JPH0560752A - Cell analyzer - Google Patents

Abstract

PURPOSE:To achieve automatic analysis and to enhance the efficiency of analysis and the accuracy of data by determining the best smoothing technique by smoothing data and comparing results. CONSTITUTION:A window is manually set to start novel measurement, and specimen cells linealy flow on the center axis of a sheath liquid and that of flow cell 10. The specimen cells are irradiated with a laser beam L and the scattered light from the cells are detected by photodetectors 15a-15d and the intensities I0, I90, Ig, Ir of forward scattered light, 90$0 scattered light, green fluorescence and red fluorescence are analized by a CPU 22. That is, when the number of cells in the window reaches a predetermined number during measurement, the measurement is completed and the operation in the window, the unification of the number of cells and the analysis of fluorescence characteristics are performed. When there is a specimen to be measured further, the final fraction due to just-before measurement is used as a window to perform measurement in the same way as in the previous measurement and, after the completion of measurement, automatic trigger operation is performed. That is, a plurality of smoothing operations are applied to the obtained data of the intensities I0, I90 and the respective calculated fractions are subjected to fraction evaluating operation to determine the best smoothing method. After determination, the unification of the number of cells is performed in the same way as the previous measurement. Therefore, the optimum smoothing processing is always applied and cell analysis is performed.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a cell analyzer to which flow cytometry is applied. More specifically, it smoothes data in the process of processing optical information of cells and compares smoothed results. The present invention relates to a cell analyzer which enables to determine the best smoothing method.

[0002]

2. Description of the Related Art Flow cytometry is a method in which cells or particles labeled with a fluorescent dye, for example, are made to flow in a fine liquid flow, and a laser beam is emitted to each cell flowing in a row due to a hydrodynamic focusing effect. The cell is analyzed by irradiating the cells and instantaneously measuring the intensity of scattered light or fluorescence generated by the cells, that is, cell light information. This flow cytometry is
It has the characteristic of being able to analyze a large amount of cells at high speed and with high precision, and is widely used in clinical tests to research.

As a cell analyzer using the above flow cytometry, a flow cell for forming a fine liquid flow, a light source (for example, a laser) for irradiating cells flowing in the flow cell with a light beam, and this light beam are used. It is known that a photodetector that detects cell light information from irradiated cells and converts it into an electric signal and a computer that performs analysis processing of the cell light information converted into the electric signal and the like are known.

In this conventional cell analyzer, a cell suspension (sample) labeled with a fluorescent dye is made to flow in a flow cell together with a sheath liquid. A sheath flow is formed in the flow cell, and cells flow in a line on the central axis of the flow cell due to the hydrodynamic focusing effect. The intensity of scattered light and fluorescence generated by irradiating these cells with a light beam is detected by a photodetector such as a photomultiplier tube as a parameter constituting cell light information.

Now, when a cell suspension contains a plurality of cell populations, it may be desired to analyze one of these cell populations. For example, in the case of the analysis of lymphocyte subsets in human blood, a sample obtained by reacting blood with a monoclonal antibody labeled with a fluorescent dye and hemolyzed is used. , Monocytes, and granulocytes are included, and it becomes necessary to select lymphocytes from the cell light information obtained by the measurement.

Therefore, the cell light information of the necessary cell population is manipulated.
Automatically select and collect without relying on the author's subjectivity
Cell analysis equipment with a function that allows
For example, Japanese Patent Application No. 2-247 related to the applicant's prior application
I have 17. This cell analyzer has multiple parameters.
Cell light information obtained from
Sets the analysis area based on two or more parameters.
Collect the cell light information of the target cell population in the analysis area of.
For example, in the case of lymphocyte analysis, two-dimensional smoothing
After that, as shown in FIGS. 10 (a) and 10 (b), 90 °
Scattered light intensity I₉₀, Forward scattered light intensity I₀The histogram of
Create each. I ₉₀The minimum point p than the histogram of
₁, P₂, P₃But I₀The minimum point p than the histogram of
_Four, P_FiveIs extracted.

From these minimum points p _{1 to} p ₅ , I ₉₀ -I
_{The 0} cytogram is fractionated as shown by the broken line in FIG. In FIG. 8, b shows the distribution of lymphocytes, c shows the distribution of monocytes, and d shows the distribution of granulocytes. Also,
Although a represents a distribution of debris, debris is composed of a membrane component of red blood cells and the like, and is usually removed in the stage of noise processing.

Since the distribution B of lymphocytes is included in the fraction B shown by the solid line in FIG. 8, the cell light information is collected by using this fraction B as the analysis region. In this way, the collected cell light information is further analyzed for fluorescence characteristics and subjected to processing such as determination of positive rate. According to the applicant of the present invention, Japanese Patent Application No. 63-1993033,
Japanese Patent Application No. 63-193072 has already been applied,
Either method is used as is. Furthermore, Japanese Patent Application Sho 6
In 3-320915, the applicant of the present application discloses a method for unifying the number of cells belonging to a set analysis region, but this method is used as it is.

Further, in the cell analyzer of Japanese Patent Application No. 24-24717, the analysis area closer to the distribution shape of the target cell population is set and the number of cells is unified, so that the reliability of the measurement result is improved. Therefore, it is possible to further promote automation and efficiency of analysis.

[0010]

The cell analyzer equipped with the auto-trigger function has advantages that the measurement is certainly automated and that objective data processing can be performed. However, when the number of cells is small or when the distribution of cells is uneven or biased, a histogram of the forward scattered light intensity I ₀ or a histogram of the 90 ° scattered light intensity I ₉₀ from the I ₉₀ -I ₀ cytogram is created. If it is not possible or even if a histogram can be created, a so-called "tooth gap" may occur in the quantization of the continuous number.
In this case, it becomes impossible to extract a local minimum point or a point having a predetermined threshold value from each histogram, so that auto-trigger calculation cannot be performed properly, automation of measurement is hindered, and reliability of data is impaired. Sometimes. In particular, when the measurement target is sparsely distributed over a wide range such as granulocytes, or when it is a blood sample or the like of a patient to which an anticancer agent has been administered, the above problems may be remarkably observed.

The present invention has been made in view of the above, and is further improved by determining the best smoothing method by comparing data smoothing and the result thereof with the processing of the cell light information processing means. It is an object of the present invention to provide a cell analysis device which improves the accuracy of data by automating and improving the efficiency of analysis.

[0012]

In order to achieve the above object, the cell analyzer of the present invention has the constitutions listed in the following (a) to (i). That is, (a) a flow cell in which a cell suspension or the like is flown, and (b) a light source for irradiating a light beam to the cell suspension or the like flowing in the flow cell,
(C) Cell light information detecting means for detecting cell light information consisting of a plurality of parameters for each particulate component of cells or the like contained in the cell suspension or the like irradiated with this light beam, and (d) this cell Analysis area setting means for setting an analysis area based on one or more parameters obtained by the optical information detecting means, and (e) a target cell population within the analysis area set by the analysis area setting means. Cell light information collecting means for collecting cell information of (6), (f) cell light information detected by the cell light information detecting means and cell light information of the target cell population collected by the cell light information collecting means In a cell analysis device comprising cell light information processing means and (g) output means for outputting a processing result of the cell light information processing means, in the cell light information processing means (f),
(H) a plurality of smoothing means for smoothing the cell light information detected by the cell light information detecting means and the cell light information of the target cell population collected by the cell light information collecting means;
For each of the processing results by the plurality of smoothing means,
Smoothing determining means for performing an evaluation operation for determining the quality of the smoothing processing and determining an optimum smoothing means according to the operation result.

[0013]

The smoothing means performs a plurality of smoothing processes on the cell light information detected by the cell light information detecting means and the cell light information of the target cell population collected by the cell light information collecting means. The smoothing determination means performs an evaluation operation on the individual data smoothed by the plurality of smoothing means for evaluation of the quality of the smoothing processing, and determines the optimum smoothing means according to the operation result. ..

That is, according to the present invention, the optimum smoothing process is always performed, and the cell light information after the process is
Cell analysis will be performed.

[0015]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS An embodiment of the present invention will be described below with reference to FIGS. FIG. 3 is a diagram showing the configuration of the cell analyzer of the example. 2 is an auto sampler,
The sample rack 2a is loaded with a plurality of sample containers 3 ,. The sample rack 2a is driven by a drive mechanism (not shown) to position a designated sample directly below the sample suction tube 4. Furthermore, this auto sampler 2
Is equipped with a shaking mechanism and a cooling mechanism (not shown), and shakes the sample at regular intervals and keeps the samples in the loaded sample containers 3, ... 3 at a low temperature (for example, 4 ° C to 10 ° C). To do.

The sample suction tube 4 is a three-way valve 1
Connected to one port. Sample feeding tubes 6a and 6b are respectively connected to the other two ports of the three-way valve 5, and the sample feeding tube 6a and the sample suction tube 4 or the sample feeding tubes 6a and 6b are connected to each other. You can A sample pump 7 is provided at the other end of the sample liquid feeding tube 6a. On the other hand, the other end of the sample liquid feeding tube 6b is opened inside the sheath liquid feeding tube 8.

The sheath liquid feeding tube 8 has one end connected to the liquid feeding pump 9 and the other end connected to the flow cell 10. The flow cell 10 is made of quartz glass or the like, a sheath flow is formed in the flow channel 10a inside, and cells or particles in the sample are lined up on the central axis of the flow channel 10a due to the hydrodynamic focusing effect. Shed

The liquid flowing out of the flow cell 10 is guided to the waste liquid tube 11 and stored in the waste liquid tank 12.
The cell analyzer is equipped with a sheath liquid tank (not shown), and the sample pump 7 and the liquid feed pump 9 are replenished with the sheath liquid. Further, the liquid feeding system including the autosampler 2, the pumps 7, 9 and the like can be hermetically sealed to prevent biohazard.

Around the flow cell 10, a laser (light source) 14, a photodetector (cell light information detecting means) 15a, 1
5b, 15c and 15d are arranged. The laser beam L from the laser 14 is applied to cells (or particles) flowing through the flow channel 10a. Signal light is generated from the cells (or particles), but the light in the forward direction is condensed as the forward scattered light by the lens 16a and enters the photodetector 15a. A beam blocker 17 prevents the laser beam L from directly entering the photodetector 15a.

On the other hand, the signal light in the 90 ° direction from the cell is
It is condensed by the lens 16b. A part of this signal light is reflected by the dichroic mirror 18a and enters the photodetector 15b for detecting 90 ° scattered light. A part of the signal light transmitted through the dichroic mirror 18a is further reflected by another dichroic mirror 18b, transmitted through the filter 19a, and the photodetector 15c for green fluorescence is emitted.
Is received by. The light transmitted through the dichroic mirror 18b passes through the filter 19b and is received by the photodetector 15d for red fluorescence. For example, the laser 14 is an argon laser or a helium neon laser, the photodetector 15a for the forward scattered light is a photodiode, and the other detector 1 is used.
A photomultiplier tube is applied to 5b, 15c, and 15d.

Photodetectors 15a, 15b, 15c, 15d
The received light signal of is amplified by the signal processing circuit section 20 to remove noise, and then the analog / digital (A / D) converter 2
It is converted into a digital signal by 1 and taken into the MPU 22. The MPU 22 roughly has a function related to auto-trigger, a function related to cell number counting / unification, and other functions. As a function relating to the auto-trigger, for example, if the method of Japanese Patent Application No. 63-193033 is applied, a function of creating a cytogram and a histogram of each of the forward scattered light intensity I ₀ and the 90 ° scattered light intensity I ₉₀ , The function of extracting the minimal points from these histograms and fractionating the cell light information into fractions, the function of smoothing the entire cytogram or within the fraction, and the final fraction within the fraction containing the target cell population It has a function to decide.

The functions related to counting and unifying cell numbers include a function of storing the final fraction of the immediately preceding measurement, a function of counting the cell number using the final fraction of the immediately preceding measurement at the time of this measurement, and a function obtained at the time of this measurement. It has a function of adjusting the number of cells in the final fraction to a specified number n ₀ . Other functions include a function of performing fluorescence analysis on optical information of a target cell population in which the number of cells is uniform, a function of controlling the sample pump 7, the liquid sending pump 9, and the autosampler 2.

The MPU 22 includes a floppy disk drive 23, a keyboard 24, a CRT 25 and a printer 26.
Are connected. The floppy disk drive 23 is
This is for saving measurement conditions (protocol), measurement data, etc. on a floppy disk. Keyboard 24
MP for protocol selection / setting or other commands
It is for inputting into U22. The CRT 25 is for monitoring the measurement, and the printer 26 is M
It is for printing out the processing result of the PU 22, for example, a cytogram or a histogram.

Next, the operation of the cell analyzer of the embodiment is shown in FIG.
It will be described with reference to the flowchart shown in FIG. First,
Samples that have been processed according to the purpose of analysis are placed in the sample containers 3 and loaded into the sample rack 2a. For example, a sample for the analysis of lymphocyte subsets is fluorescein isothiocyanate (FITC) in the blood of patients.
It is obtained by reacting the OKT4 monoclonal antibody labeled with and the OKT8 monoclonal antibody labeled with phycoerythrin (PE), followed by hemolysis.

When the cell analyzer is powered on, the RO
A program is read into MPU22 from M (not shown), and the system starts up [Step (hereinafter referred to as ST)]
1, see FIG. 2]. Next, a protocol suitable for the sample to be measured is selected and set. This protocol includes measurement conditions such as detection gains of the photodetectors 15a, 15b, 15c and 15d and correction calculation. This is because the method of treating the sample differs depending on the purpose of measurement, and, for example, the reactivity of the monoclonal antibody used in each treatment with the cells differs, so that the photodetectors 15a, 15b,
It is necessary to change the detection gains of 15c and 15d, and since the binding modes of each monoclonal antibody and the fluorescent dye are different, the correction calculation also needs to be changed accordingly.

Next, in ST2, it is determined whether or not a command to perform an auto trigger has been input from the keyboard 24. If this determination is YES, the process branches to ST3,
If NO, the process branches to ST4. Further, in ST3, it is determined whether or not a command that the current measurement is performed according to a new protocol different from the previous measurement is input.
22 determines. If the determination is NO, the process branches to ST8, and if the determination is YES, the process branches to ST4.

Now, the description will proceed assuming that the process branches from ST2 or ST3 to ST4. In ST4, the operator sets the window B on the I ₉₀ -I ₀ cytogram using the keyboard 24 (see FIG. 5A). When the window B is set, the measurement is started (ST5). In ST5, the sample rack 2a is driven, and the sample container 3 containing the sample to be measured first is positioned directly below the sample suction tube 4. Then, the three-way valve 5 is switched so as to connect the sample suction tube 4 and the sample solution sending tube 6a,
The sample suction tube 4 descends into the sample container 3, the sample pump 7 is driven to the suction side, and the sample is transferred to the sample solution feeding tube 6
a is aspirated.

Next, the three-way valve 5 is switched so as to connect the sample solution sending tubes 6a and 6b, the sample pump 7 is driven to the solution sending side, and the sample is sent to the solution sending tube 8. On the other hand, the liquid sending pump 9 is driven to the liquid sending side, and the sheath liquid is sent to the flow cell 10. In the flow channel 10a,
A sheath flow is formed, and cells flow in a line on the central axis of the flow channel 10a due to the hydrodynamic focusing effect. The laser beam L is irradiated to each of the cells, and the forward scattered light intensity I ₀ , the 90 ° scattered light intensity I ₉₀ , the green fluorescence intensity I _g , and the red fluorescence intensity I _r are measured. The cell light information is sequentially stored in a floppy disk or the like as needed.

During the measurement, the number of cells belonging to the window B is counted. In ST6, it is determined whether or not the number of cells has reached a predetermined n, and if YES, ST7.
If NO, ST6 is repeated. The predetermined value n is
For a given cell number n ₀ are larger n ₀ + alpha (see FIG. 7). In ST7, the measurement is completed, and the statistical calculation and various parameters of the cell light information in the previously set window B are performed.

At this time, the number N of cells in the window B becomes smaller than the above-mentioned predetermined number n of cells, but since it is counted with a margin of α from the beginning, it will not fall below n ₀ . .. In ST12, a uniform cell number process is performed. This processing is performed by using a method similar to, for example, Japanese Patent Application No. 24-24717.

For the data in which the cell numbers are unified in ST12, the fluorescence characteristic analysis is further performed (ST13). In this fluorescence characteristic analysis, for example, a histogram is created for each of the green fluorescence intensity I _g and the red fluorescence intensity I _r , and the positive rate is calculated. The result of this fluorescence characteristic analysis is
It is displayed on the CRT 25 (ST14) and printed out from the printer 26 (ST15).

At ST16, it is judged whether or not there is a sample to be further measured. If this determination is YES, ST
Return to 1 and if NO, end analysis. Well, ST
Returning to step 1, if the same protocol as the previous measurement is used and auto trigger is performed, go through ST2, ST3, and then ST8.
Go to processing. In ST8, the final fraction B obtained in the immediately preceding measurement is set again as a window on the I ₉₀ -I ₀ cytogram (see FIG. 5 (b)). Then, the measurement is started in exactly the same manner as ST5, the number of cells in the final fraction B is counted, and it is determined whether or not this has reached a predetermined number n (ST
10).

When the determination in ST10 is YES, ST1
The process proceeds to step 1 to end the measurement and perform auto trigger calculation. In this auto-trigger calculation, I ₀ , I
_A plurality of smoothing operations are separately applied to ₉₀ data (ST11
1, see FIG. 1). The plurality of smoothing operations are, for example, 2
Dimension smoothing only, point sequence smoothing only, 2
It refers to three types of combined processing of the dimensional smoothing method and the point sequence smoothing method, all of which are performed separately. Here, the two-dimensional smoothing method is performed by the method disclosed in Japanese Patent Application No. 24-24717. The masking of the two-dimensional smoothing method is shown in FIG.
It is performed like. On the other hand, the point sequence smoothing method uses three points (window boundary points) of point P _n , point P _{n + 1} , and point P _{n + 2} as shown in FIG. 11, and point P _{n + 1} is point P _n and point P _n. It is processed by determining how uneven the line segment between P _{n + 2} is. That is, when there is unevenness of a predetermined value or more, smoothing is performed by adopting the line segment between the point O and the point P _{n + 1} and the intersection point P ′ of the line segment between the point P _n and the point P _{n + 2.} Going forward (above ST11
1).

Next, the I ₉₀ -I ₀ cytogram, I ₉₀ , and I ₀ histograms are created for the smoothed I ₉₀ and I ₀ data (ST112). Then, for each histogram obtained by the three smoothing operation determines whether it is possible to extract the minimum point p ₁ ~p ₅ (ST11
3). Here, if there is even one minimal point that cannot be extracted, the process proceeds to ST115, and if the minimal points can be extracted for all, the process proceeds to ST114 and subsequent steps (ST113).

In ST114, the minimum point p of the histogram is₁
~ P_FiveFraction B based on (see FIGS. 10 (a) and (b))
₁Is set (see FIG. 5 (b)), and this fraction B₁Inside
Collect data. And this fraction B₁About the special application
63-193030 (see FIGS. 9 (a) and 9 (b))
Fractions that have been processed (ST114) and then calculated
B ₁′ ′ Fraction evaluation calculation is performed (ST116).

This operation is performed on all three types of data after the smoothing operation, and the smoothing operation considered to be the best is determined based on the result. Specifically, the fraction evaluation calculation is performed by the aspect ratio (F ₂ −) of the target cell population in the fraction.
_{F 1) / (R 2 -R} 1) or the eccentricity (X ₀ -R ₁₎
/ (R ₂ -R ₁₎ and _{(Y 0 -F 1) / (} F 2 -F 1)
Is a calculation for obtaining the value of (see FIG. 5B). The point (X ₀ , Y ₀ ) is the maximum position of the cell appearance frequency in the final fraction B ₁ ′.

Then, the result of the above-described fraction evaluation calculation for the smoothed data which seems to be the best is compared with the value input in advance, and it is judged whether or not the processing up to this point is normal ( ST117). If it is determined to be normal,
The best considered smoothing method described above is employed to finally determine fraction B ₁ '. On the contrary, if it is determined to be abnormal, the process moves to ST115 and an alarm is issued (ST11
5).

The same applies when the local minimum point cannot be extracted in the determination of ST113, and the operator is notified by displaying that fact on the CRT 25 (ST115).
The fraction is manually set by the operator (ST11
8). Predetermined statistical calculations and parameter calculations are also performed for the fractions thus set (ST119). S
At T120, it is determined whether to end the fraction determination.
For example, if there is an input from the keyboard 24 to redo the final fraction B ₁ ′, this determination is NO and ST11 is again entered.
Return to 8. Otherwise, the determination is yes and the process returns to the main routine of FIG.

Alternatively, the automatic trigger calculation in ST11
Is the minimum point p as in ST7. ₁~ P_FiveExtract
And fraction B₁After setting, this fraction B₁Within I₉₀,
I₀Apply the selected smoothing process to the data to obtain the final fraction
B₁'May be determined. In each case, the cell number is
Since it is counted by the last fraction B immediately before,
B₁Number of cells in ‘₁Is different from n, but n₀Below
There is no such thing. Therefore, in this case as well, the number of cells is unified as before.
Processing is performed (ST12). And ST13 to ST15
Is processed.

FIGS. 6 (a) and 6 (b) show granule spheres.
The final fraction B is calculated₁’
Showing I₉₀-I₀In the cytogram, Fig. 6 (a) shows the secondary
If the original smoothing process is not performed, FIG.
The case where the dimension smoothing processing is performed is shown. First
In Fig. 6 (a), the final fraction B₁’
The final fraction B₁’May not be set properly.
It is high. On the other hand, in FIG. 6 (b),
Final fraction B₁'Boundary is smooth and the final fraction B ₁'Is suitable
You can confirm that it is set to off.

Further, in FIGS. 12 (a) and 12 (b), the auto-trigger calculation is similarly performed for granulocytes.
FIG. 12A shows the case without the point sequence smoothing method, and FIG. 12B shows the case with the point sequence smoothing method.
In FIG. 12 (a), there are places where irregularities are remarkably generated.
In FIG. 2 (b), it can be confirmed that the boundary of the final fraction B ₁ 'has become slightly smoother and the remarkable unevenness has been improved.

As described above, in the cell analyzer of the embodiment, the data can be appropriately smoothed within the fraction or the entire I ₉₀ -I ₀ cytogram, so that it is difficult to extract a minimum point or a small number of data. Even in such a case, the auto-trigger calculation can be appropriately performed, automation of measurement of the calculation result can be promoted, and reliability can be improved. Although lymphocytes and granulocytes have been described in the above examples, the present invention is applicable to a wide range of sample analysis of leukocytes such as monocytes, erythrocytes, and cultured cells.

Further, the auto-trigger system is not limited to that of Japanese Patent Application No. 63-1903033, but Japanese Patent Application No.
No. 3-193072 or Japanese Patent Application No. 62-22884.
It is possible to change the number and other contour tracking methods as appropriate.

[0044]

As described above, in the cell analyzer of the present invention, the cell light information processing means is the cell light information detected by the cell light information detecting means and the target cell population collected by the cell light information means. The plurality of smoothing means for smoothing the cell light information and the cell light information obtained by the smoothing means are calculated by an arithmetic expression for evaluation, and the smoothing means for determining the optimum smoothing means based on the calculation result. And a deciding means for deciding the change.

Therefore, not only a situation in which a histogram cannot be created or a "missing tooth" is generated in the histogram, but also a situation in which the original data is significantly separated by the smoothing process can be avoided. .. In other words, even if the number of cells is small or the distribution of cells is uneven or biased (for example, the distribution of granulocytes in blood), the auto-trigger operation can be appropriately performed, and the results can be evaluated at the same time. It has an advantage that not only the automation but also the reliability of the measurement result can be further improved.

[Brief description of drawings]

FIG. 1 is a flowchart illustrating an auto-trigger calculation of a cell analyzer according to an embodiment of the present invention.

FIG. 2 is a flowchart illustrating the entire operation of the same cell analyzer as in FIG.

FIG. 3 is a block diagram illustrating the configuration of the same cell analyzer as in FIG.

FIG. 4 is a diagram illustrating a two-dimensional smoothing process of the cell analyzer.

FIG. 5 is a cytogram of 90 ° scattered light intensity-forward scattered light intensity for explaining the auto-trigger calculation of the cell analyzer.

FIG. 6 is a 90 ° scattered light intensity-forward scattered light intensity cytogram comparing the case where two-dimensional smoothing was not performed for granulocytes.

FIG. 7 is a diagram illustrating a cell number unification process of the cell analyzer.

FIG. 8 is a cytogram of 90 ° scattered light intensity-forward scattered light intensity for explaining the auto-trigger of the conventional cell analyzer.

FIG. 9 is a cytogram of 90 ° scattered light intensity-forward scattered light intensity for explaining the auto-trigger of the conventional cell analyzer.

FIG. 10 is a histogram of 90 ° scattered light intensity and forward scattered light intensity for explaining the auto trigger of the cell analyzer.

FIG. 11 is a diagram illustrating a point sequence smoothing method process.

FIG. 12 is a diagram showing cytogram data of 90 ° scattered light intensity-forward scattered light intensity when the smoothing process is not performed and when the point sequence smoothing process is performed.

[Explanation of symbols]

 ST111 Plural smoothing means ST113 to ST117 Smoothing determining means

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[Procedure amendment]

[Submission date] August 18, 1992

[Procedure Amendment 1]

[Document name to be amended] Statement

[Correction target item name] 0024

[Correction method] Change

[Correction content]

Next, the operation of the cell analyzer of the embodiment is shown in FIG.
It will be described with reference to the flowchart shown in FIG. First,
Samples that have been processed according to the purpose of analysis are placed in the sample containers 3 and loaded into the sample rack 2a. For example, as a sample for analyzing lymphocyte subsets, patient blood is reacted with OKT4 monoclonal antibody labeled with fluorescein isothiocyanate (FITC) as green fluorescence and OKT8 monoclonal antibody labeled with phycoerythrin (PE) as red fluorescence. And then hemolyzed.

Claims (1)

[Claims] 1. A flow cell in which a cell suspension or the like is flown, a light source for irradiating the cell suspension or the like flowing in the flow cell with a light beam, and cells or the like contained in the cell suspension or the like irradiated with the light beam. Cell light information detecting means for detecting cell light information consisting of a plurality of parameters for each of the particulate components, and analysis for setting an analysis region based on one or more parameters obtained by the cell light information detecting means Area setting means, cell light information collecting means for collecting cell information of a target cell population in the analysis area set by the analysis area setting means, and cell light information detected by the cell light information detecting means And cell light information processing means for processing the cell light information of the target cell population collected by the cell light information collecting means, and output means for outputting the processing result of the cell light information processing means. In the cell analysis device, the cell light information processing means smoothes the cell light information detected by the cell light information detecting means and the cell light information of the target cell population collected by the cell light information collecting means. For each of the plurality of smoothing means to be performed and the processing results of the plurality of smoothing means,
A cell analysis apparatus, comprising: a smoothing determination unit that performs an evaluation calculation for determining the quality of the smoothing process and determines an optimum smoothing unit based on the calculation result.