JPH03194451A - Cell analyzing apparatus - Google Patents

Abstract

PURPOSE:To form a cytogram which is simple in operation and is exact by automatically regulating the level of a detection signal to an optimum level just before measurement operations are carried out for each of the samples. CONSTITUTION:A regulating circuit 15 has a gain control circuit, an averaging circuit 18 and a comparing and regulating circuit 19. The gain control circuit can control the value of the amplification gain for the signal pulses to be inputted thereto. The average signal level value of the inputted signal pulses is obtd. in the circuit 18. The circuit 19 compares the average signal level value and the set value which is preset as an optimum range. This circuit 19 applies the feedback signal to the gain control circuit to allow the average signal level value to satisfy the set value. The regulating means 15 regulates the output signal of a photoelectric converting means 11 to regulate the output of the circuit 18 to an adequate signal level just before the actual measurement operation is made. The operation is simplified in this way and the exact cytogram is formed.

Description

[Detailed Description of the Invention] [Industrial Application Field] The present invention relates to a cell analysis device, and particularly to a device that analyzes cells based on the intensity of scattered light and fluorescence generated by irradiating individual cells with light. The present invention relates to a cell analyzer used in fields such as clinical testing and cell engineering in blood cell analysis.

[Conventional technology]

When analyzing blood cells, the intensity of the fluorescent signal obtained varies greatly depending on the type of sample, individual differences, etiology, and the type of fluorescent dye or monoclonal antibody used as pretreatment for analysis. arise.

Therefore, in conventional blood cell analysis, before the actual measurement, measurements are performed using the same sample to check the fluorescence intensity level, and when the signal level is too high, the amplifier gain is reduced manually to reduce the signal level. When the gain is too small, it is necessary to increase the gain and adjust the signal level to a predetermined level. after that,
Measure the sample again to obtain the original data.

Further, as a conventional adjustment device for a cell analyzer, there is one disclosed in Japanese Patent Application Laid-Open No. 63-63942.

Before the actual measurement, this adjustment device flows fluorescently labeled test particles, and the operator adjusts the voltage of the photomultiplier tube so that the signal given by the test particles is displayed as a predetermined histogram. Configured for repeated adjustment.

[Problem to be solved by the invention]

According to the conventional cell analyzer, the operator must check the signal strength and manually adjust the gain of the amplifier before the actual measurement. Normally, this manual adjustment takes a considerable amount of time, and its adjustment ability also depends on the experience of the measurer. In some cases, it is necessary to perform the adjustment work by performing measurements many times, which reduces work efficiency. There was a problem that the value decreased.

In addition, when measuring the same sample before the actual measurement to check the signal strength, when the amount of sample is small, valuable samples cannot be wasted and adjustments are difficult. A problem arises.

The purpose of the present invention is to eliminate the complexity of conventional equipment in which the gain is manually adjusted every time the conditions of the sample to be measured change, and to automatically adjust the signal strength to an appropriate predetermined level in a short time. The object of the present invention is to provide a cell analysis device that is simple and can produce accurate cytodialums.

[Means to solve the problem]

A first cell analyzer according to the present invention includes a flow cell including a detection region through which cells to be measured pass one by one;
An optical means for irradiating cells with light in a detection area, a photoelectric conversion means for detecting scattered light or fluorescence generated by the light irradiation and converting it into an electrical signal, and an optical means for analyzing the electrical signal and displaying and analyzing the analysis information. In a cell analyzer equipped with a data processing means for storing, an averaging means inputs an electrical signal to calculate the average intensity of scattered light or fluorescence, and an output of the averaging means immediately before actual measurement. and adjustment means for automatically adjusting the electrical signal output by the photoelectric conversion means so that the signal has an appropriate signal level.

A second cell analysis device according to the present invention is characterized in that, in the first configuration, the amplification means is configured to freely change the gain, and the adjustment means automatically adjusts the gain of the amplification means. shall be.

A third cell analysis device according to the present invention is characterized in that, in the first configuration, the photoelectric conversion means includes a control section that freely changes the conversion efficiency, and the adjustment means automatically adjusts the control section. do.

In the fourth cell analyzer according to the present invention, in the first configuration, the optical means has a control section that freely changes the output of the light source, and the adjustment means automatically adjusts the control section. Features.

A fifth cell analyzer according to the present invention has a trigger signal generation means in any one of the first to fourth configurations, and after the adjustment means has made adjustment for each sample, the trigger signal generation means The data processing means is characterized in that the data processing means is activated by a trigger signal outputted by the data processing means.

[Effect]

In the first to fourth cell analyzers according to the present invention, the averaging means and the adjusting means automatically adjust the circuit gain in a short time so that the signal level output from the photoelectric conversion means satisfies an appropriate required level. The adjusted signal is then used to perform actual measurements.

The fifth cell analyzer according to the present invention is configured to include a trigger signal generating means and to start the data processing means to perform the original measurement after the signal level adjustment by the adjusting means or the like is completed.

〔Example〕

Embodiments of the present invention will be described below with reference to the accompanying drawings.

FIG. 1 shows the configuration of a first embodiment of a cell analysis device according to the present invention. Reference numeral 1 denotes a flow cell, and the flow cell 1 has a wall portion made of a transparent member, and has a detection region therein through which cells to be measured are passed one by one.

A sheath flow is formed in the flow cell 1 by the diluted and stained blood sample and the sheath liquid, and the detection area is irradiated with light supplied from the light source 2. The type and number of cells are determined from several types of optical information obtained by this light irradiation, and the cells are analyzed.

The light irradiated from the light source 2 is condensed by the condenser lens 3 onto the detection area located in the center of the flow cell 1, and is focused on the sample. Scattered light is generated from cells passing through the detection area, and fluorescence is generated if the cells are stained with a fluorescent dye. Among the scattered lights, forward scattered light L1, which provides size information, and side scattered light L2 in a 90-degree direction, which provides internal information of the cell, play important roles in cell analysis. The forward scattered light L1 is collected by a condensing lens 4 and detected by a detector 8. Further, the side scattered light L2 and the fluorescence are collected by a condensing lens 14, branched by a quick clock mirror 12.13, and given to the respective detectors 9, 10.11.

The light separated by the dichroic mirror 12.13 passes through the filter 5.6.7, which passes only the light of the wavelength to be detected, and is then sent to each detector 9. 10. 11. These filters are selected to have optimal transmission characteristics based on the wavelength of the light source used and the fluorescence wavelength characteristics of the fluorescent dye used. Detector 8.9, 10.11
A photoelectric conversion element having a function of converting an optical signal into an electrical signal is used, such as a photodiode or a photomultiplier tube that can change the gain of the signal level. Each detector8. 9. 10. The electrical signals outputted from the control circuit 11 are input to the adjustment circuit 15 as mutually independent signal channels. The adjustment circuit 15 adjusts the gain of the signal level so that the signal input from each of the detectors has an optimal level, as will be described later.

Each signal optimally adjusted by the adjustment circuit 15 is processed by signal analysis and
The information is supplied to the display unit 16, where a cytogram is created and displayed or the number of cells is counted, and necessary information is obtained.

Next, the structure and operation of the adjustment circuit 15 will be explained in detail with reference to FIGS. 2 and 3. Figure 2 shows the adjustment circuit 15.
2 shows a signal processing configuration for one signal channel in FIG. The adjustment circuit 15 includes a gain control circuit 17 capable of changing the amplification gain for the input signal pulse and controlling the value of the gain from the outside, and an average signal level value of the input signal pulse. and the gain control circuit 1 which compares the average signal level value with a setting value set in advance as an optimum range, and controls the average signal level value to satisfy the setting value.
7 and a comparison/adjustment circuit 19 that provides a feedback signal 21 for adjusting the gain thereof, and these circuits are connected in series. The number of cells used for averaging in the averaging circuit 18 is about 20% of the total (for example, 1,600 to 2,000 cells). Further, the optimum range for setting the set value in the comparison/adjustment circuit 19 is determined by the range in which the cytogram can be written. The signal whose level has been appropriately adjusted by the action of the comparison/adjustment circuit 19 etc. is sent to the gain control circuit 1.
After being output from 7, it is supplied to the next stage signal analysis/display section 16.

The adjustment circuit section having the above configuration is connected to the detector 8°9.1.
0.11 signal channels are provided.

The operation of the adjustment circuit 15 of the cell analyzer having the above configuration is shown in a flowchart as shown in FIG. In the configuration of FIG. 2, the configuration of the adjustment circuit is shown in terms of hardware, but the functions of the adjustment circuit 15 can also be configured in terms of software according to the flowchart in FIG.

When cells to be measured begin to pass through the detection region of the flow cell 1, measurement and adjustment operations are performed to adjust the open signal level for an initial fixed period (for example, 2 to 3 seconds).

First, a signal due to scattered light or fluorescence is detected by each of the detectors 8.
9. 10. 11 (step 30). Next, each of the detected signals is averaged in the averaging circuit 18 as described above (step 31). In judgment step 32, the set value set by the comparison/adjustment circuit 19 is compared with the average value. If the average value is larger than the set value and the amplifier output signal is saturated,
When it deviates from the cytogram display, a feedback signal 20 is output for adjusting the gain of the gain control circuit 17 to reduce the signal level (step 33). When the average value is smaller than the set value and the distribution of cells cannot be distinguished on the cytogram, a feedback signal 20 is outputted to adjust the gain of the gain control circuit 17 to increase the signal level (step 34).
). When the average value becomes appropriate in relation to the set value, or when it is appropriate from the beginning, the gain value is memorized and retained (
Step 35). When the above processing is completed, the start signal for the original measurement is output in the next step 36,
A measurement operation is started (step 37).

When the adjustment measurement described above is completed, a trigger signal for starting the operation of the signal analysis/display section 16 that performs data processing is generated by a trigger generation circuit (not shown) (step 36), and the actual measurement is thereby started. will be started. During the above operation, the blood sample flows through the flow cell 1 at a constant flow rate. Further, the adjusted gain value is stored and further sent to the signal analysis/display section 16. In this way, the gain adjustment amount is provided as one of the parameters and can be used as an index of differences depending on the cell sample.

As described above, the system is configured to automatically adjust the signal level to the optimum level in a short time before performing the actual measurement for each sample, so it is possible to create the optimal cytogram for any blood sample. Accurate measurement results can be obtained.

Here, the reason why gain adjustment is required for the signal as described above will be explained.

There are various methods for fluorescent staining of blood cells, and different fluorescent dyes are used depending on the type of blood cells to be analyzed and the purpose. An example of staining a blood sample with a fluorescent dye will be described. By utilizing the specificity of staining using a fluorescent dye, it is possible to classify white blood cells into five categories using acridine orange, count reticulocytes using auramine O, etc. In addition, monoclonal antibodies labeled with fluorescent dyes such as FITC (fluorescein iso-thiacinet) and PE (phycoerythrin) can be used in immunology fields other than clinical testing, such as lymphocyte subset analysis and functional tests. Research related to engineering fields can also be conducted. However, the intensity emitted by these fluorescent dyes when they bind to the target binding site of blood cells varies depending on the type of dye, the state of the sample, etc., and is not always the same fluorescence intensity. Therefore, when measuring blood samples stained with different dyes, it is necessary to adjust the signal level each time. Due to these circumstances, the present invention allows the operator to omit the work of checking the signal level and adjusting the gain every time a different staining method is used.
The automatic adjustment allows efficient and accurate data to be obtained.

Next, a modified example of the adjustment circuit will be described based on FIGS. 4 and 5.

When the detector (for example, detector 9) is formed of a photomultiplier tube, its output has a characteristic that it is proportional to the applied voltage. Therefore, as shown in FIG. 4, an applied voltage control section 21 is provided for the detector 9,
The applied voltage outputted by the applied voltage control section 21 is configured to be controlled by the feedback signal 20 from the adjustment circuit 15'. With this configuration as well, effects similar to those of the above embodiment can be obtained. Note that the adjustment circuit 15' has basically the same circuit configuration as shown in FIG. 2, but does not include the gain control circuit 17, and instead simply includes a constant gain amplifier.

In the modified embodiment shown in FIG. 5, a light source output control section 22 is provided in the laser light source that is the light source 2, and the output of this light source output control section 22 is controlled by the feedback signal 20 output from the adjustment circuit 15'. This controls the laser output of the light source 2. This makes it possible to adjust the signal level of the fluorescence output, which is proportional to the amount of laser light. This configuration can also produce the same effects as those of the embodiments described above.

〔Effect of the invention〕

As is clear from the above explanation, according to the present invention, it is possible to automate the work of adjusting the level of the detection signal to the optimum level immediately before performing the actual measurement work for each sample. It is possible to omit extensive manual adjustments, significantly reducing the workload, making effective use of small volumes of precious samples, and furthermore, due to automation, the time required is extremely short compared to conventional manual adjustment. This has the effect that adjustment work can be carried out with

[Brief explanation of drawings]

FIG. 1 is a block diagram showing the overall configuration of the cell analysis device according to the present invention, FIG. 2 is a detailed circuit diagram of the adjustment circuit, FIG. 3 is a flowchart showing the automatic adjustment operation according to the present invention, and FIGS. The figure is a partial circuit diagram showing a modified embodiment. [Explanation of symbols] 1φ・@−・Fist・Flow cell 2・・・・・Light source 8.9, 10.11・・・Detector 15.15′・・Adjustment circuit 16・・...Signal analysis/display section 17...Gain control circuit 18...Averaging circuit 19...Comparison/adjustment circuit 21... ... Applied voltage control section 22 ...... Light source output control section

Claims (5)

[Claims] (1) A flow cell comprising a detection region through which cells to be measured pass one by one, an optical means for irradiating the cells with light in the detection region, and a method for detecting scattered light or fluorescence generated by the irradiation of the light. A cell analyzer is equipped with a photoelectric conversion means for detecting and converting into an electric signal, and a data processing means for analyzing the electric signal and displaying and storing analysis information. averaging means for calculating an average magnitude of intensity; and immediately before actual measurement, automatically adjusting the electrical signal output by the photoelectric conversion means so that the signal output from the averaging means has an appropriate signal level. A cell analysis device characterized by having an adjusting means for adjusting. (2) The cell analysis device according to claim 1, further comprising an amplification means configured to freely change the gain, and wherein the adjustment means automatically adjusts the gain of the amplification means. . (3) The cell analysis device according to claim 1, wherein the photoelectric conversion means includes a control section that freely changes conversion efficiency;
A cell analysis device characterized in that the adjustment means automatically adjusts the control section. (4) The cell analysis device according to claim 1, wherein the optical means has a control section that freely changes the output of the light source;
A cell analysis device characterized in that the adjustment means automatically adjusts the control section. (5) The cell analyzer according to any one of claims 1 to 4, further comprising a trigger signal generating means, and after the adjusting means performs the adjustment for each sample, the trigger signal generating means A cell analysis device characterized in that the data processing means is activated by an output trigger signal.