JP5098067B2 - Quantification standardized microscope equipment - Google Patents

Description

The present invention is a technique related to quantification in microscopic measurement.

In the conventional quantification technique, a grid is marked on a slide glass or a cover glass like a hemocytometer, and the quantification is performed based on the range and depth of the grid.

Conventional quantification techniques required the marking of a grid on a slide glass or cover glass. Therefore, in order to change the lattice spacing, it is necessary to prepare a slide glass or cover glass on which another lattice is marked, and various lattices cannot be used with the same sample. In addition, since the grid is marked on the slide glass or the cover glass, it takes a lot of time for manufacturing, and therefore, it is very expensive. Furthermore, there was no quantification technique other than the lattice.

1 [Means of Claim 1]
The quantified standardized microscope apparatus according to claim 1 is:
An XY stage means for holding a sample collected from a living body and moving it at least in the XY direction in order to observe each part of the sample;

Sample imaging means for imaging sample image information from the sample, and a sample (pixel) block image means for obtaining a two-dimensional image block composed of pixels consisting of resolution units from the sample imaging means and the XY stage means Index image capturing means for capturing the imaged index, and index (pixel) block image means for obtaining a two-dimensional image block composed of pixels comprising resolution units from the index image capturing means;


A correction grid having a constant pitch installed on the XY stage means;
In the index (pixel) block image obtained by imaging the correction grid,
By taking a slightly different number of correction grids as a video image with respect to the number of blocks in the block image, the image pickup pixel pitch and the index (pixel) block image pitch are slightly shifted, resulting in the beat obtained by the shift amount. High-resolution relative position information obtained by capturing image amplitude
Or
(Absolute) position information from the index image obtained in the same manner as described above using a pitch change correction grid having a spatial frequency change as the correction grid
Use one or a combination of
At least to improve positioning resolution or / and
At least by the electron lattice correction means to improve the resolution of the electron lattice,

The both (pixel) block images are
Based on the amplitude interval of the beat image in the x direction of the X stage means obtained by the electronic lattice correction means from the correction grid imaged on the index pixel block, the electronic grid means performs a grid interval on the sample pixel block. The interval setting
A mechanism based on relative positioning between the two pixel blocks obtained by setting in the same way for the y direction in the Y stage ;

A mechanism constituted by absolute positioning of the two pixel blocks obtained from (absolute) position information from the index image obtained in the same manner using the pitch change correction grid.
In a mechanism comprising either or a combination of


At least one point index installed on the XY stage means;

The coordinates of the point index are (index) a coordinate detection means that uses either the center of gravity of the final convergence block of the light spot tracking means (device) or individual pixels.

It is characterized by providing.

2 [Means of claim 2]
The quantified standardized microscope apparatus according to claim 2 is:
An XY stage means for holding a sample collected from a living body and moving it at least in the XY direction in order to observe each part of the sample;

Sample imaging means for imaging sample image information from the sample, and a sample (pixel) block image means for obtaining a two-dimensional image block composed of pixels consisting of resolution units from the sample imaging means and the XY stage means Index image capturing means for capturing the imaged index, and index (pixel) block image means for obtaining a two-dimensional image block composed of pixels comprising resolution units from the index image capturing means;


A correction grid having a constant pitch installed on the XY stage means;
In the index (pixel) block image obtained by imaging the correction grid,
By taking a slightly different number of correction grids as a video image with respect to the number of blocks in the block image, the image pickup pixel pitch and the index (pixel) block image pitch are slightly shifted, resulting in the beat obtained by the shift amount. High-resolution relative position information obtained by capturing image amplitude
Or
(Absolute) position information from the index image obtained in the same manner as described above using a pitch change correction grid having a spatial frequency change as the correction grid
Use one or a combination of
At least to improve positioning resolution or / and
At least by the electron lattice correction means to improve the resolution of the electron lattice,

The both (pixel) block images are
Based on the amplitude interval of the beat image in the x direction of the X stage means obtained by the electronic lattice correction means from the correction grid imaged on the index pixel block, the electronic grid means performs a grid interval on the sample pixel block. The interval setting
A mechanism based on relative positioning between the two pixel blocks obtained by setting in the same way for the y direction in the Y stage ;

A mechanism constituted by absolute positioning of the two pixel blocks obtained from (absolute) position information from the index image obtained in the same manner using the pitch change correction grid.
In a mechanism comprising either or a combination of


At least one point index installed on the XY stage means;

A coordinate detection means that uses the coordinates of the point index as the (index) center of gravity of the final convergence block of the light spot tracking means (device) or individual pixels;

An electron grid means for generating an electron grid based on the index position information from the coordinate detection means means;
It is characterized by providing.

3 (Means of claim 3)
The quantified standardized microscope apparatus according to claim 3 is:
An XY stage means for holding a sample collected from a living body and moving it at least in the XY direction in order to observe each part of the sample;

Sample imaging means for imaging sample image information from the sample, and a sample (pixel) block image means for obtaining a two-dimensional image block composed of pixels consisting of resolution units from the sample imaging means and the XY stage means Index image capturing means for capturing the imaged index, and index (pixel) block image means for obtaining a two-dimensional image block composed of pixels comprising resolution units from the index image capturing means;


A correction grid having a constant pitch installed on the XY stage means;
In the index (pixel) block image obtained by imaging the correction grid,
By taking a slightly different number of correction grids as a video image with respect to the number of blocks in the block image, the image pickup pixel pitch and the index (pixel) block image pitch are slightly shifted, resulting in the beat obtained by the shift amount. High-resolution relative position information obtained by capturing image amplitude
Or
(Absolute) position information from the index image obtained in the same manner as described above using a pitch change correction grid having a spatial frequency change as the correction grid
Use one or a combination of
At least to improve positioning resolution or / and
At least by the electron lattice correction means to improve the resolution of the electron lattice,

The both (pixel) block images are
Based on the amplitude interval of the beat image in the x direction of the X stage means obtained by the electronic lattice correction means from the correction grid imaged on the index pixel block, the electronic grid means performs a grid interval on the sample pixel block. The interval setting
A mechanism based on relative positioning between the two pixel blocks obtained by setting in the same way for the y direction in the Y stage ;

A mechanism constituted by absolute positioning of the two pixel blocks obtained from (absolute) position information from the index image obtained in the same manner using the pitch change correction grid.
In a mechanism comprising either or a combination of


At least one point index installed on the XY stage means;

A coordinate detection means that uses the coordinates of the point index as the (index) center of gravity of the final convergence block of the light spot tracking means (device) or individual pixels;

An electron grid means for generating an electron grid based on index position information from the coordinate detection means means;

Overlaying means (overlap means) for superimposing the electron lattice and the sample image,
It is characterized by providing.

4 (Means of claim 4)
The quantified standardized microscope apparatus according to claim 4 is:
An XY stage means for holding a sample collected from a living body and moving it at least in the XY direction in order to observe each part of the sample;

Sample imaging means for imaging sample image information from the sample, and a sample (pixel) block image means for obtaining a two-dimensional image block composed of pixels consisting of resolution units from the sample imaging means and the XY stage means Index image capturing means for capturing the imaged index, and index (pixel) block image means for obtaining a two-dimensional image block composed of pixels comprising resolution units from the index image capturing means;


A correction grid having a constant pitch installed on the XY stage means;
In the index (pixel) block image obtained by imaging the correction grid,
By taking a slightly different number of correction grids as a video image with respect to the number of blocks in the block image, the image pickup pixel pitch and the index (pixel) block image pitch are slightly shifted, resulting in the beat obtained by the shift amount. High-resolution relative position information obtained by capturing image amplitude
Or
(Absolute) position information from the index image obtained in the same manner as described above using a pitch change correction grid having a spatial frequency change as the correction grid
Use one or a combination of
At least to improve positioning resolution or / and
At least by the electron lattice correction means to improve the resolution of the electron lattice,

The both (pixel) block images are
Based on the amplitude interval of the beat image in the x direction of the X stage means obtained by the electronic lattice correction means from the correction grid imaged on the index pixel block, the electronic grid means performs a grid interval on the sample pixel block. The interval setting
A mechanism based on relative positioning between the two pixel blocks obtained by setting in the same way for the y direction in the Y stage ;

A mechanism constituted by absolute positioning of the two pixel blocks obtained from (absolute) position information from the index image obtained in the same manner using the pitch change correction grid.
In a mechanism comprising either or a combination of


A correction grating having a constant pitch installed in the XY stage means;
In the index (pixel) block image obtained by imaging the correction grid,
By taking a slightly different number of correction grids as a video image with respect to the number of blocks in the block image, the image pickup pixel pitch and the index (pixel) block image pitch are slightly shifted, resulting in the beat obtained by the shift amount. High-resolution relative position information obtained by capturing the amplitude of the image, or
Using any one or a combination of (absolute) position information from the index image obtained in the same manner as described above using a pitch change correction grid having a spatial frequency change as the correction grid ,
At least to improve positioning resolution or / and
An electron lattice correction means for improving the resolution of at least the electron lattice,
It is characterized by providing.

5 (Means of Claim 5)
The quantified standardized microscope apparatus according to claim 5 is:
An XY stage means for holding a sample collected from a living body and moving it at least in the XY direction in order to observe each part of the sample;

Sample imaging means for imaging sample image information from the sample, and a sample (pixel) block image means for obtaining a two-dimensional image block composed of pixels consisting of resolution units from the sample imaging means and the XY stage means Index image capturing means for capturing the imaged index, and index (pixel) block image means for obtaining a two-dimensional image block composed of pixels comprising resolution units from the index image capturing means;


A correction grid having a constant pitch installed on the XY stage means;
In the index (pixel) block image obtained by imaging the correction grid,
By taking a slightly different number of correction grids as a video image with respect to the number of blocks in the block image, the image pickup pixel pitch and the index (pixel) block image pitch are slightly shifted, resulting in the beat obtained by the shift amount. High-resolution relative position information obtained by capturing image amplitude
Or
(Absolute) position information from the index image obtained in the same manner as described above using a pitch change correction grid having a spatial frequency change as the correction grid
Use one or a combination of
At least to improve positioning resolution or / and
At least by the electron lattice correction means to improve the resolution of the electron lattice,

The both (pixel) block images are
Based on the amplitude interval of the beat image in the x direction of the X stage means obtained by the electronic lattice correction means from the correction grid imaged on the index pixel block, the electronic grid means performs a grid interval on the sample pixel block. The interval setting
A mechanism based on relative positioning between the two pixel blocks obtained by setting in the same way for the y direction in the Y stage ;

A mechanism constituted by absolute positioning of the two pixel blocks obtained from (absolute) position information from the index image obtained in the same manner using the pitch change correction grid.
In a mechanism comprising either or a combination of


A correction grid having a constant pitch installed in the XY stage means and at least one point index installed in the XY stage means;

Coordinate detection means for detecting index coordinates based on the point index;

Absolute position information obtained by the index;
Or
In the index (pixel) block image obtained by imaging the correction grid, the number of correction grids slightly different from the number of blocks in the block image is captured as a video, whereby the imaging pixel pitch and the index (pixel) block image are captured. High-resolution relative position information obtained by capturing the amplitude of the beat image obtained by the amount of the deviation due to the slight deviation of the pitch,
Or
(Absolute) position information from the index image obtained in the same manner as described above using a pitch change correction grid having a spatial frequency change in the correction grid;
Use one or a combination of
At least to improve positioning resolution or / and
An electron lattice correction means for improving the resolution of at least the electron lattice,
It is characterized by providing.

6 (Means of claim 6)
The quantified standardized microscope apparatus according to claim 6 is:
A continuous probe is provided as a collection and quantification means for quantifying a sample collected from a living body or the like.

7 (Means of Claim 7)
The quantified standardized microscope apparatus according to claim 7 is:
A quantification standardized microscope apparatus comprising an image recognition means for recognizing a specific bacterium.

1 [Operation and effect of claim 1]
The quantified standardized microscope apparatus according to claim 1 is:
An XY stage means for holding a sample collected from a living body and moving it at least in the XY direction in order to observe each part of the sample;

Sample imaging means for imaging sample image information from the sample, and a sample (pixel) block image means for obtaining a two-dimensional image block composed of pixels consisting of resolution units from the sample imaging means and the XY stage means Index image capturing means for capturing the imaged index, and index (pixel) block image means for obtaining a two-dimensional image block composed of pixels comprising resolution units from the index image capturing means;


A correction grid having a constant pitch installed on the XY stage means;
In the index (pixel) block image obtained by imaging the correction grid,
By taking a slightly different number of correction grids as a video image with respect to the number of blocks in the block image, the image pickup pixel pitch and the index (pixel) block image pitch are slightly shifted, resulting in the beat obtained by the shift amount. High-resolution relative position information obtained by capturing image amplitude
Or
(Absolute) position information from the index image obtained in the same manner as described above using a pitch change correction grid having a spatial frequency change as the correction grid
Use one or a combination of
At least to improve positioning resolution or / and
At least by the electron lattice correction means to improve the resolution of the electron lattice,

The both (pixel) block images are
Based on the amplitude interval of the beat image in the x direction of the X stage means obtained by the electronic lattice correction means from the correction grid imaged on the index pixel block, the electronic grid means performs a grid interval on the sample pixel block. The interval setting
A mechanism based on relative positioning between the two pixel blocks obtained by setting in the same way for the y direction in the Y stage ;

A mechanism constituted by absolute positioning of the two pixel blocks obtained from (absolute) position information from the index image obtained in the same manner using the pitch change correction grid.
In a mechanism comprising either or a combination of


At least one point index installed on the XY stage means;

The center of gravity of the last converging blocks of the coordinates of the point indicators (indicators) point tracking means (device), so it characterized that and a coordinate detection unit to use either of the individual pixels,
A resolution smaller than the resolution R of the conventional positioning accuracy can be obtained. This improves positioning accuracy and various sample measurements.

2 [Operation and effect of claim 2]
The quantified standardized microscope apparatus according to claim 2 is:
An XY stage means for holding a sample collected from a living body and moving it at least in the XY direction in order to observe each part of the sample;

Sample imaging means for imaging sample image information from the sample, and a sample (pixel) block image means for obtaining a two-dimensional image block composed of pixels consisting of resolution units from the sample imaging means and the XY stage means Index image capturing means for capturing the imaged index, and index (pixel) block image means for obtaining a two-dimensional image block composed of pixels comprising resolution units from the index image capturing means;


A correction grid having a constant pitch installed on the XY stage means;
In the index (pixel) block image obtained by imaging the correction grid,
By taking a slightly different number of correction grids as a video image with respect to the number of blocks in the block image, the image pickup pixel pitch and the index (pixel) block image pitch are slightly shifted, resulting in the beat obtained by the shift amount. High-resolution relative position information obtained by capturing image amplitude
Or
(Absolute) position information from the index image obtained in the same manner as described above using a pitch change correction grid having a spatial frequency change as the correction grid
Use one or a combination of
At least to improve positioning resolution or / and
At least by the electron lattice correction means to improve the resolution of the electron lattice,

The both (pixel) block images are
Based on the amplitude interval of the beat image in the x direction of the X stage means obtained by the electronic lattice correction means from the correction grid imaged on the index pixel block, the electronic grid means performs a grid interval on the sample pixel block. The interval setting
A mechanism based on relative positioning between the two pixel blocks obtained by setting in the same way for the y direction in the Y stage ;

A mechanism constituted by absolute positioning of the two pixel blocks obtained from (absolute) position information from the index image obtained in the same manner using the pitch change correction grid.
In a mechanism comprising either or a combination of


At least one point index installed on the XY stage means;

A coordinate detection means that uses the coordinates of the point index as the (index) center of gravity of the final convergence block of the light spot tracking means (device) or individual pixels;

An electron grid means for generating an electron grid based on the index position information from the coordinate detection means means;
Since it is characterized by providing,
A resolution smaller than the resolution R of the conventional positioning accuracy can be obtained. This improves the positioning accuracy. Further, the position, shape, depth, and the like of the sample can be measured with higher accuracy by using the electronic lattice obtained from the accurate position information.

3 [Operation and effect of claim 3]
The quantified standardized microscope apparatus according to claim 3 is:
An XY stage means for holding a sample collected from a living body and moving it at least in the XY direction in order to observe each part of the sample;

Sample imaging means for imaging sample image information from the sample, and a sample (pixel) block image means for obtaining a two-dimensional image block composed of pixels consisting of resolution units from the sample imaging means and the XY stage means Index image capturing means for capturing the imaged index, and index (pixel) block image means for obtaining a two-dimensional image block composed of pixels comprising resolution units from the index image capturing means;


A correction grid having a constant pitch installed on the XY stage means;
In the index (pixel) block image obtained by imaging the correction grid,
By taking a slightly different number of correction grids as a video image with respect to the number of blocks in the block image, the image pickup pixel pitch and the index (pixel) block image pitch are slightly shifted, resulting in the beat obtained by the shift amount. High-resolution relative position information obtained by capturing image amplitude
Or
(Absolute) position information from the index image obtained in the same manner as described above using a pitch change correction grid having a spatial frequency change as the correction grid
Use one or a combination of
At least to improve positioning resolution or / and
At least by the electron lattice correction means to improve the resolution of the electron lattice,

The both (pixel) block images are
Based on the amplitude interval of the beat image in the x direction of the X stage means obtained by the electronic lattice correction means from the correction grid imaged on the index pixel block, the electronic grid means performs a grid interval on the sample pixel block. The interval setting
A mechanism based on relative positioning between the two pixel blocks obtained by setting in the same way for the y direction in the Y stage ;

A mechanism constituted by absolute positioning of the two pixel blocks obtained from (absolute) position information from the index image obtained in the same manner using the pitch change correction grid.
In a mechanism comprising either or a combination of


At least one point index installed on the XY stage means;

A coordinate detection means that uses the coordinates of the point index as the (index) center of gravity of the final convergence block of the light spot tracking means (device) or individual pixels;

An electron grid means for generating an electron grid based on index position information from the coordinate detection means means;

Overlaying means (overlap means) for superimposing the electron lattice and the sample image,
Since it is characterized by providing,
A resolution smaller than the resolution R of the conventional positioning accuracy can be obtained. This improves the positioning accuracy. In addition, measurement by comparison with the sample image can be performed with higher accuracy by the electron grid obtained from the position information with high accuracy.

4 [Operation and effect of claim 4]
The quantified standardized microscope apparatus according to claim 4 is:
An XY stage means for holding a sample collected from a living body and moving it at least in the XY direction in order to observe each part of the sample;

Sample imaging means for imaging sample image information from the sample, and a sample (pixel) block image means for obtaining a two-dimensional image block composed of pixels consisting of resolution units from the sample imaging means and the XY stage means Index image capturing means for capturing the imaged index, and index (pixel) block image means for obtaining a two-dimensional image block composed of pixels comprising resolution units from the index image capturing means;


A correction grid having a constant pitch installed on the XY stage means;
In the index (pixel) block image obtained by imaging the correction grid,
By taking a slightly different number of correction grids as a video image with respect to the number of blocks in the block image, the image pickup pixel pitch and the index (pixel) block image pitch are slightly shifted, resulting in the beat obtained by the shift amount. High-resolution relative position information obtained by capturing image amplitude
Or
(Absolute) position information from the index image obtained in the same manner as described above using a pitch change correction grid having a spatial frequency change as the correction grid
Use one or a combination of
At least to improve positioning resolution or / and
At least by the electron lattice correction means to improve the resolution of the electron lattice,

The both (pixel) block images are
Based on the amplitude interval of the beat image in the x direction of the X stage means obtained by the electronic lattice correction means from the correction grid imaged on the index pixel block, the electronic grid means performs a grid interval on the sample pixel block. The interval setting
A mechanism based on relative positioning between the two pixel blocks obtained by setting in the same way for the y direction in the Y stage ;

A mechanism constituted by absolute positioning of the two pixel blocks obtained from (absolute) position information from the index image obtained in the same manner using the pitch change correction grid.
In a mechanism comprising either or a combination of


A correction grating having a constant pitch installed in the XY stage means;
In the index (pixel) block image obtained by imaging the correction grid,
By taking a slightly different number of correction grids as a video image with respect to the number of blocks in the block image, the image pickup pixel pitch and the index (pixel) block image pitch are slightly shifted, resulting in the beat obtained by the shift amount. High-resolution relative position information obtained by capturing the amplitude of the image, or
Using any one or a combination of (absolute) position information from the index image obtained in the same manner as described above using a pitch change correction grid having a spatial frequency change as the correction grid ,
At least to improve positioning resolution or / and
An electron lattice correction means for improving the resolution of at least the electron lattice,
Since it is characterized by providing,
The electronic lattice correction means can obtain a resolution of about one digit smaller than the conventional resolution R of the positioning accuracy. This improves positioning accuracy and various sample measurements.


5 [Operation and effect of claim 5]
The quantified standardized microscope apparatus according to claim 5 is:
An XY stage means for holding a sample collected from a living body and moving it at least in the XY direction in order to observe each part of the sample;

Sample imaging means for imaging sample image information from the sample, and a sample (pixel) block image means for obtaining a two-dimensional image block composed of pixels consisting of resolution units from the sample imaging means and the XY stage means Index image capturing means for capturing the imaged index, and index (pixel) block image means for obtaining a two-dimensional image block composed of pixels comprising resolution units from the index image capturing means;


A correction grid having a constant pitch installed on the XY stage means;
In the index (pixel) block image obtained by imaging the correction grid,
By taking a slightly different number of correction grids as a video image with respect to the number of blocks in the block image, the image pickup pixel pitch and the index (pixel) block image pitch are slightly shifted, resulting in the beat obtained by the shift amount. High-resolution relative position information obtained by capturing image amplitude
Or
(Absolute) position information from the index image obtained in the same manner as described above using a pitch change correction grid having a spatial frequency change as the correction grid
Use one or a combination of
At least to improve positioning resolution or / and
At least by the electron lattice correction means to improve the resolution of the electron lattice,

The both (pixel) block images are
Based on the amplitude interval of the beat image in the x direction of the X stage means obtained by the electronic lattice correction means from the correction grid imaged on the index pixel block, the electronic grid means performs a grid interval on the sample pixel block. The interval setting
A mechanism based on relative positioning between the two pixel blocks obtained by setting in the same way for the y direction in the Y stage ;

A mechanism constituted by absolute positioning of the two pixel blocks obtained from (absolute) position information from the index image obtained in the same manner using the pitch change correction grid.
In a mechanism comprising either or a combination of


A correction grid having a constant pitch installed in the XY stage means and at least one point index installed in the XY stage means;

Coordinate detection means for detecting index coordinates based on the point index;

Absolute position information obtained by the index;
Or
In the index (pixel) block image obtained by imaging the correction grid, the number of correction grids slightly different from the number of blocks in the block image is captured as a video, whereby the imaging pixel pitch and the index (pixel) block image are captured. High-resolution relative position information obtained by capturing the amplitude of the beat image obtained by the amount of the deviation due to the slight deviation of the pitch,
Or
(Absolute) position information from the index image obtained in the same manner as described above using a pitch change correction grid having a spatial frequency change in the correction grid;
Use one or a combination of
At least to improve positioning resolution or / and
An electron lattice correction means for improving the resolution of at least the electron lattice,
Since it is characterized by providing,
By using the electron lattice correction means and the light spot tracking index detection, it is possible to obtain a resolution that is smaller by one digit or more than the conventional resolution R of positioning accuracy, smoothness and ease of positioning. This improves positioning accuracy, various sample measurements, operability, and the like.

6 [Operation and effect of claim 6]
Quantification standardized microscope equipment
Since, characterized in that it comprises a continuous circular probe as a collecting quantitative means to quantify the collected samples from such organism, by quantitative and continuously sampled samples by continuous circle, volume, cell number, the number Speed can be measured quantitatively.

7 [Operation and effect of claim 7]
The quantified standardized microscope apparatus according to claim 7 is:
Since it is equipped with image recognition means for recognizing specific bacteria ,
P. I. Specific bacteria such as and Stm can be measured, tested, and diagnosed.



Action and Effect 1 of the Old Invention [Action and Effect of Claim 1]
The quantification standardization (phase difference) microscope apparatus according to claim 1 is:
Since it is characterized by comprising index means and index detection means,
Quantitative measurement is possible. By this quantitative measurement, various information such as the length, area, number, size, speed, acceleration, and movement of the sample can be obtained.
Further, the position of the sample stage can be detected by the index detection means.
Thereby, an image corresponding to the sample position can be detected and searched at any time.

2 [Operation and effect of claim 2]
The quantification standardization (phase difference) microscope apparatus according to claim 2 is:
By using the index means, the index detection means, etc.
Since it is characterized by comprising quantitative measurement means for performing (quantitative) measurement on an observation object such as microorganisms, tissues, red blood cells, and white blood cells, (quantitative) measurement can be performed.
By this quantitative measurement, various information such as the length, area, number, size, speed, acceleration, and movement of the sample can be obtained. In particular, one-dimensional or two-dimensional measurement can be measured with high accuracy.

3 [Operation and effect of claim 3]
Since the quantification standardization (phase difference) microscope apparatus according to the third aspect includes a three-dimensional quantitative measurement means provided with either a cover glass or a slide glass having a known depth, three-dimensional quantitative measurement such as density can be performed.

4 [Operation and effect of claim 4]
The quantification standardization (phase difference) microscope apparatus according to claim 4 is characterized by comprising a three-dimensional quantitative measurement means provided with a thickness measurement means capable of measuring the thickness of the sample, so that three-dimensional quantitative measurement such as density can be performed. .

5 [Operation and effect of claim 5]
Since the quantification standardization (phase difference) microscope apparatus according to the fifth aspect includes the electronic lattice correction means, more accurate quantitative measurement can be performed.
Furthermore, it is not necessary to use a cover glass with a lattice or a slide glass with a lattice. As a result, the running cost is low, and there is no phase disturbance or image quality degradation due to the grating.

6 [Operation and effect of claim 6]
The quantification standardization (phase difference) microscope apparatus according to claim 6 is:
Since the collection quantification means is provided, the accuracy is further improved because the collected material is quantified. In addition, the liquid phase (including flow and semi-flow) and plaque phase (layer) of the periodontal pocket can be selected and collected according to the collection shape of the collection and quantification means. Samples can also be collected from any position in the periodontal pocket.
Furthermore, quantitative sampling is possible.
As a result, the absolute value of the population can be measured, which further increases the accuracy.

7 [Operation and effect of claim 7]
The quantification standardization (phase difference) microscope apparatus according to claim 7 is:
Since it is characterized by having at least electromagnetic wave analysis by fluorescence means or spectroscopic means, or property means by motion analysis by motion analysis means, etc., it is suitable for the properties (spectral characteristics, fluorescence characteristics, motion characteristics, etc.) of observation objects such as microorganisms. Thus, an observation object such as a microorganism in the sample image can be specified.

Quantification normalized microscope apparatus of the present invention, will be described with reference to embodiments or variations shown in FIGS. 1-8.

[Configuration of Example]
Quantification normalized microscope apparatus in Fig. 1,
At least an electronic lattice means is provided.

[Operation]
When the power is turned on, an electronic grid is displayed on the sample image. 4 lower figure, FIG. 8, etc.

[effect]
The size of the sample can be determined by the electron lattice.

[Modification]
In addition to the electronic lattice, an electronic scale such as a ruler scale, a protractor scale, a circular scale, a triangular scale, an arc scale, and a polygon scale may be displayed.
In addition, an electronic lattice correction means described later may be provided to improve the accuracy and movement of the lattice.

Quantification normalized microscope apparatus of the first embodiment is provided with at least quantitatively measuring means.

[First Embodiment]
The first embodiment is directed to embodiments quantification measurement by quantification normalized microscope apparatus.

[Constitution]

The quantified standardized phase contrast microscope apparatus in FIG.

It is characterized by having a quantitative measurement means .

Here, the quantitative measurement means is at least

The sample stage means (preferably, provided with an index means serving as an index for obtaining electronic lattice information for a sample image, and further having a sample holding means for holding a sample as an object to be measured. A stage whose position can be changed in one direction of X, two directions in XY, or three directions of XYZ is desirable).



Index detecting means for detecting an index (means) provided in the sample stage (means) and detecting electronic lattice information for drawing (generating) an electronic lattice on a sample image based on the index ;



An electron lattice means for generating an electron lattice of a specific size and shape for the sample image according to the electron lattice information (mainly absolute position information or relative position information of the electron lattice with respect to the sample image);



Sample imaging means for imaging the sample held on the sample stage (means);



(Pixel) block image means for photographing the sample video and displaying the image ;



Superposition means (overlap means) for superimposing the electron lattice and the sample image

It is characterized by having.

In Example 1,

The index detection means for detecting the index of the sample stage is at least:

Indicator means drawn (marked) on the sample stage;

Index imaging means for imaging the index (CCD camera here),

(Pixel) block image means for converting the image from the index imaging means into a (pixel) block image

(Using capture means (A / D conversion means));

Known coordinate detection means for detecting the coordinates of the index shown in the (pixel) block image (the pixel in which the index is shown or a specific pixel or a specific block image in the block image) Position information and absolute position information)

Is provided.

The indicator means includes a light spot, a drawing point, a hole, a line, a surface, a grid (including a correction grid if necessary), etc., and these indexes are used to display a background image and an index in an image captured by the index imaging device. Any index can be used as long as it can be separated. The index may be color or monochrome. In the case of color, there is an advantage that a plurality of indicators can be easily separated and used. In the case of monochrome, it is easy to obtain high resolution.

The index may be an active index that radiates electromagnetic waves like a light spot, or may be a passive index that radiates nothing from itself, such as a drawing point. Passive indicators are universal and easy to maintain. In addition, active indicators are often resistant to disturbances and can carry special signals, for example, signals. For example, if the stage moves beyond a certain threshold, if the grid image is fixed, it moves greatly. It may be used as a relay that continuously measures the range, or may be used to control the operation by sending a signal every time.

Here, the points marked on the sample stage were used.

A CCD camera was used as the index imaging means . Any imaging means such as a CMOS, a vidicon, or a saticon can be used. Then, the (pixel) block image means converts the video into a (pixel) block image . As a specific example, a capture unit (A / D conversion unit) is used, and a (pixel) block image of 720 × 480 dots is used as an example described later.

The known coordinate detecting means for detecting the coordinates of the index reflected in the video of the (pixel) block image is a known (index) light spot tracking means (apparatus) for detecting the index (light spot) present in the camera video. ) Etc. to detect the coordinate value of the indicator.

At this time, as one block of the known light spot tracking means (device), 3 pixels or more of image pixels (image as final data after A / D conversion) in the index imaging means are set as one block, and at least the final convergence block In terms of the relationship between resolution and field of view, a configuration of preferably 4 blocks or 9 blocks is adopted, and a specific position coordinate for this block, here, as an example, the position of the center of gravity in the block is the position coordinate. Use as The coordinates were made to correspond to the lattice coordinates of the measurement range lattice of the electronic lattice described later.

At this time, as the (index) coordinates, the position coordinates of a pixel (image as final data after A / D conversion) converted by a known capture means (A / D conversion means) or a collective image of dots are adopted. . For example, in the case of 720 × 480 dot capture, the two-dimensional orthogonal coordinates by (x, y) are used, and the coordinates of the pixel from the origin (0, 0) to the highest point of (720, 480). At this time, the position of the center of gravity of the block was made to correspond to this coordinate value. Of course, it does not have to be the center of gravity, and as an example, it may be the coordinate value of any pixel constituting the block.

Here, the (pixel) block image means for displaying the sample video image is:

Microscope objective lens as an example (here, DIN standard 40 times, 10 times, 20 times, 100 times, 80 times, 60 times the Izure of the objective lens) (here, 1/3 inches CCD) and a CCD camera standards such as The sample image from the sampled image pickup means is an image that can be displayed on the display means.

Although non-standardized sample imaging means may be used, the standardized sample imaging means that is compatible is recommended and effective. Here, unless otherwise noted, a 40 × objective lens + 1/3 inch CCD is used.

Then, the image captured from the sample imaging means is captured by the (pixel) block image means using the same capture means (A / D conversion means), and the image captured (A / D conversion) is also an image of 720 × 480 dots. did. And

The field of view of the sample image was 125 μm × 92 μm, and a pixel pitch of about 0.174 μm × about 0.192 μm was adopted. (When using a DIN 40x objective lens + 1/3 inch CCD)

The field of view of the index image was adjusted with a zoom lens and set to 12,500 μm × 9200 μm. (Moving tolerance in one indicator)

This 12.5 mm × 9.2 mm is the range of the electron lattice. (It is also the visual field range of the index image.) Covers an area of a little over 18 mm × 18 mm of general-purpose cover glass.

Here, in order to widen this range, it is advisable to set indices at about three places. As an example, red, blue, and green indexes are marked with an interval of about 25 mm.

Here, the difference between the sample imaging means and the index imaging means is only the difference in the optical system such as a lens. Further, here, the sample image pickup means and the index image pickup means are installed such that their image pickup element surfaces are parallel to the same plane. Of course, any positional relationship may be used as long as the positional relationship between the two imaging means is known.

The electron lattice means generates the electron lattice .

Specifically, first, the lattice interval, that is, one side of the lattice was set to 50 μm here. Of course, a lattice of any size may be generated.

As an example, in the x direction, grid lines are sequentially drawn from the origin as Xn = n · {G / (F / Hr)} from the origin. G is the size of one side of the lattice, F is the field length, and Hr is the horizontal resolution. In the case of Yn, Hr is Vr (vertical resolution). n is a positive integer and represents the nth lattice line.

As an example, lines are drawn at intervals such as Xn = 50 / (125/720) = 288 (dots) by a known line segment drawing means in the storage means.

Similarly, in the y direction, Yn = n · {G / (F / Vr)} is taken as an example, and Yn = 50 / (92/480) = 261 (dots) is taken as an example. Draw with line segment drawing means.

As the storage means used here, the storage means having the same arrangement of pixels or block images having the same resolution as the sample image was used. Of course, the superimposing means (overlap means) may also be used, so that the image may be drawn on the same storage means as the sample image. In the case of different resolutions, image matching means or resolution matching means (resolution adjustment means) for matching the two may be used.

Here, the reference positions of the sample image and the index image are aligned in advance by aligning the reference position of the sample and the reference position of the index. As a specific example, when the origin (0, 0) is determined as shown in FIG. 2 and the index is positioned at the origin, the index or index imaging means is set to X− so that the center of the visual field is positioned at the left end of the slide glass. Move and adjust in the Y direction. This is determined at the time of design and may be manufactured at the origin determined as in the above example at the time of manufacture.

In this case, the storage means may be main storage, but may be drawing-like storage means such as an image board, or overlay storage means.

The superposing means (overlap means) superimposes the electron lattice on the sample image and displays it on the display means.

The overlapping means (overlap means)

The sample image from the (pixel) block image means and the electron lattice from the electron lattice means are superimposed. The superimposed video is stored in the storage means or displayed on the display means.

Here, the display means may use a display means with the same resolution, but may use a display means with a higher resolution. At that time, image matching means for matching the resolution of the display means, the resolution of the sample image, and the resolution of the electronic lattice image is used. These may be realized by a known GUI (graphic user interface). Further, the ratio of the sample image and the index image can be set constant even when the magnification is different or the same, but the ratio of the display image and the two images (the sample image and the index image) varies. There is a case. The superimposing means (overlap means) may be provided with (resolution) adjusting means for adjusting these.

At this time, when the sample stage is moved and the index coordinates of the index image are moved in the 1-dot x direction, the lattice of the sample image moves 100 dots in the x-direction. (The sample stage moves about 17 μm in the x direction.)

Similarly, when the index of the index image is shifted by 1 dot in the y direction, the lattice of the sample image moves 100 dots in the y direction. (Moves about 20 μm in the y direction.)

[Modification]

Of course, if the total number of pixels of the index video is increased, the grid moves more smoothly as the number is increased. In this example, the grid moves as smoothly as possible when it has about 10,000 times as many pixels as the sample imaging means. On the other hand, even if the magnification of the index image is increased, it moves smoothly, but if an image sensor with the same number of pixels is employed, the field of view (grid range) becomes narrower.

Here, this intermittent movement can be handled without increasing the number of pixels.

Electronic lattice correction means may be used.

As an example, the electronic lattice correction means includes a (correction) lattice or a (correction) line group having a slightly different pitch from the block or pixel pitch of the (pixel) block image constituting the index image. Prepare it again. (If it is a color imaging means, the index and the correction grid may be separated by changing the color.)

As an example, for a length of 10 pixels or 10 blocks in a (pixel) block image, nine correction grids or nine correction line groups (as an example) have a length of 9 pixels or 9 blocks (actual). 5 black, 4 white, and the background color at both ends are marked as a correction grid on the sample stage. Then, while one pixel or one block index moves, the video value from the correction grid sequentially changes to each pixel or each block of 10 pixels or 10 blocks of the (pixel) block image.

Under this condition, every time the index moves by one pixel (one block), that is, every time the index moves by about 1/10 pixel or about 1/10 block, the amplitude value of the image of the correction grid is 1 pixel or 1 The peak value moves for each block (when the center of the amplitude value of the pixel is 0, the-peak and the plus peak are alternately moved by one pixel or one block). If the movement amount of the peak value is used as the movement amount of the electronic lattice, a smooth movement amount of about 1/10 can be obtained. That is, one movement amount of the peak value generated between the correction grid and the pixel (block image) corresponds to about 1/10 of the movement amount of the index pixel under this condition, and the correction grid means moves this movement amount. Information is transmitted to the electronic lattice means to move the electronic lattice. Specifically, the amount of movement of the index pixel is 17.5 μm here, and the amount of movement when the correction grid means is used is 1.75 μm. It can be seen that the movement of the electron lattice is quite smooth.

Although the number of pixels is about 10 in the above example, there is no limitation on the number of lines of the correction grid. It goes without saying that the resolution of the correction grating may be increased to the limit where the S / N ratio is allowed.

Using this electronic lattice correcting means , the electronic lattice means (which can determine the shape and size of the electronic lattice) may connect the peak values on the storage means to draw a large lattice. This is an example in which the electronic lattice correction unit serves as a relative index detection unit and performs relative coordinate detection (position detection). That is, the relative position information of the electron lattice with respect to the sample image is generated.

As an example, in the case of the above condition, a single line appears in 10 pixels in the index image, and at this time, the difference in magnification between the index imaging unit and the sample imaging unit is 100 to 1, so that a 175 μm grid is included in the sample image. Therefore, if a 50 μm grid is instructed to the electronic grid means, a 50 μm grid is drawn on the sample image by multiplying by 50/175 = 0.286 (reduction magnification). (In this case, the relative position information is often provided.)

In this case, if (absolute) position information such as the grid number or grid coordinates is not required, an index for detecting the (absolute) position is not necessary. Therefore, the (absolute) position detection index is not marked on the sample stage. Also good.

Here, if position information is necessary, it may be determined in the same manner based on the index, or (absolute) position information may be obtained by using a correction grid whose spatial frequency changes as the correction grid. In this case, the electronic lattice correction means is a type of index detection means for detecting the (absolute) position.

Note that the y direction may be set in the same manner, but the same is true for square pixels and equally spaced sampling. If the pixel is not a square pixel (equal interval sampling), the aspect ratio may be corrected by the aspect ratio correcting means for correcting the aspect ratio, and the display means may be a square lattice.

Here, as the index coordinates, it is desirable to use coordinates obtained by correcting lens distortion by the index coordinate correcting means. This comprises at least known lens correction means .

[Operation]

As shown in FIG.

As an example, a longitudinal groove probe (FIG. 6), a continuous circle probe (FIG. 7), or a qualitative sampling No. 9 probe, which is a quantitative sampling means capable of quantitatively collecting a sample from a periodontal pocket, is used. Samples (plaque phase or plaque layer) or liquid components (liquid phase) are collected in a glass slide, diluted with water if necessary (it is always desirable to dilute at a constant volume), and covered if necessary. Press the sample with a glass and place it on the sample stage. Then, the sample is photographed by the sample imaging means.

And when you turn on the power,

The sample image is displayed on the display means as shown in the upper diagram of FIG.

When the electronic lattice is further turned ON, an electronic lattice having the same scale size as the sample image is electronically drawn on the video as shown in the lower diagram of FIG. 4 and FIG. If the length of the sample is 50 μm, it appears as the same length as one side of the grating.

When the sample stage is moved, the observation position of the sample is moved accordingly. At the same time, the index position moves in the same manner, and the electronic lattice moves in accordance with the above conditions. As an example, when moved to X μm and the X axis, the electron lattice moves to the X axis by X μm (intermittently depending on conditions). The same applies to the Y axis. This means that the surgeon observes the lattice and microorganisms in the sample in the same manner as the lattice is drawn on the cover glass or slide glass.

Then, manually count the number of white blood cells in the lattice and write it down, or the measurement range setting means automatically uses the known automatic number counting means within the lattice designated by the index coordinate values. May be counted. You can just look at it. If necessary, size, length, volume, etc. may be measured using an electronic grid.

[effect]

The size, length, shape, etc. of structures such as leukocytes can be measured (two-dimensional measurement or two-dimensional quantitative measurement).

That is, two-dimensional quantitative measurement and two-dimensional qualitative measurement can be performed.

Here, if the standardized sample imaging means such as the DIN 40 × objective lens + 1/3 inch CCD is used, the video is standardized, so that anyone can measure wherever it is compatible. This greatly reduces diagnostic errors between doctors.

Conventionally, a grid is inked on a slide glass or a cover glass. In this case, it is very troublesome because it is necessary to ink the slide glass and the cover glass one by one. Moreover, although it was very expensive, the conventional cheap slide glass and cover glass can be used by this invention. Furthermore, the shape and size of the lattice can be set to any shape and size. An example is a polygonal lattice of triangles or more.

[Modification]
In the above embodiment, the sample is not particularly adjusted.

Collect plaque in a slide glass and dilute with a certain amount of diluent, for example, raw food, phosphate buffer, (artificial) saliva, (artificial) tissue fluid or about 3 μl to 6 μl of water, for example, 3 μl of water (for example) It is desirable to always dilute with a constant amount.) And it is better to press the sample with a cover glass.

Further, when the sample is obtained by collecting the liquid phase (including fluid and semi-fluid components) in the gingival crevice with a continuous probe (FIG. 7) or the like, it may not be necessary to dilute. In that case, don't worry about the diluent.

Further, in the above embodiment, a general slide glass and cover glass are used, but a special slide glass and cover glass may be used.

A specific example is a slide glass with a constant depth as shown in FIG. Using this, the number of indices such as white blood cells per unit lattice of the lattice and the number of index objects per unit volume (the number of white blood cells) may be calculated based on this depth. (There is also an advantage that the pressure contact conditions are likely to be the same.)

In the above embodiment, an electronic grid is used. However, coordinate presenting means for displaying the coordinate position on the grid as shown in FIG. 8 may be provided to display the coordinates on the display image. In this example, the maximum value of the x coordinate value is 720 / 2.88 = 250, and the maximum value of the y coordinate value is 480 / 2.61 = 183. The coordinate position may be displayed anywhere on the grid, and the display character may use a transparent color or a non-transparent color. It can also be displayed anywhere on the screen, such as the corner of the screen, or on a second monitor.

In this case, when the coordinate presenting means is turned on, the coordinates of the current grid are shown. (As an example, the lower figure of FIG.

When the coordinate presenting means is turned off, the coordinate value disappears, and when the electronic lattice is turned off, the electronic lattice disappears and only the sample image is displayed. (Upper figure of Figure 4)

Quantification normalized microscope apparatus of the second embodiment is provided with at least three-dimensional quantitative measuring means.

[Second Embodiment]
The third embodiment is directed to embodiments of the three-dimensional quantitative measurement by quantification normalized microscope apparatus.

[Constitution]

In addition to the configuration of Example 1

The three-dimensional (quantitative) measuring means includes at least a depth means .

If the sample is diluted further,

A dilution rate means is provided.

If it is not diluted, it is not necessary.

Here, the depth means is

1 Depth means using a constant depth slide glass, a constant depth cover glass, a slide glass with a known depth, and a means for storing or inputting and storing a known depth by a cover glass with a known depth. Here, the slide glass whose depth is known includes a glass whose depth is known with respect to the lattice position, that is, the position can be specified even if the depth differs depending on the location.

2 In the case of depth means comprising at least a thickness meter for measuring the depth, that is, the thickness of the sample.



This is realized by any one of the above means 1 and 2 or a combination thereof.

In a specific example, if the depth is known by using the constant depth cover glass or the like as an example of the case of the depth means 1, the depth value is input to the depth means. Enter in A specific example is 15 μm or the like.

The sides of the lattice are known from the electronic lattice means. Here, it is 50 μm as an example. Here, it is better to follow the statistical population estimation to determine how many index objects are measured.

Therefore, the number of white blood cells per unit volume is known.

[Operation]

As with the apparatus of the first embodiment, the power is turned on, the same operation as that of the first embodiment is performed, and the following operation is further added. That is, as a specific example of additional operation

In FIG.

The number of white blood cells in one lattice (50 μ × 50 μm) is 8.

Next, the depth is input to the depth means using a keyboard or the like. Here, 15 μm is input.

Here, when the sample is diluted, the dilution rate means calculates the dilution rate, but here, it is not diluted and 1 is input or the dilution rate means is not used.

As a result, the number per unit volume, that is, the density is calculated and displayed on the display means. Ie

In the above example with the depth value from the depth means and the grid dimensions from the electron grid means, the density is

The density means calculates 2.13 × 10 ⁻⁴ (pieces / μm ³ ), and the display means

2.13 × 10 ⁵ / mm ³

And so on.

Further, when the white blood cells in the blood are measured, the normal value is 4,000 to 8,000 / mm ³ , and thus 1.5 to 3 × 10 ⁻¹ in a hexahedron having a grid of 50 μm and a depth of 15 μm. You will see it. This means that 1.5 to 3 white blood cells are observed when viewing 10 fields of view. Needless to say, it is necessary to perform a statistical sample for this number. (Adopting population estimation means described later is desirable.) In this way, the density of white blood cells in blood can also be measured.

Needless to say, if the sample is not diluted, the sample can be quantified without using the dilution rate means. In particular, the plaque phase (layer) often needs to be diluted, and the liquid phase often needs to be diluted particularly when the collection fee is small.

[effect]

These mean that three-dimensional quantitative measurement and three-dimensional qualitative measurement can be performed.

That is, as a specific example, the number of white blood cells per unit volume is known. Similarly, the number of microorganisms, structures, tissues, etc. per unit volume is known.

In addition, the amount collected by the No. 9 probe for qualitative sampling can be measured, and as a result, quantitative measurement can be performed.

[Modification]

The depth means was a cover glass with constant depth, but various thickness gauges (laser, capacitance, differential transformer, etc.) were used to measure the thickness of the sample for automatic or manual input. Depth means may be used.

As an example of the case of the depth means 2, it is possible to use a cover glass and a slide glass that are the cheapest and versatile extremely general cover glasses and general and extremely general slide glasses.

As a specific example,

For example, the cover glass is placed on a slide glass, the thickness is measured by the thickness measuring means, the thickness is measured with the sample sandwiched, and the difference is set to the thickness. In this case, when the thickness measuring means is connected to a computer by USB or the like, the depth is automatically stored in the depth means. In this case, the thickness is measured for each sample.

Further, when the thickness measuring means is installed on the sample stage or the like, the thickness can be measured on the electronic lattice, so that measurement data (matrix) of the electronic lattice and the thickness can be used. In this case, the depth is accurately measured even if there is a thickness variation in the surface of the cover glass due to “” bending ”or“ deflection ”of the cover glass. This can be quantified only with a slide glass with a constant depth, but it is effective even when a slide glass with a constant depth is used, and the accuracy further increases.

These are also examples of three-dimensional quantitative measurement means.

[General modification]
Add or modify any of the above embodiments or combinations thereof,

You may provide the number display means which displays a number on an electronic lattice. A number (here, coordinate value) is associated with the block, the number is stored in the storage means, and displayed or hidden by a switch of the number display means.

When the number display means is turned on, a number is displayed for each grid. (Fig. 8)

You may employ | adopt the length display means which displays a length value on an electronic lattice. A numerical value corresponding to a length value for each lattice or the scale is stored in the storage means, and the length value is displayed by a switch of the length display means.

When the length display means is turned on, the length value (length scale) is displayed, and the length of the sample can be measured.

Here, it is possible to provide a random extraction means for randomly displaying numbers from the number display means for displaying numbers on the electronic lattice and sampling (measuring) each number. (FIG. 8) At this time, the measurement value is transmitted to the measurement range setting means corresponding to the grid number to be measured and stored.

Furthermore, a population estimation means for displaying one or a combination of a histogram, an average, a deviation, a sampling number, and a reliability each time the random sampling is performed may be provided. This is reasonable because the minimum necessary sampling can be recognized and the sampling (measurement) can be stopped at a minimum. (Fig. 8)

The position of the sample stage may be the position information of the electronic grid using existing position detecting means such as a magnescale, an operating transformer, or a stepping motor capable of detecting the position. In that case, most of the present invention is more expensive than the present invention, and the mechanism is likely to be complicated and large, but a substitute can be made.

Further, the position information of the electron lattice may be obtained by irradiating the sample with a laser, integrating the speckle speed, and using a position detecting means by laser speckle for detecting the position. In this case, an index, an index imaging unit, and an index detection unit are unnecessary.

The first embodiment of the present invention is an example excluding the means for aligning the electron grid by the position detection (means).

A sampling and quantification means may be used in the embodiment.

In particular,

1. Continuous circle probe (continuous donut-shaped probe) (Figure 7)

2 Vertical groove probe (Fig. 6)

3 Dropper for periodontal pocket

Or any combination of these.

1, the liquid component (fluid, semi-fluid component) in the periodontal pocket, ie, the liquid phase, the biozone of the inner epithelium (tissue phase or tissue layer), the submarine plaque (plaque layer or plaque phase) of the root surface Since the sample can be quantitatively collected by any one or a combination thereof, the aforementioned diagnosis can be quantitatively performed. In addition, samples in the deep part and the shallow part from the pocket bottom to the pocket opening can be sampled intermittently by sampling only the sample in the circle, or continuous on the surface of the circle continuous with the circle. Can be sampled continuously.

2, the liquid component (including fluid component and semi-fluid component) in the periodontal pocket, that is, the liquid phase, the biozone of the inner epithelium (tissue phase or tissue layer), the submarine plaque (plaque layer or Since the sample can be quantitatively collected in any one of the plaque phases) or a combination thereof, the aforementioned diagnosis can be quantitatively performed. Further, it is possible to continuously sample a deep sample and a shallow sample from the pocket bottom to the pocket opening.

3 can be used to quantitatively collect liquid components (including fluid components and semi-fluid components) in the periodontal pocket, that is, the liquid phase, so that the aforementioned diagnosis can be quantitatively performed. Moreover, the liquid phase of a specific part from the pocket bottom to the pocket opening can be spot-collected. In continuous suction, continuous samples can be collected, and in intermittent suction, intermittent samples can be collected.

As an example of standardized sample imaging means ,

When the image transmission means is digital, that is, when a signal obtained by directly A / D converting each pixel of the CCD is used, a CCD using a CCD pixel (pitch) of about 7 μm dots for a DIN standard objective lens 40 times is used. You may normalize by a pixel (pitch). Also,

At least when the video transmission means uses an existing analog video signal, the DIN standard objective lens 40 times may use a standardized sample imaging means standardized by a final sampling pixel to be described later. A standard with a 3 inch CCD and a DIN standard objective lens 40 times may be used. The two may have the same standard with a width. Furthermore, when standardization is performed with sampling pixels that take into account all of the CCD pixels, capture sampling conditions, video signal processing timing, and the like, the following is performed.

Here, when an image having a 0.175 μm pixel pitch (when the CCD pixel is about 7 μm and using a 40 × DIN standard lens) is used as a condition for one pixel in the captured image, when the 480 iDV standard is adopted under this pixel condition, In the case of a 40 × DIN standard objective lens, the field of view is about 126 μm × 84 μm. Furthermore, when it goes to 720 × 512 of the PAL standard, it is about 126 μm × 90 μm. Incidentally, this microscope field of view (40 × DIN standard objective lens) employs a pixel pitch of 0.174 μm × 0.19 μm at 125 μm × 92 μm. The reason why the vertical pixels are slightly larger is that they are limited to the size of the CCD pixels, the number of CCD pixels, 480 lines of the 480iDV standard (capture board sampling conditions), or the NTSC standard. Further, it may be a standard at least in the horizontal direction (x direction). Of course, it goes without saying that the pixel pitch of the CCD satisfying the above conditions, the bandwidth of the transmission system, and the like are necessary.

Here, as a more detailed standard than this standard, the output signal from the CCD camera may be used as a standardized sample imaging means in which the output signal from the CCD camera is equivalent to the high-vision or DVD standard 480i by S-Video (or higher) output or digital output. good. In this case, diagnosis errors between doctors are reduced on a global scale.

The standard analog phase contrast microscope standard seems to be optimal for a 1/3 inch CCD camera with S-Video output for a finite focal length objective lens with a DIN standard of 40 times. The horizontal resolution is 480 or more, 512 or more, and 525 is the most desirable. (It is preferable that the camera has a resolution of 720 × 480 or more. If possible, 720 × 512 or more and 525 are most desirable.)

Here, it is considered that the capture board is optimally sampled at 720 × 480 to 720 × 525 pixels. Of course, there are many conditions that can ensure a higher resolution and a wider field of view, but the standard that can handle the most widely used video standard is the 40 × DIN objective lens + 1/3 CCD + S-Video + 720 × 480 capture standard. . Therefore, it is valuable because most people can diagnose and exchange opinions with the same image.

Needless to say, in the future, it will be desirable to develop standards with higher resolution and higher field of view.

The drawing image of the electronic lattice means is a quadrangular shape, but it may be a polygonal or irregular lattice.

A lattice having a specific shape matching the shape of the object to be measured may be used. As an example, many white blood cells infiltrated into the gingival sulcus are round, so a round lattice may be used.

In this case, since it can be set to the shape of the index object, sampling with less error can be performed.

The quantification normalized microscope apparatus, what sample may be applied to the index.

The embodiment or modification of the present invention may be manufactured entirely by hardware, or a portion replaceable by software may be replaced.

The embodiments or modifications of the present invention may be implemented singly or in combination. Moreover, you may use for another use. In addition, the above means may be configured alone or in any combination, such as an operator or manufacturer selecting, using, and manufacturing.

As an example, the image recognition unit (image recognition means deformation is one example of a quantitative measuring means in quantifying standardized microscope apparatus is the invention, at least the image size, the correlation means for obtaining correlation by the form And the like, and further, by combining fluorescent means or spectroscopic means.
The image recognition means here is a kind of quantitative measurement means. That is, quantification normalized microscope apparatus is the present invention comprises at least the image recognition means (a quantitative measurement means), further fluorescent means, spectroscopic means, characterized by incorporating the properties means such as a motion analysis unit Therefore, the observation object such as microorganisms in the sample image can be specified by the size, shape, and properties (spectral characteristics, fluorescence characteristics, movement characteristics, etc.) of the observation objects such as microorganisms.

In a specific example of this combination,
Prebotella Intermedia size is measured in the form of short bacillus,
In mature strains, the length is about 1.7 μm to about 2.3 μm (about 1 μm when immature)
Width from around 3.5μm to around 4.6μm (around 1μ when immature)
Correlation means for calculating the correlation with a short filter of the above-mentioned size, etc. and fluorescence means for exciting the sample at an ultraviolet wavelength of about 300 nm to less than 400 nm and displaying only visible light in the sample image With. Here, Prevoterra Intermedia emits red light (fluorescence).
And the sample image A sample taken from the oral cavity by the quantification normalized microscope apparatus in such the collection dosing means, here, further samples were excited radiation at said wavelength, sample the image Let it be image B. Then, the image recognition means superimposes the short gonococcus region which is correlated with the correlation coefficient by the size bacterium (region or shape) on the image B, and then superimposes the image B on the image B. The image recognizing means selectively displays the short gonococcus region of the image A by the red filter processing means. (Selection display is, for example, surrounding an image that matches a condition with a specific shape with a surrounding line, coloring it, etc. Here, the specific shape is a short bacillus shape with the above size. Here, the image to be used can be selected only by B. However, since it is easier to see the image A for separation from the background image or the like, the image A and the image B are used here.
Then, it is possible to see images of only Prevoterra intermedia with a considerable probability.
At this time, the image-recognizing means switches the phase contrast microscopic image, or superimposes the fluorescent image on the phase contrast microscopic image, extracts it from the background image, and observes the red portion of the short bacilli that has been confirmed. Also good. Furthermore, the intensity of this red light emission intensity may be measured and used as the Prevoterra Intermedia activity index.

As another example, may be combined incorporate an image recognition means and separating means to quantify normalized microscope device is present invention. The image recognition means is a kind of quantitative measurement means. (The image recognition means includes correlation means for obtaining a correlation based on at least the size and shape of the image.)
Here, the wavelength used for the spectroscopic means is about 1038 cm-1 or about 1033 cm-1, etc., and the absorption image B (same processing as the Prevoterra intermedia), Streptococcus mutans and size (1 μm) are absorbed only at this wavelength. Sphere), and the correlation of the image A in the shape (a chain shape of about 1 μm sphere and an interval of about 0.5 μm or a streptococcus shape in the form of a bead), the image recognition means, as in the case of the Prevotella intermedia, For example, A (phase contrast microscope image) is marked or selected. Then, the marked or selected portion is a Streptococcus mutans with a considerable probability. The image may be used only for the image B as in the Prebotella intermedia, but it is desirable to use the image A together.
Further, another Streptococcus genus may be selected and displayed using about 1018 cm−1 which is the wavelength of another Streptococcus genus. That is,
In the case of Streptococcus sarivarius, about 1048 cm-1,
In the case of Streptococcus sanguisu, about 1018cm-1
May be used to identify each. Also,
In the case of a bimodal peak such as Streptococcus mites, it may be separated by extracting about 1038 cm-1 and about 1024 cm-1 equally. If the 1024cm-1 peak is equivalent to the 1038cm-1 peak, the Streptococcus mitisses is identified, the 1038cm-1 peak is identified as red, the 1024cm-1 peak is identified as blue, the yellow is Streptococcus mitithus, and the red is Streptococcus mutans. It may be identified.
In addition, since there may be a change depending on the amount of water that comes from the sample holding condition, the change due to moisture may be corrected by a moisture correction unit using a known calibration unit.

Here, the motile microorganism may be extracted by a specific exercise by the exercise analysis means.

As an example, the moving speed is analyzed by a known speed analyzing means, and the image recognizing means analyzes the shape and size of the motile microorganism and marks or selectively displays the motile microorganism.

Of course, in the quantitative measurement of the genus Prevotella or Streptococcus, it may be used as a second quantitative measurement together with the measurement range lattice, the constant depth cover glass means, or the like to measure the density.

As an example used for other purposes, the position of the sample stage can be detected by the index detection means, and this may be used as an object marker. As another example, the measurement object may be used for any substance such as a metal structure, an organic compound, and an inorganic compound instead of a microorganism.

Since the sample can be quantified and standardized, accurate inspection, observation, examination, etc. can be performed in the microscopic examination. And diagnosis and treatment based on it.

Block diagram an example of quantification normalized microscope apparatus. Operation An example of quantification normalized microscope apparatus. Block diagram an example of such depth means was adopted quantified normalized microscope apparatus. An example of a video by quantification normalized microscope apparatus. An example of a slide glass with a constant depth. An example of a flute probe as a sampling quantitative means. An example of a continuous circle probe (continuous donut-shaped probe) as a sampling and quantification means. An example image quantitation normalized difference microscope apparatus.

Claims (7)

Quantification standardized microscope equipment
An XY stage means for holding a sample collected from a living body and moving it at least in the XY direction in order to observe each part of the sample;

Sample imaging means for imaging sample image information from the sample, and a sample (pixel) block image means for obtaining a two-dimensional image block composed of pixels consisting of resolution units from the sample imaging means and the XY stage means Index image capturing means for capturing the imaged index, and index (pixel) block image means for obtaining a two-dimensional image block composed of pixels comprising resolution units from the index image capturing means;


A correction grid having a constant pitch installed on the XY stage means;
In the index (pixel) block image obtained by imaging the correction grid,
By taking a slightly different number of correction grids as a video image with respect to the number of blocks in the block image, the image pickup pixel pitch and the index (pixel) block image pitch are slightly shifted, resulting in the beat obtained by the shift amount. High-resolution relative position information obtained by capturing image amplitude
Or
(Absolute) position information from the index image obtained in the same manner as described above using a pitch change correction grid having a spatial frequency change as the correction grid
Use one or a combination of
At least to improve positioning resolution or / and
At least by the electron lattice correction means to improve the resolution of the electron lattice,

The both (pixel) block images are
Based on the amplitude interval of the beat image in the x direction of the X stage means obtained by the electronic lattice correction means from the correction grid imaged on the index pixel block, the electronic grid means performs a grid interval on the sample pixel block. The interval setting
A mechanism based on relative positioning between the two pixel blocks obtained by setting in the same way for the y direction in the Y stage ;

A mechanism constituted by absolute positioning of the two pixel blocks obtained from (absolute) position information from the index image obtained in the same manner using the pitch change correction grid.
In a mechanism comprising either or a combination of


At least one point index installed on the XY stage means;

The coordinates of the point index are (index) a coordinate detection means that uses either the center of gravity of the final convergence block of the light spot tracking means (device) or individual pixels.

A quantified standardized microscope device characterized by comprising.
Quantification standardized microscope equipment
An XY stage means for holding a sample collected from a living body and moving it at least in the XY direction in order to observe each part of the sample;

Sample imaging means for imaging sample image information from the sample, and a sample (pixel) block image means for obtaining a two-dimensional image block composed of pixels consisting of resolution units from the sample imaging means and the XY stage means Index image capturing means for capturing the imaged index, and index (pixel) block image means for obtaining a two-dimensional image block composed of pixels comprising resolution units from the index image capturing means;


A correction grid having a constant pitch installed on the XY stage means;
In the index (pixel) block image obtained by imaging the correction grid,
By taking a slightly different number of correction grids as a video image with respect to the number of blocks in the block image, the image pickup pixel pitch and the index (pixel) block image pitch are slightly shifted, resulting in the beat obtained by the shift amount. High-resolution relative position information obtained by capturing image amplitude
Or
(Absolute) position information from the index image obtained in the same manner as described above using a pitch change correction grid having a spatial frequency change as the correction grid
Use one or a combination of
At least to improve positioning resolution or / and
At least by the electron lattice correction means to improve the resolution of the electron lattice,

The both (pixel) block images are
Based on the amplitude interval of the beat image in the x direction of the X stage means obtained by the electronic lattice correction means from the correction grid imaged on the index pixel block, the electronic grid means performs a grid interval on the sample pixel block. The interval setting
A mechanism based on relative positioning between the two pixel blocks obtained by setting in the same way for the y direction in the Y stage ;

A mechanism constituted by absolute positioning of the two pixel blocks obtained from (absolute) position information from the index image obtained in the same manner using the pitch change correction grid.
In a mechanism comprising either or a combination of


At least one point index installed on the XY stage means;

A coordinate detection means that uses the coordinates of the point index as the (index) center of gravity of the final convergence block of the light spot tracking means (device) or individual pixels;

An electron grid means for generating an electron grid based on the index position information from the coordinate detection means means;
A quantified standardized microscope device characterized by comprising.
Quantification standardized microscope equipment
An XY stage means for holding a sample collected from a living body and moving it at least in the XY direction in order to observe each part of the sample;

Sample imaging means for imaging sample image information from the sample, and a sample (pixel) block image means for obtaining a two-dimensional image block composed of pixels consisting of resolution units from the sample imaging means and the XY stage means Index image capturing means for capturing the imaged index, and index (pixel) block image means for obtaining a two-dimensional image block composed of pixels comprising resolution units from the index image capturing means;


A correction grid having a constant pitch installed on the XY stage means;
In the index (pixel) block image obtained by imaging the correction grid,
By taking a slightly different number of correction grids as a video image with respect to the number of blocks in the block image, the image pickup pixel pitch and the index (pixel) block image pitch are slightly shifted, resulting in the beat obtained by the shift amount. High-resolution relative position information obtained by capturing image amplitude
Or
(Absolute) position information from the index image obtained in the same manner as described above using a pitch change correction grid having a spatial frequency change as the correction grid
Use one or a combination of
At least to improve positioning resolution or / and
At least by the electron lattice correction means to improve the resolution of the electron lattice,

The both (pixel) block images are
Based on the amplitude interval of the beat image in the x direction of the X stage means obtained by the electronic lattice correction means from the correction grid imaged on the index pixel block, the electronic grid means performs a grid interval on the sample pixel block. The interval setting
A mechanism based on relative positioning between the two pixel blocks obtained by setting in the same way for the y direction in the Y stage ;

A mechanism constituted by absolute positioning of the two pixel blocks obtained from (absolute) position information from the index image obtained in the same manner using the pitch change correction grid.
In a mechanism comprising either or a combination of


At least one point index installed on the XY stage means;

A coordinate detection means that uses the coordinates of the point index as the (index) center of gravity of the final convergence block of the light spot tracking means (device) or individual pixels;

An electron grid means for generating an electron grid based on index position information from the coordinate detection means means;

Overlaying means (overlap means) for superimposing the electron lattice and the sample image,
A quantified standardized microscope device characterized by comprising.
Quantification standardized microscope equipment
An XY stage means for holding a sample collected from a living body and moving it at least in the XY direction in order to observe each part of the sample;

Sample imaging means for imaging sample image information from the sample, and a sample (pixel) block image means for obtaining a two-dimensional image block composed of pixels consisting of resolution units from the sample imaging means and the XY stage means Index image capturing means for capturing the imaged index, and index (pixel) block image means for obtaining a two-dimensional image block composed of pixels comprising resolution units from the index image capturing means;


A correction grid having a constant pitch installed on the XY stage means;
In the index (pixel) block image obtained by imaging the correction grid,
By taking a slightly different number of correction grids as a video image with respect to the number of blocks in the block image, the image pickup pixel pitch and the index (pixel) block image pitch are slightly shifted, resulting in the beat obtained by the shift amount. High-resolution relative position information obtained by capturing image amplitude
Or
(Absolute) position information from the index image obtained in the same manner as described above using a pitch change correction grid having a spatial frequency change as the correction grid
Use one or a combination of
At least to improve positioning resolution or / and
At least by the electron lattice correction means to improve the resolution of the electron lattice,

The both (pixel) block images are
Based on the amplitude interval of the beat image in the x direction of the X stage means obtained by the electronic lattice correction means from the correction grid imaged on the index pixel block, the electronic grid means performs a grid interval on the sample pixel block. The interval setting
A mechanism based on relative positioning between the two pixel blocks obtained by setting in the same way for the y direction in the Y stage ;

A mechanism constituted by absolute positioning of the two pixel blocks obtained from (absolute) position information from the index image obtained in the same manner using the pitch change correction grid.
In a mechanism comprising either or a combination of


A correction grating having a constant pitch installed in the XY stage means;
In the index (pixel) block image obtained by imaging the correction grid,
By taking a slightly different number of correction grids as a video image with respect to the number of blocks in the block image, the image pickup pixel pitch and the index (pixel) block image pitch are slightly shifted, resulting in the beat obtained by the shift amount. High-resolution relative position information obtained by capturing the amplitude of the image, or
Using any one or a combination of (absolute) position information from the index image obtained in the same manner as described above using a pitch change correction grid having a spatial frequency change as the correction grid ,
At least to improve positioning resolution or / and
An electron lattice correction means for improving the resolution of at least the electron lattice,
A quantified standardized microscope device characterized by comprising.
Quantification standardized microscope equipment
An XY stage means for holding a sample collected from a living body and moving it at least in the XY direction in order to observe each part of the sample;

Sample imaging means for imaging sample image information from the sample, and a sample (pixel) block image means for obtaining a two-dimensional image block composed of pixels consisting of resolution units from the sample imaging means and the XY stage means Index image capturing means for capturing the imaged index, and index (pixel) block image means for obtaining a two-dimensional image block composed of pixels comprising resolution units from the index image capturing means;


A correction grid having a constant pitch installed on the XY stage means;
In the index (pixel) block image obtained by imaging the correction grid,
By taking a slightly different number of correction grids as a video image with respect to the number of blocks in the block image, the image pickup pixel pitch and the index (pixel) block image pitch are slightly shifted, resulting in the beat obtained by the shift amount. High-resolution relative position information obtained by capturing image amplitude
Or
(Absolute) position information from the index image obtained in the same manner as described above using a pitch change correction grid having a spatial frequency change as the correction grid
Use one or a combination of
At least to improve positioning resolution or / and
At least by the electron lattice correction means to improve the resolution of the electron lattice,

The both (pixel) block images are
Based on the amplitude interval of the beat image in the x direction of the X stage means obtained by the electronic lattice correction means from the correction grid imaged on the index pixel block, the electronic grid means performs a grid interval on the sample pixel block. The interval setting
A mechanism based on relative positioning between the two pixel blocks obtained by setting in the same way for the y direction in the Y stage ;

A mechanism constituted by absolute positioning of the two pixel blocks obtained from (absolute) position information from the index image obtained in the same manner using the pitch change correction grid.
In a mechanism comprising either or a combination of


A correction grid having a constant pitch installed in the XY stage means and at least one point index installed in the XY stage means;

Coordinate detection means for detecting index coordinates based on the point index;

Absolute position information obtained by the index;
Or
In the index (pixel) block image obtained by imaging the correction grid, the number of correction grids slightly different from the number of blocks in the block image is captured as a video, whereby the imaging pixel pitch and the index (pixel) block image are captured. High-resolution relative position information obtained by capturing the amplitude of the beat image obtained by the amount of the deviation due to the slight deviation of the pitch,
Or
(Absolute) position information from the index image obtained in the same manner as described above using a pitch change correction grid having a spatial frequency change in the correction grid;
Use one or a combination of
At least to improve positioning resolution or / and
An electron lattice correction means for improving the resolution of at least the electron lattice,
A quantified standardized microscope device characterized by comprising.
The quantified standardized microscope apparatus according to any one of claims 1 to 5,
A quantification standardized microscope apparatus comprising a continuous probe as a collection and quantification means for quantifying a sample collected from a living body or the like.
The quantified standardized microscope apparatus according to any one of claims 1 to 6,
A quantification standardized microscope apparatus comprising an image recognition means for recognizing a specific bacterium.