KR100963239B1 - Medical polarization microscope system and Method for measuring double refraction by polarization modulation - Google Patents

Abstract

The present invention relates to a medical polarization microscope system and a method for measuring birefringence characteristics by polarization adjustment. More specifically, the birefringence characteristics of living cells or tissues are measured by adjusting polarization by adjusting one of the two polarizers constituting the polarizer or controlling the phase delay value of the liquid crystal variable phase delayer with an electrical signal. By measuring the birefringence characteristics of normal and cancer cells through the measurement of the cancer cells, it is possible to diagnose cancer cells, and also to measure the birefringence characteristics by medical polarization microscopy system and polarization adjustment to identify the metastasis and differentiation characteristics of the cells It is about a method.

To this end, the present invention, the light generating unit for generating light; A lens unit converting the generated light into parallel light; A polarizer which adjusts a polarization state of light incident from the lens unit and includes a first polarizer and a second polarizer; A phase delay unit provided between the first polarizer and the second polarizer; A measuring unit configured to pass the light passing through the first polarizer through a sample to be measured; And a light detector for detecting the polarization state of the light passing through the sample as the light intensity.

 Polarization microscope, polarizer, liquid crystal variable phase retarder, birefringence, polarization adjustment

Description

Medical polarization microscope system and method for measuring double refraction by polarization modulation}

The present invention relates to a medical polarization microscope system and a method for measuring birefringence characteristics by polarization adjustment. More specifically, the birefringence characteristics of living cells or tissues are measured by adjusting polarization by adjusting one of the two polarizers constituting the polarizer or controlling the phase delay value of the liquid crystal variable phase delayer with an electrical signal. By measuring the birefringence characteristics of normal and cancer cells through the measurement of the cancer cells, it is possible to diagnose cancer cells, and also to measure the birefringence characteristics by medical polarization microscopy system and polarization adjustment to identify the metastasis and differentiation characteristics of the cells It is about a method.

Microscope systems currently used for medical purposes, such as diagnosing cancer cells and cancer tissues, are mostly used to image and display characteristics of living cells and tissues so that specific living cells and tissues may be normal. To this end, generally, a method of attaching a fluorescent tag or performing dyeing using a specific living cell or tissue as a target has been used.

According to this method, the characteristics of living cells and tissues can be imaged only through a complicated pretreatment process, and thus there is a problem that the measurement takes much time.

On the other hand, it has recently been found that living cells and tissues have birefringent properties. Accordingly, studies for using the birefringent properties of the living cells and tissues in the diagnosis of cancer cells and cancer tissues are being actively conducted.

For example, a study has been introduced on a polarizing microscope that measures the birefringence characteristics of living cells and tissues by controlling the polarization state of the light source by a magnetic field (Modulated Polarization Microscopy, J. R. Kuhn, Biophysical Journal, 2001). The method uses a magnetic field to control the polarization state of light, but a large magnetic field complicates the system, and by adjusting the polarization state of the light to a magnetic field, energy consumption and heat generation are large, Due to the difficulty in accurate control, there is a problem that the accuracy is lowered.

Therefore, there is a need for the development of a medical polarization microscope system and a birefringence characteristic measurement method that can adjust the polarization state with a simpler system and method, while reducing energy consumption and improving the accuracy of the birefringence characteristic measurement.

The present invention has been made to solve the above problems, in particular to measure the birefringence characteristics of living cells or tissues, and to determine the birefringence characteristics of normal cells and cancer cells through the measurement of such birefringence characteristics to diagnose cancer cells It is an object of the present invention to provide a method for measuring birefringence properties by medical polarization microscopy system and polarization adjustment which can enable and identify the metastasis and differentiation characteristics of cells.

Medical polarization microscope system according to an aspect of the present invention devised to achieve the above object is a light generating unit for generating light; A lens unit converting the generated light into parallel light; A polarizer which adjusts a polarization state of light incident from the lens unit and includes a first polarizer and a second polarizer; A phase delay unit provided between the first polarizer and the second polarizer; A measuring unit configured to pass the light passing through the first polarizer through a sample to be measured; And a light detector for detecting the polarization state of the light passing through the sample as the light intensity.

Medical polarization microscope system according to another aspect of the present invention includes a light generating unit for generating light; A lens unit converting the generated light into parallel light; A polarizer including a polarizer generating polarized light by receiving parallel light from the lens unit, and a liquid crystal variable phase retarder for adjusting the polarized light by controlling a phase delay of the polarized light by an electric signal; A measuring unit which passes the light passing through the polarizing unit to the sample to be measured; And a light detector for detecting the polarization state of the light passing through the sample as the light intensity.

The method for measuring birefringence characteristics by polarization adjustment according to the present invention includes the steps of (a) generating light; (b) converting the generated light into parallel light; (c) adjusting the polarization state of the sample whose birefringence characteristic is to be measured; And (d) collecting the light intensity according to the polarization adjustment to measure the birefringence characteristics of the sample including the direction of the main axis and the phase delay value.

According to the present invention, by adjusting one of the two polarizers constituting the polarizer, the birefringence characteristics of living cells or tissues are measured without using cell staining or fluorescent markers, and the birefringence of normal cells and cancer cells is measured by measuring the birefringence characteristics. By identifying the difference in characteristics enables the diagnosis of cancer cells, and also has the effect of identifying the metastasis and differentiation characteristics of the cells.

In addition, according to the present invention, by controlling the phase delay value of the liquid crystal variable phase delay unit with an electrical signal, polarization can be adjusted at a high speed, and thus the birefringence characteristic can be measured in real time.

Hereinafter, exemplary embodiments of the present invention will be described in detail with reference to the accompanying drawings. First, in adding reference numerals to the components of each drawing, it should be noted that the same reference numerals are assigned to the same components as much as possible, even if shown on different drawings. In the following description of the present invention, a detailed description of known functions and configurations incorporated herein will be omitted when it may make the subject matter of the present invention rather unclear. In addition, the following will describe a preferred embodiment of the present invention, but the technical idea of the present invention is not limited thereto and may be variously modified and modified by those skilled in the art.

1 is a block diagram of a medical polarization microscope system according to an embodiment of the present invention.

In the medical polarization microscope system according to the exemplary embodiment of the present invention, referring to FIG. 1, the light generating unit 110, the lens unit 120, the polarizing units 142 and 154, the phase delay unit 144, and the measuring unit And a light detector 160 and a light detector 160. In addition, a first light control unit 115 and a second light control unit 130 are provided between the light generator 110 and the lens unit 120 and between the lens unit 120 and the polarizer 142, respectively. .

The light generator 110 is a light source that generates light for measuring birefringence characteristics of a sample such as living cells and tissues. It is preferable that a laser is used as the light source of the light generator 110, and more preferably, a helium neon or an ar-ion laser may be used. Since a helium neon laser or an argon-ion laser has a very narrow spectrum and can be regarded as a light source of almost short wavelength, such short wavelength light may exhibit specific birefringence characteristics when transmitted through a sample. However, the type of the light source is not limited thereto.

The first light controller 115 changes the advancing direction of the light generated by the light generator 110. A reflector may be used as the first light control unit 115. The light generated by the light generating unit 110 is reflected by the first light adjusting unit 115 and is changed into the lens unit 120 after the traveling direction is changed.

The lens unit 120 converts the light generated by the light generator 110 into parallel light. The lens unit 120 includes a diffuser 122, a parallel light lens 124, and an aperture 126.

The optical diffuser 122 completely scatters and emits the light reflected by the first light control unit 115. At this time, the diffuser 122 serves to remove various types of diffraction patterns due to the spatial coherence of the laser appearing in the image. The diffuser 122 may be rotated at high speed to obtain a temporally uniform image.

A plano-convex lens 124 converts the light passing through the diffuser 122 into parallel light.

The aperture 126 adjusts the amount of light passing through the parallel light lens 124 so that an appropriate amount of light may be incident on the polarizer 142.

The second light control unit 130 serves to change the traveling direction of the light passing through the lens unit 120. As the second light control unit 130, a reflector may be used as the first light control unit 115. The light passing through the lens unit 120 is reflected by the second light control unit 130, and the traveling direction is changed, and then the light is incident to the polarizer 142.

The polarizers 142 and 154 adjust the polarization state of light incident from the lens unit 120 and include a first polarizer 142 and a second polarizer 154. The first polarizer 142 is located between the incident light side, that is, between the second light control unit 130 and the phase delay unit 144, and the second polarizer 154 is located on the light detector side, that is, the objective lens 152 and the light detector. Located between 160. At this time, any one of the first polarizer 142 and the second polarizer 154 is a rotation polarizer is applied to rotate by using a motor or the like to adjust the polarization state. By adjusting one of the two polarizers constituting the polarizers 142 and 154 and analyzing the output image accordingly, the birefringence characteristics (phase delay of the biological cells, the main axis direction) of the living cells and tissues are measured. The polarizers 142 and 154 will be described later.

The phase delay unit 144 is provided between the first polarizer 142 and the second polarizer 154, and more specifically, is provided between the first polarizer 142 and the condenser lens 146. As the phase delay unit 144, a λ / 4 wave plate or the like may be used. The λ / 4 wave plate receives polarized light from the first polarizer 142 so that the phase difference between the slow moving axis (S.A .; Slow Axis) and the fast moving axis (F.A .; Fast Axis) is λ / 4.

The condenser lens 146 collects the light passing through the first polarizer 142 and the phase delay unit 144 to be concentrated on the sample of the measurement unit 150.

The measuring unit 150 allows the light passing through the first polarizer 142 and the phase delay unit 144 to pass through the sample for which the birefringence characteristic is to be measured. Although not shown, the measurement unit 150 may include a slide glass and a cover glass to allow a sample to be positioned therebetween.

The objective lens 152 enlarges the image of the light transmitted through the sample of the measuring unit 150 so as to be captured by the light detecting unit 160.

The second polarizer 154 is positioned between the objective lens 152 and the light detector 160, and is used to adjust the polarization state together with the first polarizer 142. In addition, the second polarizer 154 prevents a background image, which is light that does not pass through the sample, from passing through the measurement unit 150.

The light detector 160 converts the light passing through the second polarizer 154 into an electrical signal. A charge coupled device (CCD) camera or the like is used as the light detector 160 to detect the polarization state of the light passing through the sample as the light intensity.

FIG. 2 is a detailed conceptual diagram when the second polarizer adjusts polarization by rotating, and FIG. 3 is a detailed conceptual diagram when the first polarizer adjusts polarization by rotating. 2 and 3 are schematic diagrams of respective polarization microscope basic configurations according to which of the two polarizers the polarizer of the polarizer is adjusted.

Referring to FIG. 2, light generated from a light source passes through a first polarizer (fixed polarizer) and a λ / 4 wave plate, and then enters a CCD camera after polarization adjustment is performed through a second polarizer (rotating polarizer) through a sample. It is detected by the light intensity.

In the embodiment of FIG. 2, the polarization state of the output light incident on the CCD camera is expressed as Jones matrix.

In this case, A is the amplitude of incident light, ω is the frequency of incident light, and δ is the phase delay value by the sample.

In addition, the light intensity of the output light incident on the CCD camera is expressed by the following equation.

, δ는 샘플의 복굴절 특성 중 위상지연값, φ는 샘플의 복굴절 특성 중 주축의 방향, θ는 편광의 조정값, I_min은 출력광의 최소 광 강도, 즉 암 전류(dark current)에 의한 광 강도를 의미한다.here,

, δ is the phase retardation value of the birefringence characteristics of the sample, φ is the direction of the main axis of the birefringence characteristics of the sample, θ is the adjustment value of the polarization, I _min is the minimum light intensity of the output light, that is, the light intensity due to dark current Means.

Referring to FIG. 3, light generated from a light source is polarized light adjusted through a first polarizer (rotating polarizer), passes through a λ / 4 wave plate and a second polarizer (fixed polarizer) through a sample, and then enters a CCD camera. And light intensity is detected.

In the embodiment of FIG. 3, the polarization state of the output light incident to the CCD camera is expressed as Jones matrix.

In the embodiment of FIG. 3, the light intensity of the output light incident on the CCD camera is the same as that in Equation 2 above.

4 is a graph of light intensity finally obtained by the light detector through polarization adjustment.

Of the two polarizers constituting the polarizer, the polarization state of one polarizer may be adjusted and the light intensity according to the polarizer may be collected to obtain a sine waveform light intensity graph as illustrated in FIG. 4. In FIG. 4, the horizontal axis represents polarization adjustment value θ, and the vertical axis represents light intensity I of output light. In FIG. 4, θ _peak = θ _IMax , that is, the polarization adjustment value when the light intensity reaches the maximum, I _Max means the maximum light intensity of the output light, and I _Min means the minimum light intensity of the output light.

The birefringence characteristics of the sample can be measured by substituting the I _Max , I _Min , and θ _IMax values obtained through FIG. 4 into the following equation.

Each of the above Equations 4 and 5 is obtained by obtaining the main axis direction φ and the phase delay value δ of the birefringence characteristics of the sample. As described above, the birefringence characteristic of the sample can be measured by using the sine function-type light intensity obtained by the light detection unit and the equations (4) and (5) through polarization adjustment.

5 and 6 are detailed conceptual diagrams when polarization is adjusted using a liquid crystal variable phase delayer instead of rotating a linear polarizer using a motor as shown in FIGS. 2 and 3.

In addition to rotating the linear polarizer using a motor, a liquid crystal variable phase retarder may be provided between the fixed linear polarizer and the λ / 4 wave plate. There is a method of controlling the liquid crystal variable phase delay unit by an electric signal.

The liquid crystal variable phase delay unit may have a configuration in which a liquid crystal is inserted between a pair of flat glass plates coated with transparent electrodes on opposite inner surfaces thereof. When no voltage is applied through the transparent electrode, the liquid crystals are arranged in a line along the glass plate, and the phase delay of incident light is maximized. As the voltage applied through the transparent electrode increases, the liquid crystal forms an oblique angle with the glass plate, and the angle between the electric field direction and the longitudinal direction of the liquid crystal becomes smaller. As a result, the phase delay of incident light becomes smaller. When a voltage above a certain threshold is applied through the transparent electrode, the electric field direction and the longitudinal direction of the liquid crystal are parallel to each other, and the phase delay of incident light is minimized. The liquid crystal variable phase delay unit can simply control the phase delay of the polarized light passing through the polarizer by controlling the electrical signal or the voltage.

Referring to FIG. 5, light generated from a light source passes through a polarizer (fixed polarizer) and then reaches a λ / 4 wave plate through a liquid crystal variable phase delay unit. Although not shown, the light passing through the [lambda] / 4 wave plate is incident on the CCD camera through the sample and detected by the light intensity. In FIG. 5, for example, the transmission axis TA of the polarizer coincides with the slow propagating axis SA of the λ / 4 wave plate, and the slow propagating axis SA of the liquid crystal variable phase delay unit is defined as the transmission axis of the polarizer. The arrangement is made by rotating -π / 4 relative to TA).

In the embodiment of FIG. 5, the polarization state of the output light incident on the CCD camera is expressed as Jones matrix.

6, the transmission axis TA of the polarizer coincides with the fast-moving axis FA of the λ / 4 wave plate, and the fast-moving axis FA of the liquid crystal variable phase delay unit is aligned with the transmission axis TA of the polarizer. An example in which the array is rotated with respect to -π / 4 is shown.

In the embodiment of FIG. 6, the polarization state of the output light incident on the CCD camera is expressed as Jones matrix.

In Equations 6 and 7, θ represents a phase delay value of the liquid crystal variable phase delay unit.

7 is a graph illustrating an electrical signal supplied to a liquid crystal variable phase delay unit and a phase delay value according to an amplitude of the electrical signal. 7A illustrates a voltage supplied to the liquid crystal variable phase delay unit as time passes, and FIG. 7B illustrates a phase delay value according to the voltage supplied to the liquid crystal variable phase delay unit.

The liquid crystal variable phase delay unit may be supplied with an electric signal of several MHz. Referring to FIG. 7, it can be seen that the phase delay value can be controlled by adjusting the amplitude (eg, voltage) of the electrical signal supplied to the liquid crystal variable phase delay unit. By adjusting the phase delay value of the liquid crystal variable phase retarder as described above, the same effect as adjusting the polarization by rotating the linear polarizer using a motor can be obtained (see Equations 6 and 7). Furthermore, since the liquid crystal variable phase delay can be controlled through an electrical signal having a frequency of several MHz, the polarization can be controlled at a higher speed than when the polarization is adjusted by rotation by a motor, thereby realizing birefringence characteristics in real time. Since it can be measured, it is possible to characterize the metastasis and differentiation of cells by changing the birefringence properties during metastasis and differentiation of cells.

As such, by adjusting the polarization state of the polarization microscope, the light intensity accordingly can be collected to measure the birefringence characteristics of the sample including the direction and the phase delay value of the main axis. By measuring the birefringence characteristics, the birefringence characteristics of normal cells and cancer cells can be identified to diagnose cancer cells, and the metastasis and differentiation characteristics of the cells can be identified.

The above description is merely illustrative of the technical idea of the present invention, and various modifications, changes and substitutions may be made by those skilled in the art without departing from the essential characteristics of the present invention. It will be possible. Accordingly, the embodiments disclosed in the present invention and the accompanying drawings are not intended to limit the technical spirit of the present invention but to describe the present invention, and the scope of the technical idea of the present invention is not limited by the embodiments and the accompanying drawings. . The protection scope of the present invention should be interpreted by the following claims, and all technical ideas within the equivalent scope should be interpreted as being included in the scope of the present invention.

The present invention can be widely used in the life / medical field such as diagnosis of cancer cells and cancer tissues, observation of metastasis and differentiation characteristics of cells.

1 is a block diagram of a medical polarization microscope system according to an embodiment of the present invention,

2 is a detailed conceptual diagram in the case where the second polarizer adjusts polarization by rotating;

3 is a detailed conceptual diagram in the case where the first polarizer adjusts the polarization by rotating;

4 is a graph of the light intensity finally obtained by the light detection unit through the polarization adjustment,

5 and 6 are detailed conceptual diagrams in the case of adjusting polarization using a liquid crystal variable phase delay unit;

7 is a graph illustrating an electrical signal supplied to a liquid crystal variable phase delay unit and a phase delay value according to an amplitude of the electrical signal.

<Explanation of symbols for the main parts of the drawings>

100-Medical Polarization Microscope System 110-Light Generator

120-Lens part 142, 154-Polarizing part

144-Phase delay unit 150-Measurement unit

160-light detector

Claims (14)

A light generator for generating light; A lens unit converting the generated light into parallel light; A polarizer which adjusts a polarization state of light incident from the lens unit and includes a first polarizer and a second polarizer; A phase delay unit provided between the first polarizer and the second polarizer; A measuring unit configured to pass the light passing through the first polarizer through a sample to be measured; And A light detector for detecting the polarization state of the light passing through the sample as the light intensity Medical polarizing microscope system by polarization adjustment comprising a. The method of claim 1, The medical polarizing microscope system according to polarization adjustment, wherein any one of the first polarizer and the second polarizer is a rotating polarizer. The method of claim 1, The phase retardation unit comprises a λ / 4 wave plate, characterized in that the medical polarization microscope system by polarization adjustment. The method of claim 1, The optical detection unit comprises a CCD (Charge Coupled Device) camera for converting the light passing through the sample into an electrical signal, characterized in that the medical polarization microscope system by polarization adjustment. The method of claim 1, The lens unit includes an optical diffuser for suppressing spatial coherence of the light emitted from the light generator, and a plano-convex lense for converting the light passing through the diffuser into parallel light. Medical polarized light microscope system by polarization adjustment characterized by the above-mentioned. A light generator for generating light; A lens unit converting the generated light into parallel light; A polarizer including a polarizer generating polarized light by receiving parallel light from the lens unit, and a liquid crystal variable phase retarder for adjusting the polarized light by controlling a phase delay of the polarized light by an electric signal; A measuring unit which passes the light passing through the polarizing unit to the sample to be measured; And A light detector for detecting the polarization state of the light passing through the sample as the light intensity Medical polarization microscope system by polarization adjustment comprising a. The method of claim 6, And a λ / 4 wave plate is provided between the liquid crystal variable phase delay unit and the measurement unit. The method of claim 7, wherein The transmission axis TA of the polarizer coincides with the slow propagating axis SA of the λ / 4 wave plate, and the slow propagating axis SA of the liquid crystal variable phase delay unit is the transmission axis TA of the polarizer. Medical polarization microscope system by polarization adjustment characterized in that it arranged by rotating -π / 4 relative to. The method of claim 7, wherein The transmission axis TA of the polarizer coincides with the rapidly advancing axis FA of the λ / 4 wave plate, and the rapid advancing axis FA of the liquid crystal variable phase delay unit is the transmission axis TA of the polarizer. Medical polarization microscope system by polarization adjustment characterized in that it arranged by rotating -π / 4 relative to. The method of claim 6, The optical detection unit comprises a CCD (Charge Coupled Device) camera for converting the light passing through the sample into an electrical signal, characterized in that the medical polarization microscope system by polarization adjustment. The method of claim 6, The lens unit includes an optical diffuser for suppressing spatial coherence of the light emitted from the light generator, and a plano-convex lense for converting the light passing through the diffuser into parallel light. Medical polarized light microscope system by polarization adjustment characterized by the above-mentioned. (a) generating light; (b) converting the generated light into parallel light; (c) adjusting the polarization state of the sample whose birefringence characteristic is to be measured; And (d) measuring the birefringence characteristics of the sample including the direction and the phase delay value of the main axis by collecting the light intensity according to the polarization adjustment Method for measuring the birefringence characteristics by polarization adjustment comprising a. The method of claim 12, The direction (phi) of the main axis of the step (d) is by the following equation, characterized in that for measuring the birefringence characteristics by polarization adjustment.

here,

Is the polarization adjustment value when the light intensity of the output light is maximum. The method of claim 12, The phase delay value δ of step (d) is determined by the following equation, wherein the birefringence characteristic is measured by polarization adjustment.

Where I _max is the maximum light intensity of the output light and I _min is the minimum light intensity of the output light

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