KR101888926B1 - Method and Apparatus for Analyzing Sperm Cells Using 3D Refractive Index Tomogram - Google Patents

Abstract

A method and an apparatus for analyzing spermatocyte state using three-dimensional refractive index measurement are presented. A method of analyzing spermatocyte state using a three-dimensional refractive index measurement includes optically measuring a three-dimensional refractive index distribution of sperm cells; Extracting a physical quantity of at least one of a shape, a mass, and a concentration of the sperm cell using the measured three-dimensional refractive index distribution; And analyzing the extracted physical quantity of the sperm cells to determine the state of the sperm cells.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method and apparatus for analyzing spermatocytes using 3D refractive index measurement,

The following embodiments are directed to a method and apparatus for analyzing spermatocyte state using three-dimensional refractive index measurement. More particularly, the present invention relates to a method and apparatus for analyzing spermatocyte state using 3D refractive index measurement, more specifically, The present invention relates to a method and apparatus for analyzing spermatocyte state using a three-dimensional refractive index measurement.

Analysis of spermatocyte status is a very important issue in medicine, especially in obstetrics and gynecology. For example, if the shape of the head, trunk, or tail of the sperm is abnormal, it is very difficult for normal correction to occur. Therefore, to determine the presence or absence of such abnormal cells, it is necessary to perform an in vitro fertilization (IVF) to be. In addition, more than 30% of the causes of infertility are in males, and measuring the status and activity of spermatozoa is crucial in determining the cause of these males infertility.

In order to measure the state of sperm cells, the usual method is the morphology and activity analysis using a phase microscope. Phase contrast microscopy (PC) or differential interference contrast microscopy (DIC) are mainly used.

Since it is not possible to use the same method as cell staining because it is a spermatozoon that needs to be injected into the body or to be fertilized, a phase microscope such as PC or DIC can be used to determine the overall contour of the cell, .

However, the problem of the phase microscope was limited to the fact that only 2D image measurement was possible, and only the outline information of the cell could be obtained qualitatively. For example, if an abnormal structure is present on the side of a sperm cell, it can easily be imaged and provide important information for diagnosis, but it is difficult to detect with an existing two-dimensional phase microscope when the abnormal structure is located above or below other structures . In addition, there is a limitation that an examiner should make diagnosis based on experience only based on approximate shape information of a cell.

Thus, there was a fundamental problem that the accurate state of sperm cells could not be grasped. Also, since the spermatocyte state is classified based on the user's experience, there is a problem that the accuracy is low.

1. Kim, K., et al. (2016). "Optical diffraction tomography techniques for the study of cell pathophysiology." arXiv preprint arXiv: 1603.00592. 2. Lee, K., et al. (2013). "Quantitative phase imaging techniques for the study of cell pathophysiology: from principles to applications." Sens cell staining ors 13 (4): 4170-4191. 3. Wolf, E. (1969). "Three-dimensional structure determination of semi-transparent objects from holographic data." Optics Communications 1 (4): 153-156.

Embodiments describe a method and apparatus for analyzing spermatocyte states using a three-dimensional refractive index measurement. More specifically, a three-dimensional refractive index of a cell is optically measured, and a three- Based on the analysis of the physical quantities of the cells.

The examples optically measure the three-dimensional refractive index distribution of living sperm cells without any dyeing or labeling, and analyze the shape, mass, and concentration of sperm cells using the measured three-dimensional refractive index distribution The present invention also provides a method and apparatus for analyzing spermatocytes using a three-dimensional refractive index measurement method for discriminating the abnormality and activity of sperm cells.

The method of analyzing spermatocyte state using three-dimensional refractive index measurement according to an embodiment includes optically measuring a three-dimensional refractive index distribution of sperm cells; Extracting a physical quantity of at least one of a shape, a mass, and a concentration of the sperm cell using the measured three-dimensional refractive index distribution; And analyzing the extracted physical quantities of the sperm cells to determine the state of the sperm cells.

The step of optically measuring the three-dimensional refractive index distribution of the sperm cells comprises the steps of: injecting the light incident from the light source into the sperm cell; Measuring a transmitted light diffracted by the sperm cells using an interferometer to obtain a plurality of two-dimensional holograms; And measuring the three-dimensional refractive index distribution of the sperm cells using the plurality of two-dimensional holograms.

The step of optically measuring the three-dimensional refractive index distribution of the sperm cells comprises rotating the angle of light incident on the sperm cells to measure the plurality of two-dimensional holograms through the interferometer, Can be analyzed through a transmission or diffraction algorithm to measure the three-dimensional refractive index distribution of the sperm cells.

The step of optically measuring the three-dimensional refractive index distribution of the sperm cells may include directly rotating the sperm cells to measure the plurality of two-dimensional holograms through the interferometer, and measuring the two- Diffraction algorithm can be used to measure the three-dimensional refractive index distribution of the sperm cells.

The step of optically measuring the distribution of the three-dimensional refractive index of the sperm cells may include measuring the three-dimensional refractive index distribution of the sperm cells by optical measurement of at least one of optical diffraction tomography and optical projection tomography.

The step of extracting the physical quantity of at least one of the shape, mass, and concentration of the sperm cells using the measured three-dimensional refractive index distribution is characterized in that the sperm cells are separated from the sperm cells Obtaining an outline of the cell; Wherein the spermatocyte morphology information is acquired using a property that the structures of the sperm cells have different refractive index values, and the sperm cell is obtained by using the refractive index value and the refractive index increment value of the protein as a proportionality constant, Calculating a concentration of a localized region of the sample; And calculating the mass of the sperm cells through the calculated concentration and the volume information of the sperm cells.

The step of analyzing the extracted physical quantity of the sperm cells to determine the state of the sperm cells comprises the steps of: analyzing the extracted physical quantity of the sperm cells to determine an abnormal state of the sperm cells; And measuring the activity of the sperm cells by determining the state of the sperm cells at predetermined time intervals.

The apparatus for analyzing spermatocytes according to another embodiment uses a three-dimensional refractive index measurement unit for optically measuring a three-dimensional refractive index distribution of sperm cells. A physical quantity calculating unit for extracting at least one physical quantity of shape, mass, and concentration of the sperm cells using the measured three-dimensional refractive index distribution; And a state determinator for analyzing the extracted physical quantity of the sperm cells to determine the state of the sperm cells.

Wherein the three-dimensional refractive index measuring unit comprises: a light source for causing light to enter the sperm cell; An interferometer for measuring transmitted light diffracted by the sperm cells to obtain a plurality of two-dimensional holograms; And a measuring unit for measuring a three-dimensional refractive index distribution of the sperm cells using the plurality of two-dimensional holograms, wherein the three-dimensional refractive index measuring unit measures the refractive index of the sperm cells using at least one of optical diffraction tomography and optical projection tomography The three-dimensional refractive index distribution of the spermatids can be measured.

The three-dimensional refractive index measuring unit may measure the plurality of two-dimensional holograms through the interferometer by rotating an angle of light incident on the sperm cells, analyzing the plurality of two-dimensional holograms measured through a transmission or diffraction algorithm The three-dimensional refractive index distribution of the spermatids can be measured.

The three-dimensional refractive index measuring unit may measure the plurality of two-dimensional holograms through the interferometer by directly rotating the sperm cells, analyzing the plurality of two-dimensional holograms measured through a transmission or diffraction algorithm, Dimensional refractive index distribution can be measured.

Wherein the physical quantity calculation unit comprises: a shape extracting unit that obtains an outline of the sperm cells using characteristics of the sperm cells different from each other in refractive index values; Wherein the spermatocyte morphology information is acquired using a property that the structures of the sperm cells have different refractive index values, and the sperm cell is obtained by using the refractive index value and the refractive index increment value of the protein as a proportionality constant, A concentration extracting unit for calculating a concentration of a local region of the sample; And a mass extractor for calculating the mass of the sperm cells through the calculated concentration and the volume information of the sperm cells.

Wherein the state determiner comprises: an abnormal state determiner for analyzing the extracted physical quantity of the sperm cells to determine an abnormal state of the sperm cells; And an activity measuring unit for measuring the activity of the sperm cells by observing the state of the sperm cells at predetermined time intervals.

According to still another embodiment of the present invention, there is provided an apparatus for analyzing spermatocytic cells using three-dimensional refractive index measurement, comprising: a light source for introducing light into cells; An interferometer for measuring transmitted light diffracted in the cell to obtain a plurality of two-dimensional holograms; And a measuring unit for measuring a three-dimensional refractive index distribution of the cell using the plurality of two-dimensional holograms, wherein the three-dimensional refractive index distribution of the cell is measured through optical measurement of at least one of optical diffraction tomography and optical projection tomography And determines the state of the cell using the measured three-dimensional refractive index distribution.

A physical quantity calculation unit for extracting at least one physical quantity of the shape, mass, and concentration of the cell using the measured three-dimensional refractive index distribution of the cell; And a state determination unit for analyzing the extracted physical quantity of the cell to determine the state of the cell.

According to the embodiments, the three-dimensional refractive index distribution of living sperm cells is optically measured without any dyeing or labeling, and the shape, mass, and concentration of sperm cells are measured using the measured three-dimensional refractive index distribution Based on the analysis, it is possible to provide a method and apparatus for analyzing spermatocyte state using a three-dimensional refractive index measurement for discriminating an abnormality and activity of spermatids.

According to the embodiments, when the state and activity of sperm cells are measured, the three-dimensional refractive index distribution is measured to overcome the limitation of the two-dimensional image of the conventional phase microscope, and the information (mass, A method of analyzing spermatocyte state and an apparatus using the three-dimensional refractive index measurement capable of quantitatively measuring the concentration of the spermatozoa of the present invention can be provided.

1 is a view for explaining a sperm cell status analysis method using a three-dimensional refractive index measurement according to an embodiment.
FIG. 2 is a view for explaining a sperm cell state analyzing apparatus using a three-dimensional refractive index measurement according to an embodiment.
3A is a view for explaining a method of measuring a three-dimensional refractive index of a cell using an incident light rotation method according to an embodiment.
FIG. 3B is a view for explaining a method of measuring the three-dimensional refractive index of cells using the cell rotation method according to one embodiment.
FIGS. 4 to 6 are flowcharts for explaining a sperm cell state analysis method using a three-dimensional refractive index measurement according to an embodiment.

Hereinafter, embodiments will be described with reference to the accompanying drawings. However, the embodiments described may be modified in various other forms, and the scope of the present invention is not limited by the embodiments described below. In addition, various embodiments are provided to more fully describe the present invention to those skilled in the art. The shape and size of elements in the drawings may be exaggerated for clarity.

The following examples provide a method for analyzing the state of living sperm cells without additional steps such as staining or labeling. More specifically, the embodiments are capable of optically measuring the three-dimensional refractive index distribution of a cell and determining the abnormality and activity of the sperm cell based on the shape / mass / concentration analysis of the cell.

For example, when a laser beam is irradiated from a light source to a cell (sample), a plurality of 2D images can be acquired while the laser beam is rotating, and a 3D image can be acquired by obtaining a 3D absorption degree by performing tomograms through analysis . Thus, three-dimensional images can be obtained without staining the cells. In the case of confirming 3-dimensional images of cells, it is necessary to stain the cells, but it is difficult to inject into the body after staining of the cells, which makes it difficult to apply to various fields.

1 is a view for explaining a sperm cell status analysis method using a three-dimensional refractive index measurement according to an embodiment.

1A, when measuring the state and activity of sperm cells, conventional phase microscopes acquire two-dimensional images, and it is impossible to quantitatively measure information (mass, concentration of local region) of sperm cells.

Referring to FIG. 1B, when the state and activity of sperm cells are measured, a three-dimensional refractive index distribution is measured to overcome the limitation of a two-dimensional image of a conventional phase microscope, and information Concentration of the region) can be measured quantitatively.

The present embodiments provide a method of measuring the state and activity of a cell using the measurement of three-dimensional refractive index of living sperm cells without using dyeing or marking.

The intracellular 3D refractive index distribution is closely related to the composition and morphology of subcellular organelles. In addition, the refractive index value itself is proportional to the concentration of protein, which is a major component in the cell. Thus, measuring the three-dimensional refractive index information of a cell reflects not only morphological characteristics of cells and intracellular organelles but also biochemical characteristics information.

Accordingly, when the three-dimensional refractive index distribution is used, the shape and state of the cell can be distinguished by using the specific morphological and biochemical characteristics of the cell type.

The shape, mass, and concentration of spermatids can be extracted based on the measurement results of the three-dimensional refractive index distribution of sperm cells. Specifically, the shape of a cell can be obtained by using the difference in the refractive index between the cell and the culture medium.

In addition, the structure of sperm cells (eg, acrosome, nucleus, centriole, mitochondria, axis filament, etc.) have different refractive index values, The shape information is also obtainable.

The local concentration of the cells can be calculated using the measured refractive index value and the refractive index increment value of a protein that is already known. The concentration of the protein solution is directly proportional to the refractive index, where the proportion constant is the refractive index increment. Using this, it is possible to measure the concentration of the cellular local region from the measured refractive index value.

The mass of the cell can also be easily calculated from the measured concentration and the volume information of the cell.

Thus, the three dimensional refractive index distribution of the sperm cells can be measured, and the shape, mass, and concentration of the spermatids can be extracted based on the measured values, and the state and activity of the spermatids can be measured based on the measurements.

Hereinafter, a method and apparatus for analyzing spermatocyte state using three-dimensional refractive index measurement will be described in more detail with an example.

FIG. 2 is a view for explaining a sperm cell state analyzing apparatus using a three-dimensional refractive index measurement according to an embodiment.

Referring to FIG. 2, an apparatus 200 for analyzing sperm cell state using three-dimensional refractive index measurement according to an embodiment includes a three-dimensional refractive index measuring unit 210, a physical quantity calculating unit 220, and a state determining unit 230 .

Here, the three-dimensional refractive index measuring unit 210 may include a light source, an interferometer, and a measuring unit. The physical quantity calculating unit 220 may include a shape extracting unit, a concentration extracting unit, and a mass extracting unit. The state determining unit 230 may include an abnormality determining unit and an activity measuring unit.

Hereinafter, spermatids in the cell will be described more specifically as an example.

The three-dimensional refractive index measuring unit 210 can optically measure the three-dimensional refractive index distribution of sperm cells.

For example, the three-dimensional refractive index measurement unit 210 may be an optical system including a light source and a camera, and may have various shapes such as a reflection type and a transmission type.

The three-dimensional refractive index measurement unit 210 may include a light source, an interferometer, and a measurement unit.

The light source can cause light to enter the cell. That is, the light source can irradiate sperm cells with light. For example, a laser can be used as a light source, and a light source can irradiate a laser beam onto a sample such as sperm cells to be measured.

Here, the cell may be represented by a sample representing an object to be measured, and it may be a bacterium or a microorganism as well as a cell, and may be an object including a cell or the like. For example, the cell may be a sperm cell or an object containing sperm cells.

The light source used may utilize a single wavelength laser. In addition, the light source can utilize a larger amount of information to differentiate cells by measuring the refractive index of three dimensions at each wavelength using a multi-wavelength laser.

The interferometer can acquire a plurality of two-dimensional holograms by measuring the transmitted light diffracted by the sperm cells after the light incident from the light source is incident on the cells.

Here, the interferometer is a measuring device using the interference phenomenon of light, which divides the light emitted from the same light source into two or more beams to make a difference in the propagation path, and then observes the interference phenomenon occurring when the light meets again.

The measuring unit can measure the three-dimensional refractive index distribution of sperm cells using a plurality of two-dimensional holograms obtained from the interferometer. For example, a camera, which is a photographing apparatus for photographing an image, may be used as the measuring unit.

The three-dimensional refractive index measuring unit 210 can measure the three-dimensional refractive index distribution of the spermatids through optical measurement of at least one of optical diffraction tomography and optical projection tomography.

The three-dimensional refractive index measuring unit 210 measures a plurality of two-dimensional holograms measured in the interferometer by rotating the angle of light incident on the sperm cells, analyzes the measured two-dimensional holograms through a transmission or diffraction algorithm, The three-dimensional refractive index distribution of the cell can be measured.

For example, using a wavefront controller to measure the angles of the sperm cells and measure the two-dimensional holograms, and analyze the measured two-dimensional hologram information through transmission or diffraction algorithms, can do. In particular, a high-speed three-dimensional optical tomography can be performed using a digital micromirror device, which is one of wavefront controllers.

The three-dimensional refractive index measuring unit 210 measures a number of two-dimensional holograms measured in the interferometer by directly rotating the sperm cells, analyzes the measured two-dimensional holograms through a transmission or diffraction algorithm, Dimensional refractive index distribution can be measured.

On the other hand, the method of measuring spermatids is a method in which sperm cells are placed at low concentration on an in vitro slide glass, a method in which spermatids are singly or in a high concentration on an in vitro slide glass, Multi-layered forms, tissue slides in which biopsy slides are cut to a thickness of between 5 micrometers and 50 micrometers, and microfluidic tubes for high-throughput screening in vitro (microfluidic channel).

The physical quantity calculation unit 220 can extract at least one physical quantity of the sperm cell shape, mass, and concentration using the three-dimensional refractive index distribution measured by the three-dimensional refractive index measurement unit 210.

The physical quantity calculation unit 220 may include a shape extracting unit, a concentration extracting unit, and a mass extracting unit.

The shape extracting unit can obtain the outline of sperm cells using characteristics of sperm cells and culture fluid having different refractive index values.

The concentration extractor obtains the shape information of the sperm cells using sperm cell structures having different refractive index values and uses the refractive index value and the refractive index increment value of the protein as a proportional constant to calculate the sperm cell The concentration of the local region can be calculated.

The mass extractor can calculate the mass of the spermatids through the calculated concentration and volume information of spermatids.

The state determiner 230 can determine the state of the sperm cells by analyzing the extracted physical quantity of sperm cells.

The state determiner 230 may include an abnormal state determiner and an activity measurer.

The anomalous state discriminator can discriminate an abnormal state of spermatids by analyzing extracted physical quantities of spermatids.

The activity measuring unit can measure the activity of the sperm cells by grasping the state of the sperm cells at predetermined time intervals.

According to the embodiments, the three-dimensional refractive index distribution of living sperm cells is optically measured without any dyeing or labeling, and the shape, mass, and concentration of sperm cells are measured using the measured three-dimensional refractive index distribution Based on the analysis, it is possible to provide a method and apparatus for analyzing spermatocyte state using a three-dimensional refractive index measurement for discriminating an abnormality and activity of spermatids.

Hereinafter, a method of measuring the three-dimensional refractive index distribution of a cell through optical measurement will be described in more detail.

3A is a view for explaining a method of measuring a three-dimensional refractive index of a cell using an incident light rotation method according to an embodiment.

All objects have a refractive index distribution. The refractive index is the intrinsic optical quantity of the material itself, which describes how much the velocity slows down as the light passes through it. Optical diffraction tomography or optical projection tomography (tomographic phase microscopy, 3D digital holographic microscopy) can be used to measure the three-dimensional refractive index of cells [Non-Patent Documents 1 and 2].

Referring to FIG. 3A, the optical diffraction tomography and the optical projection tomography can use the same optical implementation (Non-Patent Document 3). The light emitted from the coherent light source 310 is incident on the cell 300 and the transmitted light diffracted by the cell 300 can be measured using the interferometer 320. Thereafter, the measuring unit can measure the three-dimensional refractive index distribution 340 of the cell 300 by analyzing the measured two-dimensional hologram information through the transmission or diffraction algorithm 330.

At this time, the three-dimensional refractive index profile 340 of the cell 300 can be measured using several two-dimensional holograms measured while rotating (scanning) the angle of light (incident light) incident on the cell 300. Here, the difference between the optical diffraction tomography and the optical projection tomography is the restoration algorithm which considers the diffraction of light in the specimen.

On the other hand, the method of measuring the cell 300 may include a method in which cells are placed at low concentration on an in vitro slide glass, a method in which the cells are singly or in a high concentration on an in vitro slide glass, Multi-layered forms, tissue slides in which biopsy slides are cut to a thickness of between 5 micrometers and 50 micrometers, and microfluidic tubes for high-throughput screening in vitro (microfluidic channel).

And the light source 310 used may utilize a single wavelength laser. Further, by measuring the three-dimensional refractive index (340) at each wavelength by using the multiple wavelength laser as the light source 310, a larger amount of information can be utilized for the classification of the cells 300.

FIG. 3B is a view for explaining a method of measuring the three-dimensional refractive index of cells using the cell rotation method according to one embodiment.

Referring to FIG. 3B, in the method of measuring the three-dimensional refractive index of the cell 300 using the incident light rotation method according to the embodiment described with reference to FIG. 3A, instead of rotating the incident light, (340).

Using the method described above, the 3D refractive index image of living sperm cells can be measured, and the state and activity of sperm cells can be measured using the measured three-dimensional refractive index distribution.

FIGS. 4 to 6 are flowcharts for explaining a sperm cell state analysis method using a three-dimensional refractive index measurement according to an embodiment.

Referring to FIG. 4, a method for analyzing spermatocyte state using three-dimensional refractive index measurement according to an exemplary embodiment of the present invention includes a step 430 of optically measuring a three-dimensional refractive index distribution of sperm cells, Extracting a physical quantity of at least one of the shape, mass, and concentration of the cell (440), and analyzing the extracted physical quantity of the sperm cell to determine the state of the sperm cell (450).

Meanwhile, the method of analyzing spermatocytes using three-dimensional refractive index measurement according to an embodiment may further include a step 410 of loading a sperm cell into a specimen part of an imaging apparatus and a step 420 of confirming a specimen to be measured .

5, the step of optically measuring the three-dimensional refractive index profile of the sperm cells comprises the steps of (431) entering light into the sperm cells from a light source, measuring the transmitted light diffracted by the sperm cells using an interferometer A step 432 of obtaining a plurality of two-dimensional holograms, and a step 433 of measuring a three-dimensional refractive index distribution of the sperm cells using the plurality of two-dimensional holograms.

Referring to FIG. 6, the step of extracting at least one of the shape, mass, and concentration of the sperm cells using the measured three-dimensional refractive index distribution may include: (441) of obtaining an outline of the sperm cell using the characteristic of the sperm cell, and acquiring the shape information of the sperm cell using the property that the structure of the sperm cell has different refractive index values, Calculating a concentration of a local region of the sperm cell using a refractive index increment value as a proportionality constant, and calculating the concentration of the sperm cell by using the calculated concentration and the volume information of the sperm cell (Step 443). ≪ / RTI >

According to the embodiments, when the state and activity of sperm cells are measured, the three-dimensional refractive index distribution is measured to overcome the limitation of the two-dimensional image of the conventional phase microscope, and the information (mass, A method of analyzing spermatocyte state and an apparatus using the three-dimensional refractive index measurement capable of quantitatively measuring the concentration of the spermatozoa of the present invention can be provided.

Hereinafter, a method for analyzing sperm cell state using three-dimensional refractive index measurement according to one embodiment will be described in more detail with reference to one embodiment.

The sperm cell state analysis method using the three-dimensional refractive index measurement according to one embodiment can be more specifically explained using the sperm cell state analyzer using the three-dimensional refractive index measurement according to the embodiment described in FIG. Here, the sperm cell state analyzer 200 using the three-dimensional refractive index measurement according to an embodiment includes a three-dimensional refractive index measuring unit 210, a physical quantity calculating unit 220, and a state determining unit 230 have.

First, in the sperm cell state analysis method using the three-dimensional refractive index measurement according to an embodiment, in step 410, the sperm cell state analyzer 200 using the three-dimensional refractive index measurement measures sperm cells to be measured, Can be loaded on the part.

Then, in step 420, the sperm cell state analyzer 200 using the three-dimensional refractive index measurement can confirm the specimen to be measured.

In step 430, the three-dimensional refractive index measurement unit 210 can optically measure the three-dimensional refractive index distribution of the cell.

More specifically, in order to optically measure the three-dimensional refractive index profile of the cell, the three-dimensional refractive index measurement unit 210 may cause the light source to enter the cell in step 431. Then, in step 432, the interferometer can acquire a plurality of two-dimensional holograms by measuring the transmitted light diffracted by the cells after the light incident from the light source is incident on the cells. Thereafter, in step 433, the measuring unit can measure the three-dimensional refractive index distribution of the cell using a plurality of two-dimensional holograms obtained in the interferometer.

The three-dimensional refractive index measuring unit 210 can measure the three-dimensional refractive index distribution of the cell through optical measurement of at least one of optical diffraction tomography and optical projection tomography.

The three-dimensional refractive index measuring unit 210 may measure the three-dimensional refractive index distribution of the cell using a plurality of two-dimensional holograms measured in the interferometer by rotating the angle of light incident on the cell.

The three-dimensional refractive index measuring unit 210 may also measure the three-dimensional refractive index distribution of the cells using a plurality of two-dimensional holograms measured in the interferometer by directly rotating the cells.

In step 440, the physical quantity calculation unit 220 can extract the physical quantity of the shape, mass, and concentration of sperm cells using the measured three-dimensional refractive index distribution.

More specifically, in step 441, the physical quantity calculation unit 220 can obtain the outline of the sperm cells using characteristics of the sperm cells and the culture fluid having different refractive index values.

In step 442, the physical quantity calculation unit 220 determines the structure of the sperm cell (for example, acrosome, nucleus, centriole, mitochondria, axis filament, etc.) The spermatocyte shape (or the structure of the intracellular organelles) was obtained by using characteristics having different refractive index values, and the spermatocyte counts were obtained by using the refractive index value and the refractive index increment value of the protein as a proportional constant Can be calculated.

Here, the concentration of the protein solution is directly proportional to the refractive index, where the proportion constant is the refractive index increment. Using this, it is possible to measure the concentration of the cellular local region from the measured refractive index value.

In step 443, the physical quantity calculation unit 220 can calculate the sperm cell mass through the calculated concentration and the volume information of the sperm cell.

Thus, the state of sperm cells can be grasped through various physical quantities of acquired sperm cells.

In operation 450, the state determiner 230 may determine the state of the sperm cells by analyzing the extracted physical quantity of sperm cells.

More specifically, the state determiner 230 may analyze the extracted physical quantity of the sperm cells to determine whether the sperm cells are abnormal.

The state determiner 230 can measure the activity of the sperm cells by grasping the state of the sperm cells at predetermined time intervals.

In particular, in step 460, the state determining unit 230 may terminate the inspection according to whether the inspection is repeated or confirm the measurement target specimen again (420). Thus, the sperm cell activity can be measured at each time point, so that the activity of the sperm can also be measured.

Therefore, when measuring the state and activity of sperm cells, a three-dimensional refractive index distribution and quantitative cell information (mass, concentration of local region) can be utilized. This technology can be widely used in medicine, especially in obstetrics and gynecology. For example, when determining the state of spermatids for infertility testing or in vitro fertilization, we need to overcome the limitations of 2D images of conventional phase microscopy and to use the cell information (mass, concentration of local region) It can be measured quantitatively.

The sperm cell state analyzing apparatus using the three-dimensional refractive index measurement according to another embodiment may include a light source, an interferometer, and a measuring unit. Each constitution of the sperm cell state analyzing apparatus using the three-dimensional refractive index measurement according to this other embodiment is identical to that of the sperm cell state analyzing apparatus using the three-dimensional refractive index measurement according to the embodiment described in FIGS. 2 and 3 I will explain.

The light source can cause light to enter the cell. That is, the light source can irradiate cells with light. For example, a laser can be used as a light source, and a light source can irradiate a laser beam onto a sample such as a cell to be measured.

The light source used may utilize a single wavelength laser. In addition, the light source can utilize a larger amount of information to differentiate cells by measuring the refractive index of three dimensions at each wavelength using a multi-wavelength laser.

The interferometer can acquire a plurality of two-dimensional holograms by measuring the transmitted light diffracted by the cell after the light incident from the light source is incident on the cell.

Here, the interferometer is a measuring device using the interference phenomenon of light, which divides the light emitted from the same light source into two or more beams to make a difference in the propagation path, and then observes the interference phenomenon occurring when the light meets again.

The measuring unit can measure the three-dimensional refractive index distribution of the cell using a plurality of two-dimensional holograms obtained from the interferometer. For example, a camera, which is a photographing apparatus for photographing an image, may be used as the measuring unit.

For example, by measuring the number of two-dimensional holograms measured by an interferometer by rotating the angle of light incident on the cell, and analyzing the measured two-dimensional holograms through a transmission or diffraction algorithm, Can be measured.

As another example, a plurality of two-dimensional holograms measured by an interferometer may be measured by directly rotating the cells, and a plurality of measured two-dimensional holograms may be analyzed through a transmission or diffraction algorithm to measure a three-dimensional refractive index distribution of the cells.

The sperm cell state analyzing apparatus using the three-dimensional refractive index measurement according to this another embodiment measures the three-dimensional refractive index distribution of cells through optical measurement of at least one of optical diffraction tomography and optical projection tomography, The refractive index distribution can be used to determine the state of the cell.

The sperm cell state analyzing apparatus using the three-dimensional refractive index measurement according to another embodiment may further include a physical quantity calculating unit and a state determining unit.

The physical quantity calculation unit can extract at least one physical quantity of the shape, mass, and concentration of the cell using the measured three-dimensional refractive index distribution.

The state determiner can determine the state of the cell by analyzing the extracted physical quantity of the cell.

According to the embodiments, the three-dimensional refractive index distribution of living cells is optically measured and the abnormality of the cell is determined based on the shape, mass, and concentration of the cell using the measured three-dimensional refractive index distribution .

Further, according to the embodiments, the type of the cell can be distinguished without using dyeing or labeling by measuring the distribution of the refractive index of the third month. This can be widely used in the field of cell biology, particularly in the field of cytometry.

For example, since a stem cell can be identified by a non-labeled non-staining method, it can be injected into the body immediately. As another example, when the present invention is applied to the field of immunotherapy in which a specific type of leukocyte extracted from blood of a cancer patient is selectively cultured and re-infused, the conventional magnetic cell-based cell sorting (MACS, magnetic-assisted cell cytometry can be effectively solved.

It is also applicable to medical infections in medicine. For example, by making it possible to quickly and accurately distinguish between bacterial species, a medical prescription, such as antibiotics, can be made quickly and effectively for infected patients. Another example is the ability to quickly and accurately screen toxic fungi (eg, anthrax) that can be dedicated to weapons, and thus can be used in counter-terrorism applications.

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, controller, arithmetic logic unit (ALU), digital signal processor, microcomputer, 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 apparatus 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 embodyed temporarily. 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 (15)

Optically measuring the three-dimensional refractive index profile of un-stained unlabeled sperm cells;
Extracting physical quantities of shape, mass, and concentration of the sperm cells using the measured three-dimensional refractive index profile; And
Determining the state of the sperm cells by analyzing the physical quantity extracted from the sperm cells based on morphological and chemical characteristics of the sperm cells
Lt; / RTI >
The step of optically measuring the three-dimensional refractive index distribution of the sperm cells comprises:
Dimensional hologram is measured through an interferometer by rotating an angle of light incident on the sperm cell or directly rotating the sperm cell to measure the plurality of two-dimensional holograms through the interferometer, Dimensional hologram is analyzed to measure the three-dimensional refractive index distribution of the sperm cells,
The step of optically measuring the three-dimensional refractive index distribution of the sperm cells comprises:
Introducing light incident from a light source into the sperm cell;
Measuring a transmitted light diffracted by the sperm cells using an interferometer to obtain a plurality of two-dimensional holograms; And
Measuring a three-dimensional refractive index distribution of the sperm cells using the plurality of two-dimensional holograms
Lt; / RTI >
Dimensional hologram through the interferometer by rotating an angle of light incident on the sperm cell using a digital micromirror device for ultrafast three-dimensional optical tomography, measuring the plurality of two-dimensional holograms through the interferometer, Dimensional hologram of the sperm cells is analyzed through a transmission or diffraction algorithm to measure the three-dimensional refractive index distribution of the sperm cells,
The step of extracting physical quantities for shape, mass, and concentration of the sperm cells using the measured three-dimensional refractive index distribution may include:
Obtaining an outline of the sperm cells using different characteristics of the sperm cells and the culture solution;
At least two or more of the acrosome, nucleus, centriole, mitochondria, and axis filament of the structure of the spermatids have different refractive index values, The intracellular shape information is obtained and the concentration of the local region of the spermatids is calculated using the refractive index value and the refractive index increment value of the protein as a proportional constant through the characteristic that the concentration of the protein solution is directly proportional to the refractive index ; And
Calculating the mass of the sperm cells through the calculated concentration and the volume information of the sperm cell
A method for analyzing sperm cell status using three-dimensional refractive index measurement. delete delete delete The method according to claim 1,
The step of optically measuring the three-dimensional refractive index distribution of the sperm cells comprises:
Measuring the three-dimensional refractive index distribution of the sperm cells through optical measurement of at least one of optical diffraction tomography and optical projection tomography
A method for analyzing spermatocytes using a three-dimensional refractive index measurement. delete The method according to claim 1,
The step of analyzing the extracted physical quantity of the sperm cells to determine the state of the sperm cells comprises the steps of:
Analyzing the extracted physical quantity of the sperm cells to determine an abnormal state of the sperm cells; And
Determining the state of the sperm cells at predetermined time intervals and measuring the activity of the sperm cells
A method for analyzing sperm cell status using three-dimensional refractive index measurement. A three-dimensional refractive index measuring unit for optically measuring a three-dimensional refractive index distribution of a non-stained non-labeled sperm cell;
A physical quantity calculating unit for extracting physical quantities for shape, mass, and concentration of the sperm cells using the measured three-dimensional refractive index distribution; And
A state discriminating unit for analyzing the extracted physical quantity of the sperm cells based on morphological characteristics and chemical characteristics of the sperm cells to determine the state of the sperm cells
Lt; / RTI >
Wherein the three-dimensional refractive index measuring unit comprises:
Dimensional hologram is measured through an interferometer by rotating the angle of light incident on the sperm cell or directly rotating the sperm cell to measure the plurality of two-dimensional holograms through the interferometer, Dimensional hologram is analyzed to measure the three-dimensional refractive index distribution of the sperm cells,
Wherein the three-dimensional refractive index measuring unit comprises:
A light source for introducing light into sperm cells;
An interferometer for measuring transmitted light diffracted by the sperm cells to obtain a plurality of two-dimensional holograms; And
A measurement unit for measuring a three-dimensional refractive index distribution of the sperm cells using the plurality of two-dimensional holograms,
Lt; / RTI >
Dimensional hologram through the interferometer by rotating an angle of light incident on the sperm cell using a digital micromirror device for ultrafast three-dimensional optical tomography, measuring the plurality of two-dimensional holograms through the interferometer, Dimensional hologram of the sperm cells is analyzed through a transmission or diffraction algorithm to measure the three-dimensional refractive index distribution of the sperm cells,
The physical quantity calculation unit may calculate,
A shape extracting unit for acquiring an outline of the sperm cells using characteristics of the sperm cells and the culture solution having different refractive index values;
At least two or more of the acrosome, nucleus, centriole, mitochondria, and axis filament of the structure of the spermatids have different refractive index values, The intracellular shape information is obtained and the concentration of the local region of the spermatids is calculated using the refractive index value and the refractive index increment value of the protein as a proportional constant through the characteristic that the concentration of the protein solution is directly proportional to the refractive index ; And
A mass extracting unit for calculating the mass of the sperm cells through the calculated concentration and the volume information of the sperm cells,
And measuring the refractive index of the sperm cell using the three-dimensional refractive index measurement. 9. The method of claim 8,
Wherein the three-dimensional refractive index measuring unit comprises:
Measuring the three-dimensional refractive index distribution of the sperm cells through optical measurement of at least one of optical diffraction tomography and optical projection tomography
And measuring the refractive index of the spermatids. delete delete delete 9. The method of claim 8,
Wherein the state determiner comprises:
An abnormal state discrimination unit for analyzing the extracted physical quantity of the sperm cells to determine an abnormal state of the sperm cells; And
An activity measurement unit for measuring the activity of the sperm cells by grasping the state of the sperm cells at predetermined time intervals,
And measuring the refractive index of the sperm cell using the three-dimensional refractive index measurement. delete delete

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