete Fourier transform of the clearly two-dimensional fluoroscopic image is expressed as follows with formula:
Fig. 6 B is please referred to, Fig. 6 B is after the regional area figure of Fig. 6 A clear fluorescent image provided is carried out Fourier transformation
Fourier's amplitude figure.As shown in Figure 6B, the clearly regional area of two-dimensional fluoroscopic image corresponding discrete Fourier transform
Fourier's amplitude is larger in some regions, and black circle is artificially added up, and shows that Fourier's amplitude at this is larger, Fu
In leaf amplitude size indicated with the gray scale of the pixel of Fourier's amplitude figure.
Fig. 7 A is please referred to, Fig. 7 A is the regional area figure of out-of-focus image provided in an embodiment of the present invention.In present embodiment,
Indicate that the two-dimensional fluoroscopic image of defocus, L (x, y) indicate that clearly two-dimensional fluoroscopic image, n indicate that noise, f indicate point with I (x, y)
Spread function, point spread function f can be approximate with two-dimensional Gaussian function.Clearly two-dimensional fluoroscopic image is carried out using formula 3 low
Pass filter:
Fig. 7 B is the Fourier's amplitude figure carried out the regional area figure of Fig. 7 A out-of-focus image provided after Fourier transformation.
As shown in Figure 7 B, the corresponding Fourier transformation of out-of-focus image is smaller in medium-high frequency region Fourier's amplitude.
Fig. 8 A, 8B are please referred to, Fig. 8 A is the corresponding Fourier of two-dimensional fluoroscopic image of defocus provided in an embodiment of the present invention
Spectrogram, Fig. 8 B are the corresponding Fourier's spectrogram of clearly two-dimensional fluoroscopic image provided in an embodiment of the present invention.Such as Fig. 8 A, 8B institute
Show, in the present embodiment, Fourier's amplitude figure is transformed into polar coordinate system from cartesian coordinate system, by the polar coordinates
Angle in system carries out Integral Processing and obtains the Fourier spectrum.The center of Fig. 8 A, 8B is arranged in the origin of the polar coordinate system.
As shown in Fig. 8 A, 8B, horizontal axis indicates frequency, and the longitudinal axis indicates the ratio of the corresponding total Fourier's amplitude of Fourier's amplitude Zhan of frequency
(i.e. normalized Fourier's amplitude).Region of interest area image with the fluorescent image of 640 pixel *, 540 pixel size is
Example, the high frequency region of the Fourier spectrum may include 150Hz-300Hz, and low-frequency region may include 30Hz-
150Hz.As shown in Figure 8 A, the corresponding Fourier's spectrogram of the two-dimensional fluoroscopic image of defocus does not occur peak in high frequency region
Value.As shown in Figure 8 B, clearly in high frequency region there is peak value, the peak in the corresponding Fourier's spectrogram of two-dimensional fluoroscopic image
Local maximum value set of the value selected from the Fourier spectrum in high frequency region indicates local maximum with { localPeaks }
Set, the peak value can be maximum value of the Fourier spectrum in the local maximum value set in high frequency region and time big
Value.The Fourier spectrum is obtained in the mean value of low-frequency region, indicates the low-frequency region with avg (lowFreq_mag)
Mean value, and calculate the ratio of maximum value described in the Fourier spectrum and the second largest value progress sum and the mean value.
The ratio is indicated with following formula:
Formula 4 is known as empty burnt evaluation function.The ratio is bigger, then fluorescent image is more clear;The ratio is smaller, then glimmering
Light image is fuzzyyer.
It is understood that the peak value can also include local maximum of the Fourier spectrum in high frequency region
Preceding n biggish values in set (n is more than or equal to 1).
In several specific embodiments provided by the present invention, it should be understood that disclosed terminal and method, it can be with
It realizes by another way.For example, system embodiment described above is only schematical, for example, the module
Division, only a kind of logical function partition, there may be another division manner in actual implementation.
It is obvious to a person skilled in the art that the embodiment of the present invention is not limited to the details of above-mentioned exemplary embodiment,
And without departing substantially from the spirit or essential attributes of the embodiment of the present invention, this hair can be realized in other specific forms
Bright embodiment.Therefore, in all respects, the present embodiments are to be considered as illustrative and not restrictive, this
The range of inventive embodiments is indicated by the appended claims rather than the foregoing description, it is intended that being equal for claim will be fallen in
All changes in the meaning and scope of important document are included in the embodiment of the present invention.It should not be by any attached drawing mark in claim
Note is construed as limiting the claims involved.Multiple units, module or the device stated in system, device or terminal claim
It can also be implemented through software or hardware by the same unit, module or device.
Embodiment of above is only to illustrate the technical solution of the embodiment of the present invention rather than limits, although referring to above preferable
The embodiment of the present invention is described in detail in embodiment, those skilled in the art should understand that, it can be to this hair
The technical solution of bright embodiment is modified or equivalent replacement should not all be detached from the embodiment of the present invention technical solution spirit and
Range.