KR101641777B1 - Image generating apparatus and image generating method - Google Patents

Abstract

An image generating apparatus is disclosed. An apparatus for generating an image according to an exemplary embodiment of the present invention includes a light source for outputting light, a random medium for scattering the output light to have a speckle pattern, a relative medium between the light output from the light source and the random medium, A sensor for sensing a fluorescent signal emitted from the fluorescent sample and converting the fluorescent signal into an electrical signal when light having the speckle pattern is irradiated to the fluorescent sample, And an image processor for obtaining a plurality of images according to each of the varying speckle patterns using the electrical signal and processing the plurality of images to generate a fluorescence image.

Description

IMAGE GENERATING APPARATUS AND IMAGE GENERATING METHOD Document Type and Number: WIPO Patent Application WO /

The present disclosure relates to an image generating apparatus and an image generating method, and more particularly, to an image generating apparatus and an image generating apparatus for collecting a fluorescence signal from a fluorescent sample when light is irradiated on the fluorescent sample to generate a high- Generating method.

Recently, studies on high resolution image generation methods have been actively conducted. Conventional optical microscopes have limited resolvability due to the diffraction limit known as abbe's law, so that the two objects must be at least half wavelength apart. Many methods have been proposed to overcome this diffraction limit. Among them, the high resolution fluorescence imaging technique has been popular because it is easy to use for bio-imaging by improving the resolution using the characteristics of the phosphor. High resolution fluorescence imaging techniques have also been proposed.

A super-resolution optics fluctuation imaging (SOFI) method has been studied as one of high resolution fluorescence imaging techniques. The SOFI method treats the blinking phenomenon as a statistical concept.

FIGS. 1A and 1B are conceptual diagrams for explaining the SOFI method. As shown in Fig. 1A, in the SOFI method, phosphors 11 and 12 may exist at different positions of the fluorescent sample 10, respectively. Each of the phosphors 11 and 12 may blink at a specific time interval due to inherent blinking properties. The SOFI method measures the flickering phenomenon of the fluorescent sample 10 at time intervals with the images 21 to 24 at certain time intervals as shown in FIG. 1B. On the other hand, the self-flickering of the phosphor 11 and the phosphor 12 has high autocorrelation characteristics. That is, in a portion where the flickering of the phosphor 11 and the flickering signal of the phosphor 12 coexist, the crosscorrelation coefficient has a low value because the signals of the two phosphors 11 and 12 are mixed, The autocorrelation coefficient of the flicker of only the phosphor 11 or the flicker of the phosphor 12 alone may have a high value. Hereinafter, such characteristics are expressed as having autocorrelation properties.

In the SOFI method, when each of the measured images 21 to 24 is multiplied by a corresponding pixel of an adjacent image in time to perform a statistical process, the self-correlation characteristic of the flickering fluorescent material 11, Only the product of the phosphors is survived, and the corresponding point spread function (PSF) is squared so that the effect of reducing the PSF is obtained. Reducing the PSF to a smaller size is equivalent to increasing the resolution of the image. From this, the SOFI method can obtain a high-resolution fluorescence image, the fluorescence image obtained from the conventional optical microscope from the left side of FIG. 1C, and the remaining image is a high-resolution fluorescence image obtained using the SOFI method. For example, in the second figure on the left side of FIG. 1C, high-resolution fluorescence images obtained through statistical processing of second order correlation are applied. When a high order correlation is applied, As shown in the figure, theoretically, the PSF can be reduced by the coefficient of the higher order term so that a higher resolution image can be obtained.

However, since the SOFI method needs to use blinking which is an intrinsic property of the phosphor and has a characteristic of flickering at different time intervals depending on the phosphor, it is disadvantageous to use a specific phosphor satisfying it. Therefore, there is a limit to apply to a biological sample using a general phosphor.

Accordingly, there is a need to develop an image generating apparatus and an image generating method capable of generating a high-resolution image by applying the SOFI method without using the flicker of the phosphor.

The following disclosure provides an image generating apparatus and an image generating method capable of generating a high-resolution image using a SOFI method without using flicker of a phosphor, which is devised in response to a technology development request while solving the above-mentioned problems .

The following disclosure discloses a method of irradiating a fluorescent sample with light having a specific pattern that varies with time having autocorrelation properties, and generates flickering of a fluorescent signal generated from the fluorescent material of the fluorescent sample from the irradiated light. The resulting flickering of the fluorescent signal can replace the flickering of the phosphor. Accordingly, there is provided an image generating apparatus and an image generating method for generating a high-resolution image using a SOFI method and a Fourier re-weighting method without using a flicker phenomenon of a phosphor.

The apparatus for generating an image according to an exemplary embodiment includes: (1) a light source for outputting light; (2) a random medium for generating a speckle pattern when light emitted from the light source is transmitted; (3) a position drive device for changing the relative position or angle between the light emitted from the light source and the random medium in order to change the speckle pattern with time; (4) An image sensor for detecting a fluorescence signal to be generated and converting the generated fluorescence signal into an electrical signal, (5) obtaining a plurality of fluorescent images over time from the fluorescence sample using the speckle pattern changed by the position drive device, (SOFI) to produce a high resolution fluorescence image.

The image processor may reconstruct an image such as a fluorescence image measured by a general microscope using the mean of the plurality of fluorescence images.

The image processor may apply a super-resolution optics fluctuation imaging (SOFI) method to the plurality of images to produce a high-resolution fluorescence image of the fluorescent sample.

Wherein the image processor applies a Fourier reweighting method (FRW) to a high-resolution image generated by processing the plurality of fluorescence images with the statistical method (SOFI) to generate fluorescence images with a higher resolution than the high- Can be generated.

The position drive device may change the speckle pattern generated from the random medium by changing the relative position or angle between the light emitted from the light source and the random medium.

The position drive apparatus can adjust the degree of change of the speckle pattern with time to a degree that changes the relative position or angle between the light emitted from the light source and the random medium.

The spark pattern generating device according to another embodiment may include a random medium and a position drive device.

According to another embodiment of the present invention, there is provided a method of generating an image, the method comprising: outputting light such that light having a speckle pattern generated from a random medium is irradiated on the fluorescent sample; detecting a fluorescent signal generated from the fluorescent sample; Obtaining a plurality of images from the fluorescence samples using a speckle pattern generated by varying a relative position or angle between light emitted from the light source and the random medium over time, By the statistical method and the Fourier re-weighting method to generate a high-resolution fluorescence image.

According to the above-described various embodiments, a method of irradiating a sample with a specific pattern having a specific pattern that changes with time having autocorrelation properties, can be applied to a sample using a SOFI method and a Fourier re- An image generating apparatus and an image generating method for generating an image can be provided. By not using the flicker of the phosphor, it can be applied to various fluorescence samples, and its application range can be extended.

In addition, the image generating apparatus and the image generating method according to various embodiments can be implemented by slightly modifying the existing SOFI scheme, thereby providing high economic efficiency and applicability.

In addition, the image generating apparatus and the image generating method according to various embodiments can be realized at a higher speed than SOFI by adjusting the change of the speckle pattern from the outside using the position driving apparatus. Accordingly, real-time imaging that can not be implemented in the existing SOFI scheme can be realized.

FIGS. 1A and 1B are conceptual diagrams for explaining the SOFI method.
Figure 1C is a high resolution fluorescence image obtained based on the SOFI method.
2 is a block diagram of an image generating apparatus according to an embodiment.
3 is a conceptual diagram for explaining generation of a speckle pattern according to an embodiment.
4 is a conceptual diagram for explaining the movement of the random medium and the change of the speckle pattern according to an embodiment.
5 is a graph for explaining the resolution enhancement based on the SOFI Fourier reweighting method (FRW).
6 is a conceptual diagram for explaining an image generating apparatus according to an embodiment.
FIG. 7 is an illustration of a plurality of images obtained from a fluorescence signal emitted from a fluorescent sample and illuminated with a random sample by a random medium and a speckle pattern varying with time by a position drive device.
8 is a graph of experimental simulation results according to an embodiment.
Fig. 9 is a graph for explaining a correlation change between speckle patterns according to the degree of movement of the random incident position drive device. Fig.
10 is a block diagram of an apparatus for generating a speckle pattern according to an embodiment.
11 is a flow diagram of a method for generating an image for a sample according to an embodiment.

In the following, some embodiments will be described in detail with reference to the accompanying drawings. However, it is not limited or limited by these embodiments. Like reference symbols in the drawings denote like elements.

Although the terms used in the following description have selected the general terms that are widely used in the present invention while considering the functions of the present invention, they may vary depending on the intention or custom of the artisan, the emergence of new technology, and the like.

Also, in certain cases, there may be terms chosen arbitrarily by the applicant for the sake of understanding and / or convenience of explanation, and in this case the meaning of the detailed description in the corresponding description section. Therefore, the term used in the following description should be understood based on the meaning of the term, not the name of a simple term, and the contents throughout the specification.

2 is a block diagram of an image generating apparatus according to an embodiment.

2, an image generating apparatus 200 according to an embodiment includes a light source 210, a random medium 220, a position drive unit 230, a sensor 240, and an image processor 250).

The light source 210 may output light having a predetermined wavelength. The light source 210 may be implemented by, for example, a laser oscillation apparatus, and there is no limitation on the type of the laser oscillation apparatus.

The light source 210 may illuminate the light towards the random medium 220. The relative position between the light source 210 and the random medium 220 may be adjusted by the position driver 230 such that light from the light source 210 is incident on the first point of the random medium 220. Meanwhile, between the light source 210 and the random medium 220, another optical system may be included to transmit light.

The random medium 220 may be a disordered matter that causes a speckle pattern to be generated. The random medium 220 is not limited as long as it can scatter the incident light so that the ray of incident light propagates through various paths so that the transmitted light has a speckle pattern.

The position drive 230 may change the position or angle of the random medium 220 to change the relative position or angle between the light from the light source 210 and the random medium 220. For example, the position driver 230 may be physically coupled to the random medium 220. The position drive device 230 may include fixing means for fixing the random medium 220 with mechanical displacement or percolating means such as a motor or the like. The position driver 230 may move or rotate the random medium 220 in a predetermined manner. For example, the position driver 230 may move the random medium 220 in a predetermined direction and speed over time.

Accordingly, the light emitted from the light source 210 is incident on the random medium 220, and the position of the light incident on the random medium 220 can be changed from the first point to the second point. In addition, the point where the light enters the random medium 220 according to time can be continuously changed.

In another embodiment, the incident position driver 230 may change the position of the light source 210. [ For example, the incident position drive device 230 may change the traveling direction of light emitted from the light source 210 with time. Accordingly, the relative position between the light emitted from the light source 210 and the random medium 220 is changed, and the position of the light incident on the random medium can be changed over time.

In yet another embodiment, the incident position driver 230 may change the angle of the light source 210 to change the angle incident on the light source 210 and the random medium 220. For example, the incident position drive 230 may change the progress angle of light from the light source 210 over time, or the incident drive 230 may rotate the random medium 220 over time The relative angle between the light emitted from the light source 210 and the random medium 220 is changed so that the angle at which the light enters the random medium can be changed with time.

According to the above description, a specific pattern of light having autocorrelation properties can be irradiated onto a fluorescent sample (not shown). In particular, as the relative position or angle between light from the light source 210 and the random medium 220 changes over time, the speckle pattern generated from the random medium 220 also varies over time. The speckle pattern, which changes with time generated from the random medium 220, has auto-correlation characteristics, which will be described in more detail below. Since a speckle pattern that varies with time has an autocorrelation property, even if the fluorescent material of the fluorescent sample (not shown) does not have a blinking phenomenon, an incident speckle pattern is created, and thereby, the effect similar to the flickering of the fluorescent material Lt; / RTI > .

The sensor 240 can sense the light from the random medium 220, the speckle pattern, and the fluorescence signal from the fluorescence sample. Those skilled in the art will readily understand that the sensor 240 is not limited in its type as long as it can sense light such as a photodiode, and the scope of the right is not limited depending on the type of the sensor.

On the other hand, a fluorescent sample (not shown) may be disposed between the random medium 220 and the sensor 240. Light having a speckle pattern from the random medium 220 can be irradiated with a fluorescent sample (not shown), and the sensor 240 can sense a fluorescent signal coming from a fluorescent sample (not shown).

The sensor 240 can convert the sensed light into an electrical signal and output the converted electrical signal to the image processor 250. The sensor 240 may convert the sensed light into an electrical signal at a predetermined time interval. For example, the sensor 240 may sense a speckle pattern generated by scattering light at a first point of the random medium 220 during a first period of time, and may detect a speckle pattern generated during a second period of time, The speckle pattern generated by scattering light at the second point of the random medium 220 changed by the position drive device during the third period without sensing the light while the relative position between the light and the random medium 220 is changed, Lt; / RTI > The sensor 240 can detect the speckle pattern generated by scattering light from the changed position when the position driving device 230 moves the random medium 220 according to time. Or the sensor 240 may sense the speckle pattern generated as the light is scattered from the changed incident point of the random medium as the position drive 230 changes the direction of light from the light source 210 over time. Also, if the random medium 220 or the angle of light is changed with time in the same manner, the speckle pattern generated by scattering light according to the changed incident angle can be detected.

The image processor 250 may generate a plurality of images for the sample over time based on the electrical signals input from the sensor 240. [ For example, the image processor 250 may generate a first image based on an electrical signal input from the sensor 240 during a first period of time. In addition, the image processor 250 may generate a second image based on an electrical signal input from the sensor 240 for a second period of time. The image processor 250 may generate a plurality of images based on an electrical signal input from the sensor 240 for each of a plurality of periods.

The image processor 250 may generate an image, such as a fluorescence image, measured with a general microscope using the mean of a plurality of fluorescence images obtained from the fluorescence sample (not shown). The image processor 250 can generate a high-resolution fluorescence image by applying a predetermined algorithm to a plurality of fluorescence images, which will be described in more detail below.

According to the above, the image generating apparatus 200 can generate a fluorescence image and a high-resolution fluorescence image. In particular, the image generating apparatus 200 according to an exemplary embodiment can generate an effect similar to that using the flickering of a phosphor by irradiating light having a specific pattern without using flicker of the phosphor.

3 is a conceptual diagram for explaining generation of a speckle pattern according to an embodiment.

As shown in FIG. 3, light emitted from a light source (not shown) may be composed of a plurality of rays 301, 302, and 303. Those skilled in the art will readily understand that a plurality of light beams 301, 302, and 303 are only criteria for interpreting light, and are not related to the scope of this right. Hereinafter, it is described that the light is composed of a plurality of light beams.

The plurality of light beams 301, 302, and 303 may propagate through different paths due to scattering within the random medium 310. A plurality of light beams 301, 302 and 303 travel in different paths and a plurality of light beams 301, 302 and 303 traveling in different paths may exit the random medium 310 and generate a speckle pattern 320 according to interference between light rays. The speckle pattern 320 may refer to a random pattern where light is scattered on a rough surface or random medium.

4 is a conceptual diagram for explaining the movement of the random medium and the change of the speckle pattern according to an embodiment. In FIG. 4, since the movement of the random medium changes the relative position between the light incident on the random medium and the random medium, FIG. 4 indicates that the position of the incident light is changed for easy representation in the drawing. In the following, it is described that the position of incident light is changed, but it is emphasized again that the movement is actually a random medium.

As shown in FIG. 4, the position of the light incident on the random medium 220 may move from the first point 211 to the second point 212 by moving the random medium. The first speckle pattern 411 may be generated when the point where the light is incident is the first point 211. [ In addition, if the point at which the light is incident is the second point 212, a second speckle pattern 412 may be generated. On the other hand, it is assumed that the third speckle pattern 413 and the fourth speckle pattern 414 are also generated as the random medium moves with time.

The first speckle pattern 411 to the fourth speckle pattern 414 may be irradiated on the sample 420. [ That is, the fluorescent sample 420 can be irradiated with light having a speckle pattern that varies with time. This is because the fluorescent material flickers due to the speckle pattern in which the fluorescent material existing in the fluorescent sample 420 changes with time, which may be optically similar to the blinking phenomenon of the fluorescent material. For example, when the first speckle pattern 411 is irradiated to the first spot 421 of the fluorescent sample 420 and when the fourth speckle pattern 414 is irradiated to the first spot 421 of the fluorescent sample 420 ), The intensity of light at the first point 421 is changed by the change of the speckle pattern, so that the phosphor flickers with time. That is, when the sample 420 is irradiated with the first speckle pattern 411 to the fourth speckle pattern 414 in this order, the speckle pattern in which the blinking effect is changed in the fluorescent material of the fluorescent sample 420 Lt; / RTI > In particular, the flickering due to the changed speckle pattern at the first point of the fluorescence sample 420 has autocorrelation properties.

That is, the optical characteristics in the fluorescent sample 420 to which the first speckle pattern 411 to the fourth speckle pattern 414 are irradiated may have autocorrelation characteristics. The autocorrelation property can be verified by Equation (1).

Also, although not shown, when the relative angle of the incident light to the random medium 220 is changed, each speckle generated according to the changed angle also has an autocorrelation characteristic. Similar to the above-described relative position change, So that the optical characteristics in the light guide plate 420 can have autocorrelation properties. The autocorrelation property can be verified by Equation (2).

은 광원으로부터 나오는 빛과 랜덤 미디엄의 상대적인 이동량을 의미한다. k는 빛 웨이브 벡터(light wave vector)로써 파장과 관련될 수 있다.

은, 랜덤 미디엄의 평균 자유 행로(mean free path)를 의미한다. 하지만 수학식 1의 경우 상대적인 이동량에 따라 식이 달라지게 된다. 다행히도 이는 빛을 형광샘플에 입사될 때 사용되는 이미징 시스템의 배율(특히, 축소)에 의해 수학식 3과 같이 수정될 수 있다.In Equation (1), r is a coordinate axis indicating the position of the light and the random medium from the light source,

Refers to the amount of relative movement of light and random medium from the light source. k can be related to the wavelength as a light wave vector.

Means the mean free path of the random medium. In Equation (1), however, the equation varies depending on the relative movement amount. Fortunately, this can be modified as shown in equation (3) by the magnification (particularly, reduction) of the imaging system used when light is incident on the fluorescent sample.

는 광원으로부터 나오는 빛과 랜덤 미디엄의 상대적인 각도 변화량을 의미한다. d는 랜덤 미디엄의 두께를 나타낸다. 랜덤 미디엄의 두께와 상대적인 각도를 조절하면 마찬가지로 이미징 시스템의 배율(특히, 축소)에 의해 수학식 3과 같이 수정될 수 있다. In Equation (2), &thetas; denotes the angle between the light emitted from the light source and the random medium,

Refers to the amount of relative angular variation of light and random medium from the light source. d represents the thickness of the random medium. Adjusting the thickness and relative angle of the random medium may likewise be modified as shown in equation (3) by the magnification (especially, reduction) of the imaging system.

은 이미징 시스템의 배율(특히, 축소)에 의해 변형된 좌표축과 상대적인 이동량이며, PSF intensity는 시스템에서 정의된 PSF (point spread function)의 세기이다. 수학식 2의 경우 상대적인 변화량은 각도이나 각도의 변화량이 각도의 변화량이 충분히 작으면 위치의 변화량으로 환산하여 계산할 수 있다. 이 위치의 변화량은 이미징 시스템의 배율(특히, 축소)에 의해 변형되어 그 값이 매우 작아지게 되어 수학식 3으로 변형하여 생각할 수 있다. 이와 같은 이미징 시스템에 의해 수정된 식들은 수학식 3과 같이 하나의 식으로 통일될 수 있으며 증명 과정은 매우 복잡하므로 생략하겠다.In Equation 3, r 'and

PSF intensity is the intensity of the point spread function (PSF) defined in the system. The PSF intensity is the amount of movement relative to the coordinate axis modified by the magnification (particularly, reduction) of the imaging system. In the case of Equation (2), the relative amount of change can be calculated by converting the amount of change in angle or angle into the amount of change in position if the amount of change in angle is sufficiently small. The amount of change of this position is deformed by the magnification (particularly, reduction) of the imaging system, and the value becomes very small. The equations modified by such an imaging system can be unified into one equation as shown in Equation 3, and the verification process is very complicated and will be omitted.

As shown in Equations (1) and (2), flickering occurring in the changed speckle pattern generated by changing the relative position or angle between the light emitted from the light source and the random medium may have an autocorrelation characteristic.

5 is a graph for explaining the resolution enhancement based on the SOFI Fourier reweighting method (FRW).

The method of generating an image according to an exemplary embodiment of the present invention can process a plurality of collected fluorescence images using a statistical method (SOFI) to generate a high-resolution fluorescence image and apply the Fourier aspheric method to the generated high- A fluorescence image with high resolution can be obtained. Road in 5 mean_I the image, such as a fluorescent image measured by the general microscope obtained taking the average of the collected plurality of fluorescence images and the 2 ^nd SOFI is generated by processing images with statistical methods (SOFI) to a plurality of fluorescent images collected The high resolution fluorescence image, 2 ^nd SOFI_FRW, is a fluorescence image with higher resolution produced by applying the Fourier re-weighted method to the resulting high resolution fluorescence image. The graph is a graph showing the intensity of light for each of the displayed distances, and if the resolution prior to the method application is, for example, 533.3 nm, the resolution after the method application is improved to, for example, 346.7 nm.

6 is a conceptual diagram for explaining an image generating apparatus according to an embodiment.

6, the image generating apparatus includes a light source 610, a neutral density filter 611, a random medium 620, a position drive unit 630, a first lens 621, The first mirror 622, the second lens 623, the third lens 624, the first objective lens 625, the object stand 626, the second objective lens 627, the second mirror 628, A fourth lens 629, a fifth lens 631, and a sensor 640. In addition, although not shown, the image generating device may further include an image processor that processes the electrical signals from the sensor 640 to produce an image.

The light source 610 may be, for example, a laser oscillator and may oscillate a laser having a wavelength of 532 nm.

The neutral density filter 611 may be an achromatic filter that does not selectively absorb the output light. The neutral density filter 611 can reduce intensity without changing spectral characteristics with respect to incident light.

The random medium 620 may be a material that can scatter the output light to generate a speckle pattern and may be moved by the position drive device 630. The position driver 630 may move the random medium 620 in a predetermined manner so that the relative position between the light that has exited the light source 610 and passed through the neutral density filter 611 and the random medium 620 Or you can change the angle.

The first lens 621 can be, for example, an FL100 lens and can be used to align light. The first mirror 622 can reflect the incident light and change the traveling direction of the light. The second lens 623 may be an FL150 lens and may be used to align the path of light. The third lens 624 may be an FL200 lens and may be used to align the path of light.

The first objective lens 625 may be a lens used for forming an image of an object. The first objective lens 625 may be, for example, a lens of 50 magnification and 0.5 NA.

The hopper 626 may be a support for placing the sample.

The second objective lens 627 may be a lens used for forming an image of an object. The second objective lens 627 may be, for example, a lens having a magnification of 60 and a refractive index of 0.8 NA.

The second mirror 628 can reflect the light from the second objective lens 627 and change the traveling path.

The fourth lens 629 may be, for example, an FL35 lens and may be used to align the light. The fifth lens 631 may be, for example, an FL500 lens and may be used to align the path of light.

The sensor 640 may sense the incident light and may be, for example, a CCD.

As the light is scattered in the random medium 620, the light irradiated to the fluorescent sample (not shown) on the object table 626 may have a speckle pattern.

In addition, as the position drive unit 630 moves the random medium 620 with time, the speckle pattern irradiated on the fluorescent sample (not shown) may be changed with time. As described above, the speckle pattern irradiating the fluorescent sample (not shown) changes with time, and blinking can be generated in the fluorescent material at plural points in the fluorescent sample (not shown). In addition, flickering at multiple points of the fluorescence sample (not shown) may have autocorrelation properties.

An image processor (not shown) may generate a general fluorescence image and a high-resolution fluorescence image by applying an SOFI image processing method to a plurality of fluorescence images collected by the sensor 640.

7 is an illustration of a plurality of image generation according to an embodiment. Hereinafter, a plurality of speckle patterns are measured and a plurality of speckle patterns are measured to irradiate virtual fluorescence samples to obtain a plurality of fluorescence images.

A plurality of speckle patterns 710 can be generated by moving a random medium over time. For example, although the random medium is shown as moving, it is also possible to change the position where the light emitted from the light source is moved in parallel by the position driving device to enter the random medium. It is also possible to change the relative angle of the random medium or incident light. A plurality of speckle patterns 710 can be irradiated to the fluorescence sample 720. The sensor senses the fluorescence signal from the fluorescence sample 720 and the image processor can generate a plurality of images 730 over time according to the electrical signal from the sensor. The image processor may apply a predetermined algorithm to the plurality of images 730 to generate a normal fluorescence image and a high resolution fluorescence image. The predetermined algorithm here may be an algorithm based on the correlation between adjacent images, and may be, for example, a SOFI method and a fluorescence image generation method by the Fourier aspheric method.

8 is a graph of experimental simulation results according to an embodiment. FIG. 8 shows a fluorescence image using an original pattern, a mean value, and a high-resolution fluorescence image using a second order SOFI FRW method from the left, wherein a fluorescence image using an average value is a fluorescence image obtained using a conventional microscope . It can be seen that the image using the secondary SOFI FRW method has better resolution than the fluorescence image using the average value. In addition, it can be seen that the resolution in the case of applying the secondary SOFI FRW method is also excellent in the position-intensity graph shown below.

9 is a graph for explaining the change of the degree of correlation according to the degree of movement of the random medium.

In FIG. 9, a distance-correlation graph is shown when the degree of movement of the random medium per unit time is 300 nm, 500 nm, 1 mm, and 2 mm. As shown in FIG. 9, by adjusting the degree of movement per unit time of the random medium, the degree of correlation can be controlled.

10 is a block diagram of an apparatus for generating a speckle pattern according to an embodiment.

10, the speckle pattern generating apparatus 1000 may include a random medium 1020 and a position driving apparatus 1030. [

The random medium 1020 may be a disordered matter capable of producing a speckle pattern. The random medium 1020 is a material that can scatter rays of incident light so that rays of incident light travel through various paths so that light passing through the random medium 1020 can have a speckle pattern. There is no.

The position driver 1030 may change the relative position or angle between the incident light and the random medium 1020 by changing the position or angle of the random medium 1020 or directly changing the position or angle of the optical path. For example, the position driver 1030 may be physically connected to the random medium 1020. The position drive 1030 may include mechanical displacement, such as a motor, and fixing means to secure the ramp means and the random medium 1020. The position driver 1030 may move or rotate the random medium 1020 in a predetermined manner. For example, the position driver 1030 may move the position or angle of the random medium 1020 in a predetermined direction and speed. In addition, the position driver 1030 may include a device for changing the path of light incident on the random medium 1020. For example, the position drive device 1030 may change the optical path through a mechanical displacement or a change in angle, such as a Galvano mirror, to set the position or angle at which the light enters the random medium 1020 Direction and speed.

The speckle pattern generating apparatus 1000 can change a relative position with time and generate a speckle pattern having autocorrelation characteristics as described above.

11 is a flow diagram of a method for generating an image for a sample according to an embodiment.

In step 1110, the image generating method may be such that the light output from the light source is directed toward the random medium.

In operation 1120, the incident light is scattered in a random medium to generate light having a speckle pattern, and the generated light can be irradiated to the fluorescent sample.

In step 1130, the image generation method may sense a fluorescent signal emanating from the fluorescent sample.

In step 1140, the image generation method may generate an image based on the sensed fluorescence signal. The image generation method may generate an image for the first period. Here, the first period may mean a period in which the random medium is disposed in the first position.

In step 1150, the image generation method may determine whether image acquisition is terminated. For example, the image generation method may be pre-set to terminate image acquisition once N images have been collected.

If the image acquisition is not terminated, then in step 1160 the image generation method may change the relative position or angle between the incident light and the random medium. For example, the image generation method may change the random medium to the second position. Accordingly, the relative position or angle between the incident light and the random medium can be changed.

The image generation method may return to step 1110 and output light from the light source. The image generation method may repeat steps 1110 to 1140, and may generate an image for the second period.

The image generating method may generate the fluorescence image using the average value of the plurality of images when N images are collected. The image generation method can generate a high-resolution fluorescence image based on the correlation between adjacent images among the N images. For example, the image generation method can generate a high-resolution fluorescence image by applying the algorithm of the SOFI method to the collected N images. The image generation method can apply various order SOFI algorithms, and the order of the SOFI algorithm is not limited.

The apparatus described above may be implemented as a hardware component, a software component, and / or a combination of hardware components and software components. For example, the apparatus and components described in the embodiments may be implemented within a computer system, such as, for example, a processor, a controller, an arithmetic logic unit (ALU), a digital signal processor, a microcomputer, a field programmable array (FPA) A programmable logic unit (PLU), a microprocessor, or any other device capable of executing and responding to instructions. The processing device may execute an operating system (OS) and one or more software applications running on the operating system. The processing device may also access, store, manipulate, process, and generate data in response to execution of the software. For ease of understanding, the processing apparatus may be described as being used singly, but those skilled in the art will recognize that the processing apparatus may have a plurality of processing elements and / As shown in FIG. For example, the processing unit may comprise a plurality of processors or one processor and one controller. Other processing configurations are also possible, such as a parallel processor.

The software may include a computer program, code, instructions, or a combination of one or more of the foregoing, and may be configured to configure the processing device to operate as desired or to process it collectively or collectively Device can be commanded. The software and / or data may be in the form of any type of machine, component, physical device, virtual equipment, computer storage media, or device , Or may be permanently or temporarily embodied in a transmitted signal wave. The software may be distributed over a networked computer system and stored or executed in a distributed manner. The software and data may be stored on one or more computer readable recording media.

The method according to an embodiment may be implemented in the form of a program command that can be executed through various computer means and recorded in a computer-readable medium. The computer-readable medium may include program instructions, data files, data structures, and the like, alone or in combination. The program instructions to be recorded on the medium may be those specially designed and configured for the embodiments or may be available to those skilled in the art of computer software. Examples of computer-readable media include magnetic media such as hard disks, floppy disks and magnetic tape; optical media such as CD-ROMs and DVDs; magnetic media such as floppy disks; Magneto-optical media, and hardware devices specifically configured to store and execute program instructions such as ROM, RAM, flash memory, and the like. Examples of program instructions include machine language code such as those produced by a compiler, as well as high-level language code that can be executed by a computer using an interpreter or the like. The hardware devices described above may be configured to operate as one or more software modules to perform the operations of the embodiments, and vice versa.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments. For example, it is to be understood that the techniques described may be performed in a different order than the described methods, and / or that components of the described systems, structures, devices, circuits, Lt; / RTI > or equivalents, even if it is replaced or replaced.

Therefore, other implementations, other embodiments, and equivalents to the claims are also within the scope of the following claims.

Claims (8)

A light source for outputting light;
A random medium for scattering the output light to generate light having a speckle pattern;
A position drive device for generating a speckle pattern varying in time with an auto correlation characteristic by changing a relative position or an angle between light incident on the random medium and the random medium outputted from the light source;
A sensor for sensing a fluorescence signal from the fluorescent sample and converting the fluorescence signal into an electrical signal when light having the speckle pattern is irradiated to the fluorescent sample; And
An image processor for obtaining a plurality of images according to relative positions changed with time of the random medium by using the converted electrical signals and processing the plurality of images to generate a fluorescent image,
And an image generating device. The method according to claim 1,
Wherein the image processor is configured to apply a super-resolution optical fluctuation imaging (SOFI) method to the plurality of images based on a degree of correlation between adjacent images of the plurality of images, and to generate a high- . 3. The method of claim 2,
The image processor comprising:
A plurality of images are processed by the SOFI method to generate a fluorescence image having a first resolution, and a Fourier reweighting method (FRW) is applied to the fluorescence image having the first resolution, Image,
Wherein the second resolution is higher than the first resolution,
Image generating device. The method according to claim 1,
The position drive apparatus moves at least one of a position where the light output from the light source is incident on the random medium and the random medium to change a relative position between the incident light and the random medium, Changing,
Image generating device. The method of claim 1, wherein
Wherein the position drive device changes at least one of an angle of a light incident on the random medium and a random medium angle of the light output from the light source to change a relative angle between the incident light and the random medium, Changing the pattern,
Image generating device. The method according to claim 1,
Wherein the position drive device adjusts the degree of change in relative position or angle between the incident light and the random medium per unit time. A random medium for generating a speckle pattern while transmitting incident light; And
A driving device for generating a speckle pattern having an auto-correlation characteristic and changing with time by changing a relative position between the incident light and the random medium,
The spark pattern generating apparatus comprising: Outputting light such that light having a speckled pattern scattered in a random medium is irradiated on the sample;
Sensing a fluorescence signal from the fluorescent sample to produce an image;
Changing a relative position or angle between light incident on the random medium and the random medium according to time to generate a speckle pattern having an auto correlation characteristic and changing with time;
Generating and collecting images using light having a speckle pattern varying with time; And
Processing the plurality of images to produce a fluorescence image
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