KR20170038613A - Apparatus and method for image processing - Google Patents

Apparatus and method for image processing Download PDF

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KR20170038613A
KR20170038613A KR1020150160492A KR20150160492A KR20170038613A KR 20170038613 A KR20170038613 A KR 20170038613A KR 1020150160492 A KR1020150160492 A KR 1020150160492A KR 20150160492 A KR20150160492 A KR 20150160492A KR 20170038613 A KR20170038613 A KR 20170038613A
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cells
analyzed
blood vessel
living body
cell
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KR101831820B1 (en
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김필한
서호원
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한국과학기술원
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00—Measuring for diagnostic purposes; Identification of persons
    • A61B5/0059—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence
    • A61B5/0062—Arrangements for scanning
    • A61B5/0068—Confocal scanning
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00—Measuring for diagnostic purposes; Identification of persons
    • A61B5/145—Measuring characteristics of blood in vivo, e.g. gas concentration, pH value; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid, cerebral tissue
    • A61B5/1455—Measuring characteristics of blood in vivo, e.g. gas concentration, pH value; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid, cerebral tissue using optical sensors, e.g. spectral photometrical oximeters
    • A61B5/14551—Measuring characteristics of blood in vivo, e.g. gas concentration, pH value; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid, cerebral tissue using optical sensors, e.g. spectral photometrical oximeters for measuring blood gases
    • A61B5/14556—Measuring characteristics of blood in vivo, e.g. gas concentration, pH value; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid, cerebral tissue using optical sensors, e.g. spectral photometrical oximeters for measuring blood gases by fluorescence
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5094—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for blood cell populations
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
    • G01N33/582—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with fluorescent label
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/36—Microscopes arranged for photographic purposes or projection purposes or digital imaging or video purposes including associated control and data processing arrangements
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B2207/00—Coding scheme for general features or characteristics of optical elements and systems of subclass G02B, but not including elements and systems which would be classified in G02B6/00 and subgroups
    • G02B2207/113—Fluorescence

Abstract

An image processing apparatus according to an embodiment of the present invention includes an image capturing unit for capturing a blood vessel at a specific position and outputting an image signal, And calculating a number of cells to be analyzed circulated in vivo using the number of reference cells and the number of cells to be analyzed.

Description

[0001] APPARATUS AND METHOD FOR IMAGE PROCESSING [0002]

The present invention relates to an apparatus and method for image processing of living circulatory cells.

The method of extracting blood from the blood and analyzing the blood cancer cells present in the bloodstream and the cells expressing the specific protein is widely used for early diagnosis of cancer metastasis and early diagnosis of specific diseases.

However, it has disadvantages in that it is limited in terms of sensitivity and accuracy in detecting a very small amount of cells in the bloodstream. Therefore, studies are underway to analyze the blood cancer cells and the cells expressing the specific protein present in the blood by directly sensing the circulating cells in the body without extracting the blood outside the body.

There is a need for a method and apparatus for imaging circulating cells in the blood to accurately determine the presence or absence of diseases such as progression or recurrence of cancer metastasis and quantifying circulating cells present in the blood based on the images.

In this case, since a part of blood vessels is photographed for a predetermined time to detect cells circulating in the blood, there is a limit in that it is difficult to quantify circulating cells in the entire blood inside the living body.

An object of the present invention is to provide an image processing apparatus and a method for image processing of a living circulatory cell capable of capturing a part of a blood vessel to detect circulating cells and quantifying circulating cells in the whole blood.

An image processing apparatus according to an embodiment of the present invention includes a photographing unit for photographing a blood vessel at a specific position and outputting an image signal, a control unit for acquiring the number of reference cells and analysis target cells sensed from the image signal, And an operation unit for converting the number of cells to be analyzed circulated in the living body by using the number of cells to be analyzed.

Wherein the calculation unit calculates a calibration coefficient using the number of reference cells injected into the living body and the number of reference cells sensed in the blood vessel and calculates a calibration coefficient by using the calibration coefficient and the number of cells to be analyzed, It is possible to estimate the number of cells to be analyzed circulating in the cell.

The calibration coefficient may be a ratio of reference cells passing through the blood vessel among the reference cells injected into the living body.

The number of reference cells detected in the blood vessel may be an average value of the number of reference cells calculated in each of a plurality of image signals of the blood vessels.

The calculation unit may convert the number of cells to be analyzed circulated in the living body from the number of cells to be analyzed detected in the blood vessel using the calibration coefficient.

The reference cell may be a cell that continuously circulates in vivo.

The analyte cell may be a cell labeled with a first fluorescent sample, and the reference cell may be a cell labeled with a second fluorescent sample.

The imaging unit includes a light irradiation unit for outputting at least one laser light source, a lens unit for adjusting the focus of the laser light source in the blood vessel, a beam scanning unit for reading the image observed from the blood vessel and generating a two- And a video signal sensing unit for generating an image including a fluorescent material excited by the laser light source by passing the scanning image through a pinhole provided on a confocal plane surface.

The image processing method according to an embodiment of the present invention includes the steps of storing the number of cells to be analyzed and reference cells injected into a living body, photographing a specific blood vessel, analyzing the captured image, And calculating the number of cells to be analyzed in the living body by using the number of reference cells and the number of cells to be analyzed.

The step of converting the number of cells to be analyzed circulating in the living body may include calculating a calibration coefficient using the number of reference cells injected into the living body and the number of reference cells sensed by the blood vessel, And estimating the number of cells to be analyzed circulated in the living body using the number of cells to be analyzed detected in the blood vessel.

The step of calculating the calibration coefficient may be determined as a ratio of the reference cells detected in the blood vessel among the reference cells injected into the living body.

The number of reference cells detected in the blood vessel may be an average value of the number of reference cells calculated in each of a plurality of image signals of the blood vessel.

The step of estimating the number of cells to be analyzed circulated in the living body may be estimated by multiplying the calculated calibration coefficient by the number of cells to be analyzed detected in the blood vessel.

The reference cell may be a cell that continuously circulates in vivo.

The analyte cell may be a cell labeled with a first fluorescent sample, and the reference cell may be a cell labeled with a second fluorescent sample.

According to an embodiment of the present invention, cells in organs circulating in the organism can be detected and quantified.

1 is a block diagram of an image processing apparatus according to an embodiment of the present invention.
FIG. 2 is a view showing analyzed cells detected from an image obtained according to an embodiment of the present invention. FIG.
FIG. 3 is a graph illustrating the number of cells to be analyzed according to an embodiment of the present invention.
FIG. 4 is a graph illustrating the number of cells to be analyzed according to an embodiment of the present invention.
FIG. 5 is a photograph showing major organs after the cells to be analyzed are injected.
6 is a view for explaining a photographing unit according to an embodiment of the present invention.
7 is a view showing reference cells and analyzed cells detected from images obtained according to an embodiment of the present invention.
8 is a graph showing the number of reference cells and analyzed cells detected from an image obtained according to an embodiment of the present invention according to the flow of time and the number of detected reference cells and the number of cells to be analyzed FIG. 2 is a graph showing an average value obtained by the following equation.
9 is a flowchart of an image processing method according to an embodiment of the present invention.

Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings so that those skilled in the art can easily carry out the present invention. The present invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. In order to clearly illustrate the present invention, parts not related to the description are omitted, and similar parts are denoted by like reference characters throughout the specification.

Throughout the specification, when an element is referred to as "comprising ", it means that it can include other elements as well, without excluding other elements unless specifically stated otherwise.

FIG. 1 is a block diagram of an image processing apparatus according to an embodiment of the present invention. FIG. 2 is a diagram showing cells to be analyzed detected from an image obtained according to an embodiment of the present invention. FIG. 4 is a graph illustrating the number of cells to be analyzed according to an embodiment of the present invention, and FIG. 4 is a graph illustrating the number of cells to be analyzed according to an embodiment of the present invention. 5 is a photograph of major organs after injecting the cells to be analyzed.

The image processing apparatus 100 according to an embodiment of the present invention photographs an image of a circulatory cell in a living body using a confocal microscope and processes the photographed image. Based on the rotation angle of 36 or 72 angles, Designed to quickly obtain high-resolution images.

Referring to FIG. 1, an image processing apparatus 100 according to an embodiment of the present invention includes a photographing unit 200 and an operation unit 300.

The photographing unit 200 photographs a specific blood vessel and acquires an image of the analysis target cell labeled with a fluorescent sample circulating in vivo as shown in FIG.

A cell to be analyzed is a blood cancer cell or a cell in which a specific protein is expressed in the bloodstream, and means a cell to be detected. In the present embodiment, the cells to be analyzed may be colon cancer cells, but the present invention is not limited thereto. Cells to be analyzed may be labeled with a first fluorescent sample.

In this embodiment, the photographing unit 200 can photograph the Great Saphenous Vein (GSV) to fix the blood vessels to minimize dynamic noise and to analyze a large amount of blood.

The arithmetic unit 300 may sort the image of the blood vessel acquired by the photographing unit 200, and may detect the fluorescence-labeled cells to be analyzed and count the number of the cells.

However, referring to FIG. 3, it can be seen that the number of cells to be analyzed reaches a peak within 2 minutes after the cells to be analyzed are injected, and thereafter, the number of cells to be analyzed is remarkably reduced. In particular, referring to FIG. 4, it can be seen that the number of cells to be analyzed having a size of less than 7.5 .mu.m among the cells to be analyzed is significantly decreased with time. This is because the cells to be analyzed having a size of less than 7.5 .mu.m remain in a specific organ as shown in FIG. 5 while circulating the blood vessels, thereby reducing the number of cells to be analyzed for long-term circulation of blood vessels.

As described above, since a part of the analyte cells having a size smaller than a certain size remains in organs and does not circulate in the blood vessels for a long time, a part of the blood vessels is photographed for a predetermined time to detect cells circulating in the blood. It is very difficult to quantify circulating cells in the whole blood inside.

The image processing apparatus 100 of the present invention injects the reference cells and the cells to be analyzed, which will be described below, into the same number, and quantifies the cells to be analyzed circulating in the whole blood using the number of reference cells detected in the blood vessels .

Reference cells are cells that circulate continuously in the blood vessels, and they use circulating cells that are generally not fragmented even though time passes. In the present embodiment, red blood cells are used as the reference cells, but the present invention is not limited thereto. Any cell (for example, nanoparticles or the like) that circulates intravascularly without remaining in the body can be used. The reference cell is labeled with a second fluorescent sample and is distinguished from the analyte cell.

Referring again to FIG. 1, the photographing unit 200 acquires an image of a reference cell labeled with the first fluorescent sample and a reference cell labeled with the second fluorescent sample.

FIG. 6 is a view for explaining a photographing unit according to an embodiment of the present invention, and FIG. 7 is a diagram showing analyzed cells and reference cells sensed from an image obtained according to an embodiment of the present invention.

The photographing unit 200 includes a light irradiating unit 210, a lens unit 220, a beam scanning unit 230, and an image signal sensing unit 240.

The light irradiation unit 210 irradiates light to excite a fluorescent substance labeled with cells in a blood vessel as a light source of the photographing unit 200. In this embodiment, the light irradiating unit 210 is composed of three laser light sources having a visible light band, and the laser light sources may have wavelength bands of 488 nm, 561 nm, and 640 nm, respectively. However, the number and the wavelength of the light source may be variously changed.

The lens unit 220 allows the light irradiated by the light irradiation unit 210 to focus on the blood vessel. Also, as a lens through which the light of the fluorescent material excited by the light irradiation unit 210 enters, a video signal having an image of a cell labeled with a fluorescent substance is output to the beam scanning unit 230.

At this time, in this embodiment, the lens unit 220 is rotated at 250 x 250

Figure pat00001
, Or using a 20x objective lens to have a field of view of 500x500
Figure pat00002
However, the present invention is not limited to this, and it may be set to have various fields of view by setting a lens having an appropriate magnification.

The beam scanning unit 230 reads a video signal incident through the lens unit 220 and constructs a pixel in a two-dimensional array form. At this time, the beam scanning unit 230 may include a polygonal rotation mirror, and a galvanometer mirror. The rotating polygon mirror scans the X-line, and the Galvano mirror scans the Y-line.

The image signal sensing unit 240 generates an image including a fluorescent material by passing a scanning image generated from the beam scanning unit 230 through a pinhole provided on a confocal plane. The image signal sensing unit 240 includes a beam splitter, a band pass filter (BPF), a photomultiplier tube (PMT), and a frame grabber.

The beam splitter separates and arranges the scanning image generated from the beam scanning unit 230, and may be a Dichroic Beam Split (DBS). In this embodiment, the beam splitter can separate and align the excited video signal into three primary colors.

A band pass filter (BPF) is located in a path of light divided from a beam splitter to obtain separated light and pass light in a spectral range of a designated visible light region.

The photomultiplier tube is located in the path of the light passed from the band-pass filter section, detects the fluorescent signal passing through the band-pass filter section, and generates an electric signal.

The frame grabber digitizes an electric signal formed from the photoelectron multiplier and outputs the digitized signal to the arithmetic unit 300.

As shown in FIG. 7, the operation unit 300 can detect the first cell and the second cell by sensing the first fluorescent material and the second fluorescent material from the image signal obtained from the photographing unit 200.

At this time, the operation unit 300 converts the number of cells to be analyzed in the living body by using the analyzed cells and the reference cells detected in the blood vessel after injecting the analyzed cells and the reference cells into the body.

The calculation unit 300 obtains a calibration coefficient C that can be converted into the number of cells circulating in the whole blood using the number of reference cells injected in vivo and the number of reference cells sensed in the blood vessel.

The calibration coefficient (C) can be defined as shown in Equation (1).

Figure pat00003

Figure pat00004
Is the number of reference cells injected into the living body,
Figure pat00005
Is the number of reference cells detected in the blood vessel.

At this time, the number of reference cells detected in blood vessels

Figure pat00006
Can count the number of reference cells calculated in each of a plurality of image signals of a blood vessel, and can further improve the reliability by using a value obtained by calculating the average of these reference cells.

Next, the calculation unit 300 calculates the number of cells to be analyzed circulating in the whole blood using the calculated correction coefficient C and the number of cells to be analyzed detected in the blood vessel.

The number of cells to be analyzed circulating in the whole blood can be calculated as shown in Equation (2).

Figure pat00007

Figure pat00008
Is the number of cells to be analyzed circulating in the whole blood,
Figure pat00009
Is the number of cells analyzed in the blood vessel.

At this time, the number of cells to be analyzed

Figure pat00010
Can count the number of cells to be analyzed in each of a plurality of image signals obtained by imaging blood vessels, and can further improve the reliability by using the calculated values.

8 is a graph showing the number of reference cells and analyzed cells detected from an image obtained according to an embodiment of the present invention according to the flow of time and the number of detected reference cells and the number of cells to be analyzed FIG. 2 is a graph showing an average value obtained by the following equation.

Referring to FIG. 8, the image processing apparatus according to the present embodiment acquires a plurality of image signals every predetermined time after injecting 1 million reference cells and one or more analysis target cells, The number of reference cells to be analyzed and the number of cells to be analyzed were counted to obtain an average value. Thus, the average number of reference cells circulating in the blood continuously and the average number of cells to be analyzed without staying in the organ such as organs were calculated to be 122.8 and 9.7, respectively.

Thereafter, a calibration coefficient (C), which can be converted into the number of circulating cells in the whole blood, is obtained using the average number of reference cells detected.

In this example, the average number of reference cells detected per 30 seconds among the 1 million reference cells injected into blood is 122.8, so that the number of reference cells detected per 30 seconds can be converted into the number of reference cells circulating in the whole blood The calibration coefficient (C) is calculated as 8,143 (= 1,000,000 ÷ 122.8).

The calculation unit 300 calculates the number of cells to be analyzed circulating in the whole blood using the calculated calibration coefficient C and the average value of the number of cells to be analyzed per 30 seconds.

For example, in the present embodiment, the calibration coefficient (C) is 8,143 and the average value of the number of cells analyzed per 30 seconds is 9.7, so that the number of cells to be analyzed circulating in the whole blood is 78,990 (= 8,143 × 9.7).

As described above, the image processing apparatus according to the present embodiment can quantitatively analyze cells circulating in whole blood in a living body by using reference cells continuously circulating without remaining in the body.

9 is a flowchart of an image processing method according to an embodiment of the present invention.

Referring to FIG. 9, the image processing apparatus 100 stores the number of cells to be analyzed and reference cells injected into a living body (S110). The subject cell to be analyzed may be a blood cancer cell present in the bloodstream or a cell expressing a specific protein. In this embodiment, the subject cell to be analyzed may be a colon cancer cell, but the present invention is not limited thereto. Cells to be analyzed may be labeled with a first fluorescent sample. A reference cell is a reference cell that continuously circulates in the bloodstream, and may be, for example, red blood cells or nanoparticles. The reference cell is labeled with a second fluorescent sample and is distinguished from the analyte cell. On the other hand, there may be a plurality of types of cells to be analyzed, and different fluorescent samples may be labeled for different types of cells to be analyzed.

The image processing apparatus 100 captures a specific blood vessel (S120). In this embodiment, the Great Saphenous Vein (GSV) was photographed to fix a large amount of blood and to minimize dynamic noise by tightly fixing blood vessels, but this is not necessarily the case.

The image processing apparatus 100 analyzes the photographed image and acquires the number of reference cells and cells to be analyzed (S130). For example, the image processing apparatus 100 may calculate an average value by counting the number of reference cells and cells to be analyzed at predetermined time intervals after injecting the reference cells and the cells to be analyzed.

The image processing apparatus 100 converts the number of cells to be analyzed circulating in the whole blood using the number of reference cells and the number of cells to be analyzed (S140). In step S110, the number of reference cells labeled with the second fluorescent sample and the average number of reference cells per 30 seconds are used to calculate a calibration coefficient (C) that can be converted into the number of circulating cells in the whole blood ). The calibration coefficient (C) is a ratio of a reference cell passing through a blood vessel among reference cells injected into a living body, and can be defined as shown in Equation (1). The image processing apparatus 100 can convert the number of cells to be analyzed circulating in the whole blood using the calculated correction coefficient C and the average value of the number of cells to be analyzed per 30 seconds.

As described above, the image processing method according to the present embodiment can quantitatively analyze cells circulating in whole blood in a living body by using a reference cell that circulates continuously without remaining in the body.

The embodiments of the present invention described above are not implemented only by the apparatus and method, but may be implemented through a program for realizing the function corresponding to the configuration of the embodiment of the present invention or a recording medium on which the program is recorded.

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, It belongs to the scope of right.

Claims (15)

A photographing unit photographing a blood vessel at a specific position and outputting a video signal, and
An imaging unit that acquires the number of the reference cells and the cells to be analyzed sensed from the image signal and converts the number of cells to be analyzed circulated in the living body using the number of reference cells and the number of cells to be analyzed Device. The method of claim 1,
The operation unit
Calculating a calibration coefficient using the number of reference cells injected into the living body and the number of reference cells sensed in the blood vessel, and calculating a calibration coefficient using the calibration coefficient and the number of cells analyzed in the blood vessel An image processing apparatus for estimating the number of cells to be analyzed. 3. The method of claim 2,
Wherein the calibration coefficient is a ratio of reference cells passing through the blood vessel among the reference cells injected into the living body. 4. The method of claim 3,
The number of reference cells detected in the blood vessel is
Wherein the average value of the number of reference cells calculated in each of the plurality of image signals of the blood vessels. 4. The method of claim 3,
Wherein the calculation unit converts the number of cells to be analyzed circulated in the living body from the number of cells to be analyzed detected in the blood vessel using the calibration coefficient. The method of claim 1,
Wherein the reference cell is a cell that continuously circulates in the living body. The method of claim 1,
Wherein the analyte cell is a cell labeled with a first fluorescent sample and the reference cell is a cell labeled with a second fluorescent sample. The method of claim 1,
The photographing unit
A light irradiation unit for outputting at least one laser light source,
A beam scanning unit for reading the image observed from the blood vessel and generating a two-dimensional array of scanned images,
And an image signal sensing unit for passing the scanning image through a pinhole provided on a confocal plane to generate an image including a fluorescent material excited by the laser light source. An image processing method of an image processing apparatus,
Storing the number of cells to be analyzed and reference cells injected into the living body,
Analyzing an image of a specific blood vessel, obtaining the number of reference cells and the number of cells to be analyzed, which are detected in the blood vessel, and
And converting the number of cells to be analyzed circulating in the living body using the number of reference cells and the number of cells to be analyzed. The method of claim 9,
The step of converting the number of cells to be analyzed circulated in the living body
Calculating a calibration coefficient using the number of reference cells injected into the living body and the number of reference cells sensed in the blood vessel, and
And estimating the number of cells to be analyzed circulated in the living body using the calculated calibration coefficient and the number of cells to be analyzed detected in the blood vessel. 11. The method of claim 10,
Wherein the step of calculating the calibration coefficient is performed by a ratio of the reference cells detected in the blood vessel among the reference cells injected into the living body. 12. The method of claim 11,
The number of reference cells detected in the blood vessel is
Wherein the average value of the number of reference cells calculated in each of the plurality of image signals of the blood vessel is calculated. The method of claim 12,
The step of estimating the number of cells to be analyzed circulated in the living body
And calculating the calibration coefficient by multiplying the calculated calibration coefficient by the number of cells analyzed in the blood vessel. The method of claim 9,
Wherein the reference cell is a cell that continuously circulates in the living body. The method of claim 9,
Wherein the analyte cell is a cell labeled with a first fluorescent sample and the reference cell is a cell labeled with a second fluorescent sample.
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