KR20140075450A - 3d imaging modeling method using fluorescent intensity profile analysis - Google Patents

3d imaging modeling method using fluorescent intensity profile analysis Download PDF

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Publication number
KR20140075450A
KR20140075450A KR1020120143799A KR20120143799A KR20140075450A KR 20140075450 A KR20140075450 A KR 20140075450A KR 1020120143799 A KR1020120143799 A KR 1020120143799A KR 20120143799 A KR20120143799 A KR 20120143799A KR 20140075450 A KR20140075450 A KR 20140075450A
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South Korea
Prior art keywords
phosphor
fluorescence signal
intensity profile
information
fluorescent
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KR1020120143799A
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Korean (ko)
Inventor
정은주
김봉규
장원익
송현우
안창근
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한국전자통신연구원
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Priority to KR1020120143799A priority Critical patent/KR20140075450A/en
Publication of KR20140075450A publication Critical patent/KR20140075450A/en

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    • G—PHYSICS
    • G06—COMPUTING; CALCULATING OR COUNTING
    • G06T—IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
    • G06T15/00—3D [Three Dimensional] image rendering
    • G06T15/06—Ray-tracing
    • G—PHYSICS
    • G06—COMPUTING; CALCULATING OR COUNTING
    • G06T—IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
    • G06T2207/00—Indexing scheme for image analysis or image enhancement
    • G06T2207/10—Image acquisition modality
    • G06T2207/10064—Fluorescence image

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  • Engineering & Computer Science (AREA)
  • Computer Graphics (AREA)
  • Physics & Mathematics (AREA)
  • General Physics & Mathematics (AREA)
  • Theoretical Computer Science (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)

Abstract

A three-dimensional image modeling method that can be implemented as a three-dimensional image through fluorescence signal intensity profile analysis is disclosed. A three-dimensional image modeling method for this purpose includes a fluorescent signal acquisition step of acquiring a fluorescence signal from a fluorescent substance by irradiating light emitted from a light source to a sample having at least one fluorescent substance, a fluorescent light converting the obtained fluorescent signal into a fluorescent signal intensity profile A step of converting the signal intensity profile, a step of calculating the phosphor information to calculate the depth information, the position information and the size information of the phosphor incorporated in the sample by analyzing the shape change of the fluorescence signal intensity profile, and the step of calculating the depth information, Dimensional image by using a three-dimensional image.

Description

FIELD OF THE INVENTION [0001] The present invention relates to a 3D image modeling method using fluorescence signal intensity analysis,

A three-dimensional image modeling method, and more particularly, a three-dimensional image modeling method using fluorescence image are disclosed.

The bio-optical imaging device is intended to analyze biological processes at the cell or molecular level in a live state and to analyze the biological processes in the living body. The object to be imaged is a biological change at a molecular level. Fluorescence imaging acquires fluorescence in the range of visible light and imaged. Non-invasive methods can be used to observe changes in cells inside and outside the body in real time. Recently, research has been conducted to apply fluorescence technology to the human body to develop medical devices. .

Confocal Laser Scanning Microscope can be used for non-destructive observation of the optical and monolayer structures of cells and tissues. It is possible to observe the position and morphology of the material. Can be created. By scanning the sample surface with a laser beam, only the light of the focal plane is detected, and only the focused image is obtained. At the same time, the space information is recorded, and the slice image is reproduced through the computer. As a result, the sample is optically sectioned on the Z-axis (depth direction) to obtain a clear optical slice.

In this case, it is advantageous to obtain a high-quality image. However, since only one spot is focused on, a large amount of image information about the segment at each focus is required in order to implement the entire three-dimensional image.

US registration number 8110405 (2012.02.07)

Dimensional image modeling method that can be implemented as a three-dimensional image through analysis of a fluorescence signal intensity profile.

According to one aspect, a three-dimensional image modeling method includes a fluorescent signal obtaining step of obtaining a fluorescent signal from a fluorescent substance by irradiating light emitted from a light source onto a sample having at least one fluorescent substance, A fluorescence signal intensity profile conversion step of converting the fluorescence intensity profile of the fluorescent material, a fluorescent signal intensity profile conversion step of converting the fluorescence intensity profile of the fluorescence signal, a depth information of the fluorescent material embedded in the sample, And a 3D image implementation step of implementing the 3D image using the size information.

The disclosed three-dimensional image modeling method can realize a three-dimensional image at a living tissue level using a less amount of image information through analysis of a fluorescence signal intensity profile.

The disclosed 3D image modeling method can visualize the shape and internal structure of the living body without any damage to the sample in a noncontact, real-time, non-incisional manner, and can analyze the three-dimensional structure and microstructure.

1 is a flowchart of a 3D image modeling method according to an exemplary embodiment of the present invention.
FIGS. 2A and 2B are views for explaining the acquisition of the fluorescence signal and the conversion of the fluorescence signal intensity profile when the phosphor according to one embodiment is located at a shallow position in the sample.
FIGS. 3A and 3B are diagrams for explaining the acquisition of the fluorescence signal and the switching of the fluorescence signal intensity profile when the phosphor according to one embodiment is at a deep position in the sample.
4A and 4B are views for explaining calculation of phosphor information when two phosphors according to one embodiment are in a sample.
5 is a block diagram of a 3D image modeling system according to an embodiment.

The foregoing and further aspects of the invention will become apparent through the following examples. The configurations of the selectively described embodiments or selectively described embodiments of the present invention may be freely combined with each other if they are not explicitly contradictory to those of ordinary skill in the art, I understand.

1 is a flowchart of a 3D image modeling method according to an exemplary embodiment of the present invention.

A three-dimensional image modeling method includes a fluorescence signal acquisition step (S110) of acquiring a fluorescence signal from a fluorescent material by irradiating light emitted from a light source onto a sample having at least one fluorescent material, a fluorescent light intensity profile obtaining step A signal intensity profile switching step (S130), a phosphor information calculation step (S150) of calculating depth information, position information and size information of the phosphor embedded in the sample by analyzing the change in the shape of the fluorescence signal intensity profile, Dimensional image using the position information and the size information (S170).

In the fluorescence signal acquisition step (S110), the fluorescence signal can be acquired from the fluorescent material by irradiating the sample containing at least one fluorescent material with light output from the light source. When the light emitted from the light source is irradiated on the sample, the fluorescent substance in the sample absorbs light of a specific wavelength, and then can emit light having a wavelength inherent to the fluorescent substance. The shape or distribution of the phosphor according to the light emitted from the phosphor can be measured using a CCD camera. Details of the acquisition of the fluorescence signal will be described later with reference to Figs. 2A and 3A.

In the fluorescence signal intensity profile switching step S130, a fluorescence signal intensity profile can be obtained from the acquired fluorescence signal image. The image of the obtained fluorescence signal can be read and a value proportional to the intensity of the fluorescence signal can be calculated and converted into a fluorescence signal intensity profile. Concrete contents of the switching of the fluorescence signal intensity profile will be described later with reference to Figs. 2B and 3B.

In the phosphor information calculation step (S150), depth information, position information, and size information of the phosphor embedded in the sample can be calculated by analyzing the shape change of the fluorescence signal intensity profile. Using the fact that the shape of the fluorescence signal intensity profile changes according to the depth of the fluorescent substance in the sample, the depth information of the fluorescent substance can be obtained by analyzing the shape change of the fluorescent signal intensity profile. Details of the phosphor information calculation will be described later with reference to Figs. 4A and 4B.

In the 3D image implementation step (S170), a three-dimensional image can be realized by using the depth information, position information, and size information of the calculated phosphor. The shape of the phosphor in the sample can be three-dimensionally represented using depth information, position information, and size information of the phosphor. Thus, by analyzing the change in the shape of the profile relative to the fluorescence signal intensity of the phosphor, information on depth, position, and size can be obtained, and a three-dimensional stereoscopic fluorescence image can be obtained with a surface area of several mm.

FIGS. 2A and 2B are views for explaining the acquisition of the fluorescence signal and the conversion of the fluorescence signal intensity profile when the phosphor according to one embodiment is located at a shallow position in the sample.

Phosphors are substances that absorb energy and emit fluorescence. Phosphors include intrinsic fluorescent materials, which have their own fluorescent properties, and extrinsic fluorescent materials, which combine with the object to be analyzed to make the object to be analyzed fluoresce.

According to one embodiment, the fluorescent substance 251 in the sample 240 may be a self fluorescent substance having a fluorescent property as an object to be analyzed. According to one embodiment, when an object to be analyzed does not have a specific fluorescent property or has a fluorescent property that is not suitable for the experiment, fluorescence can be measured by binding an external fluorescent substance useful for analysis to an analyte.

According to one embodiment, a uniform phosphor can be used to more accurately obtain the information of the phosphor. In this case, when the absorbance, the size, the phosphor concentration, and the like, which are information related to the luminous efficiency of the phosphor, are used, the amount of light emitted from the phosphor can be accurately known. In the case where a large amount of the phosphor is distributed, .

A known laser scanning microscope can be used for observation of the sample. Since the structure of the laser scanning microscope is well known, the remaining configuration of the laser scanning microscope except for the CCD (Charged Couple Device) camera 210, the color filter 220 and the light source 230 is omitted in FIG. .

When the light output from the light source 230 is irradiated to the sample 240, the sample fluorescent material 251 absorbs light of a specific wavelength and then emits light having a wavelength specific to the fluorescent material. The shape or the distribution of the phosphor according to the light emitted from the phosphor can be measured using the CCD camera 210. The color filter 220 minimizes the incidence of the light emitted from the light source 230 to the sample 240 into the CCD camera 210, thereby enhancing the fluorescence image characteristic measured.

The light emitted from the fluorescent material 251 embedded in the sample 240 may pass through the sample 240, and the light may gradually spread due to the scattering phenomenon. When the fluorescent substance 251 is present near the surface of the sample 240, the fluorescence emitted from the fluorescent substance 251 may have a small scattering effect and a small spread of light. Therefore, as shown in FIG. 2B, a narrow range of the strong fluorescence signal image 311 can be obtained in the CCD camera 210. FIG.

The conversion from the fluorescence signal image to the fluorescence signal intensity profile can be performed by reading the fluorescence intensity value in the fluorescence image as shown in FIG. 2B and indicating the magnitude of the fluorescence signal intensity according to the fluorescence intensity value. Fluorescent images at the acquired focus can be read sequentially from the top to switch to the fluorescence signal intensity profile.

When the fluorescent substance 251 is present near the surface of the sample 240, the fluorescence signal intensity profile 321, which is converted from the fluorescent signal image obtained at the illustrated arrow position, may appear close to a square shape. The fluorescence signal intensity profile can be obtained from the top in the direction of the arrow sequentially from the top of the fluorescence image. When the depth information of the phosphor is calculated using the fluorescence signal intensity profile in order to realize a three-dimensional image, the depth of the phosphor in the sample may be calculated to be shallow as it is closer to the square shape.

FIGS. 3A and 3B are diagrams for explaining the acquisition of the fluorescence signal and the switching of the fluorescence signal intensity profile when the phosphor according to one embodiment is at a deep position in the sample.

 3A, when the phosphor 252 is located at a deep position in the sample 240 according to an embodiment, the fluorescence emitted from the phosphor 252 generates a large scattering effect, . Therefore, as shown in FIG. 3B, a wide range of weak fluorescence signal images 312 can be obtained in the CCD camera 210. [

When the phosphor 252 is in a deep position in the sample 240, the fluorescence signal intensity profile 322, which is switched from the fluorescence signal image obtained at the illustrated arrow position, may appear in a gentle curve. The fluorescence signal intensity profile can be obtained from the top in the direction of the arrow sequentially from the top of the fluorescence image. When the depth information of the phosphor is calculated using the fluorescence signal intensity profile in order to implement a three-dimensional image, the depth of the phosphor in the sample can be calculated as being closer to a gentle curve shape.

4A and 4B are views for explaining calculation of phosphor information when two phosphors according to one embodiment are in a sample.

4A, when two phosphors 253 and 254 having different depths exist in the sample 240 according to one embodiment, the fluorescence signal image 313 obtained by the CCD camera 210 is May be the same as shown in FIG. 4B. A strong fluorescence signal image in a narrow range by the fluorescent substance 253 at the shallow position and a weak fluorescence signal image in a wide range by the fluorescent substance 254 at the deep position may overlap.

The overall fluorescence signal intensity profile 325 is switched to a form in which the fluorescent signal intensity profile 323 by the fluorescent substance 253 in the shallow position and the fluorescence signal intensity profile 324 by the fluorescent substance 254 in the deep position are overlapped . The depth information of the phosphor can be obtained by using the property that the shape of the fluorescence signal intensity profile is different depending on the position of the phosphor. When the shape change of the fluorescence signal intensity profile 325 is analyzed, the depth information of the phosphor at the deep position on the left side in the gentle curve form on the left side and the depth information of the phosphor in the shallow position on the right side in the high curve shape on the right side are respectively calculated .

The positional information of the phosphor may indicate the position of the phosphor along the x and y axes in a plane perpendicular to the z axis, where depth is the z axis. It is possible to calculate the positional information of the phosphor with respect to an arbitrary reference point on the sample by analyzing the fluorescence signal image. The size information of the phosphor can be obtained by using the depth information and the position information of the calculated phosphor.

The shape of the phosphor in the sample can be three-dimensionally represented using depth information, position information, and size information of the phosphor. Thus, by analyzing the change in the shape of the profile relative to the fluorescence signal intensity of the phosphor, information on depth, position, and size can be obtained, and a three-dimensional stereoscopic fluorescence image can be obtained with a surface area of several mm.

In case of FIG. 4A, it is assumed that there are two phosphors in the sample. As described above, analysis of the morphological change of the fluorescence signal intensity profile and implementation of the three-dimensional image can be applied as it is even when there are three or more phosphors in the sample. The arrows shown in FIGS. 2B, 3B, and 4B indicate the direction of reading the fluorescent image according to one embodiment, and the present invention is not limited thereto.

5 is a block diagram of a 3D image modeling system according to an embodiment.

The three-dimensional image modeling system 500 includes a fluorescence signal acquisition unit 510 for acquiring a fluorescence signal from a fluorescent material by irradiating light from a light source to a sample having at least one fluorescent material therein, And a 3-dimensional image implementing unit 520 for calculating information of the phosphor by analyzing the change in shape of the fluorescence signal intensity profile and implementing the 3-dimensional image using the information of the calculated phosphor.

In the case of the fluorescence signal acquisition unit 510, the fluorescence signal can be obtained from the fluorescent material by irradiating the sample containing at least one fluorescent material with light output from the light source. According to one embodiment, the fluorescence signal acquisition unit 510 may be a known laser scanning microscope.

The three-dimensional image realization unit 520 can send a signal to the fluorescence signal acquisition unit 510 regarding the irradiation position of the light source, the control of the CCD camera, and the like. The fluorescence signal image obtained by the fluorescence signal acquisition unit 510 may be transmitted to the three-dimensional image implementation unit 520. In the three-dimensional image realization unit 520, the transmitted fluorescence signal image can be read and converted into a fluorescence signal intensity profile according to a previously stored program or the like.

Using the fact that the shape of the fluorescence signal intensity profile changes according to the depth of the fluorescent substance in the sample, the three-dimensional image implementing unit 520 analyzes the shape change of the converted fluorescence signal intensity profile through a pre- Information can be obtained. According to an exemplary embodiment, when a plurality of phosphors are present in the sample, depth information of each phosphor can be calculated by analyzing the overlapping characteristics of the plurality of fluorescent signal intensity profiles corresponding to each of the plurality of phosphors.

The positional information of the phosphor may indicate the position of the phosphor along the x and y axes in a plane perpendicular to the z axis, where depth is the z axis. It is possible to calculate the positional information of the phosphor with respect to an arbitrary reference point on the sample by analyzing the fluorescence signal image. The size information of the phosphor can be obtained by using the depth information and the position information of the calculated phosphor.

In the 3D image implementation unit 520, at least one phosphor may be modeled as a three-dimensional image according to depth information, position information, and size information of the phosphor calculated using the pre-stored program. The shape of the phosphor in the sample can be three-dimensionally represented using depth information, position information, and size information of the phosphor.

By analyzing the profile shape change of the fluorescent signal intensity of the phosphor, information on the depth, position and size can be obtained, and it can be realized as a three-dimensional stereoscopic fluorescence image with a surface area of several mm. According to one embodiment, the three-dimensional image implementation unit 520 may be implemented by a computing device capable of embedding a computer or other program.

As described above, in the 3D image modeling system 500, the distribution of the fluorescence signal intensity profile is analyzed by software and the depth information is measured, and the observation object is three-dimensionally stereoscopically imaged using the position information and the size information, Can be analyzed.

It will be apparent to those skilled in the art that various modifications, substitutions and substitutions are possible, without departing from the scope and spirit of the invention as disclosed in the accompanying claims. will be. Therefore, the embodiments disclosed in the present invention and the accompanying drawings are intended to illustrate and not to limit the technical spirit of the present invention, and the scope of the technical idea of the present invention is not limited by these embodiments and the accompanying drawings . The scope of protection of the present invention should be construed according to the following claims, and all technical ideas falling within the scope of the same shall be construed as falling within the scope of the present invention.

210: CCD camera 220: Color filter
230: light source 240: sample
251, 252, 253, and 254:
311, 312, 313: Fluorescence signal image
321, 322, 323, 324, 325: Fluorescence signal intensity profile
500: 3D image modeling system
510: Fluorescence signal acquisition unit 520: 3D image implementation unit

Claims (1)

A fluorescent signal obtaining step of obtaining a fluorescence signal from a fluorescent material by irradiating light emitted from a light source to a sample having at least one fluorescent substance;
A fluorescent signal intensity profile switching step of converting the obtained fluorescence signal into a fluorescence signal intensity profile;
A phosphor information calculation step of calculating depth information, position information, and size information of the fluorescent material contained in the sample by analyzing the shape change of the fluorescent signal intensity profile; And
A three-dimensional image realizing step of realizing a phosphor as a three-dimensional image using depth information, position information, and size information of the calculated phosphor;
Dimensional image modeling method.
KR1020120143799A 2012-12-11 2012-12-11 3d imaging modeling method using fluorescent intensity profile analysis KR20140075450A (en)

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20180062411A (en) * 2016-11-30 2018-06-08 단국대학교 산학협력단 Method of acquiring an image of a biological sample
KR20190051506A (en) 2017-11-07 2019-05-15 주식회사 에프아이시스 Multi-angle Image Acquisition System
CN117379007A (en) * 2023-12-07 2024-01-12 合肥锐视医疗科技有限公司 3D optical imaging system and method

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20180062411A (en) * 2016-11-30 2018-06-08 단국대학교 산학협력단 Method of acquiring an image of a biological sample
KR20190051506A (en) 2017-11-07 2019-05-15 주식회사 에프아이시스 Multi-angle Image Acquisition System
CN117379007A (en) * 2023-12-07 2024-01-12 合肥锐视医疗科技有限公司 3D optical imaging system and method
CN117379007B (en) * 2023-12-07 2024-03-15 合肥锐视医疗科技有限公司 3D optical imaging system and method

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