Patent ID: 12188857

DESCRIPTION OF THE EMBODIMENTS OF THE INVENTION

The first embodiment will be described below. However, the present invention is not limited to only the embodiment shown in this embodiment.

Flow Cytometer and External Server

The flow cytometer is installed in, for example, a hospital, a laboratory or the like, and is connected to the external server11via a network12such as the Internet as shown inFIG.1A. InFIG.1A, a plurality of flow cytometers10A to10C are connected to the external server11. The network12is a communication medium such as the Internet, a virtual private network (VPN), a wide area communication network (WAN), a public switched telephone network (PSTN) or the like, and is not limited insofar as communication between the external server11and the flow cytometer10is possible. Hereinafter, when there is no need to distinguish the flow cytometers10A to10C, the flow cytometer is simply referred to as “flow cytometer10”.

The external server11is installed in a database center or the like and has a storage device for storing the database D. The database D includes measurement conditions corresponding to information for specifying measurement items of the flow cytometer10. The flow cytometer10receives the measurement condition from the external server11via the network12. The database D of the external server11is configured to accept changes and additions of measurement conditions at any time from the server administrator. In a general-purpose flow cytometer such as the flow cytometer10, since it is possible to add new measurement items applicable to a reagent and even the same reagent at any time, it becomes possible to provide new measurement items and measurement conditions to the flow cytometer of the user facility by having the server administrator register the measurement condition/s corresponding to the new measurement item/s in the database D.

In this case, the measurement item is one or more items measured by the flow cytometer10, and is, for example, the type of the particle, the kind of the substance present in the particle and the like. Specifically, the type of cell, type of protein, kind of sugar chain, kind of lipid, type of glycoprotein, type of glycolipid, kind of lipoprotein, kind of nucleic acid, kind of biological component such as cylinder and the like can be mentioned. The information for specifying the measurement item is not limited insofar as the measurement item can be specified. For example, the measurement item may be the name of the measurement item itself, or the type of stain which stains the particle, the substrate of the enzyme, information on the reagent necessary for detecting particles such as antibodies, or the name of the antigen. The information for specifying the measurement item also includes identification information of the reagent mixed with the particles in order to measure the measurement item.

The number of flow cytometers10connected to the external server11is not limited, and a single flow cytometer10also may be connected to the external server11.

As shown inFIG.1A, the flow cytometer10includes a flow cytometer main body13, and an information processing device14connected to the flow cytometer main body13. As shown inFIG.1B, the flow cytometer main body13is provided with a suction unit710configured to be able to move up and down and move horizontally. The user positions the sample container containing a manually prepared measurement sample at the position21outside the flow cytometer main body13to measure the measurement sample in the sample container. The suction unit710moves just above the position21and the measurement sample is suctioned from the sample container positioned at the position21.

Optical System of Flow Cytometer

FIG.2is a brief diagram showing the optical system of the flow cytometer10according to one embodiment. The flow cytometer10includes a flow cell20through which a particle-containing liquid containing particles in a sample passes, light sources101and124for irradiating light on particles passing through the flow cell20, and light receiving elements100A to100F for optically detecting optical information on light derived from the particles and outputting a detection signal converted into an electric signal.

Here, the sample is a suspension of particles suctioned by the flow cytometer. The particles may be artificial particles such as magnetic beads or plastic beads. The particles also may be biological components such as cylinders, and also may be microorganisms, animal cells, plant cells and the like.

It is preferable that the particles emit one or more lights when irradiated with predetermined light. Light emitted from particles when irradiated with predetermined light is collectively referred to as light derived from particles. Light derived from the particles includes scattered light and luminescence. The light derived from the particles may be light of any wavelength, but preferably is light having a peak wavelength in the range of 400 nm to 850 nm. More specifically, the light derived from the particles is preferably fluorescence. The light derived from the particles also may be light emitted by a substance contained in the particles themselves. Alternatively, light derived from the particles may be labeled with a luminescent substance such as a fluorescent substance, and light emitted from the luminescent substance may be detected as light derived from the particles. The peak wavelength of light derived from particles is preferably different for each measurement item.

The particle-containing liquid is a liquid containing the particle suspension liquid suctioned from the sample into the flow cytometer, and includes a diluting liquid as necessary.

The optical information is information included in one or two or more light wavelength spectra emitted from particles. The light wavelength spectra include the individual light wavelengths and light wavelength regions included in a light wavelength spectrum and the respective light wavelengths or the intensities of the light wavelength regions. Individual light wavelengths and wavelength ranges can be specified by which one of the one or more light receiving elements described later has received light. Each light wavelength or intensity of each light wavelength region can be specified by an electric signal output from the light receiving elements100A to100F which have received light.

The case where light derived from particles is scattered light and fluorescence will be specifically described hereinafter as an example.

The light emitted from the light source101irradiates the flow cell20via the collimator lens102, the dichroic mirror103, and the condenser lens104. The forward scattered light of the light derived from the particles passing through the flow cell20is collected by the condenser lens105and enters the light receiving element100A via the beam stopper106, the pinhole plate107, and the band pass filter108.

On the other hand, side scattered light and lateral fluorescence derived from the particles passing through the flow cell20are collected by the condenser lens109. The side scattered light enters the light receiving element100B via the dichroic mirrors110,111,112, the pinhole plate103, and the band pass filter114. The lateral fluorescence having a wavelength of 520 nm or more and 542 nm or less passes through the dichroic mirrors110and111, is reflected by the dichroic mirror112, and enters the light receiving element100C via the pinhole plate115and the band pass filter116. The lateral fluorescence having a wavelength of 570 nm or more and 620 nm or less passes through the dichroic mirror110, is reflected by the dichroic mirror111, and enters the light receiving element100D via the pinhole plate117and the bandpass filter118. The lateral fluorescence having a wavelength of 670 nm or more and 800 nm or less also is reflected by the dichroic mirror110, passes through the dichroic mirror119, and enters the light receiving element100E via the pinhole plate120and the bandpass filter121.

The light emitted from the light source124irradiates the flow cell20via the collimator lens125, the dichroic mirror103, and the condenser lens104. The lateral fluorescence of the light derived from the particles passing through the flow cell20is collected by the condenser lens109. The lateral fluorescent light of 662.5 nm or more and 687.5 nm or less is reflected by the dichroic mirror110, reflected by the dichroic mirror119, then enters the light receiving element100F via the pin hole plate122and the band pass filter123.

In the present embodiment, a laser diode with a wavelength of 488 nm is used for the light source101, and a laser diode with a wavelength of 642 nm is used for the light source124. A sheath flow cell is used for the flow cell20. A photodiode also is used for the light receiving element100A for receiving the forward scattered light, an avalanche photodiode (APD) is used for the light receiving element100B for receiving the side scattered light, and photomultiplier tubes (Photo Multiplier Tube, PMT) are used for light receiving elements100C to100F. In the present embodiment, the flow cytometer10includes six light receiving elements100A to100B, and the four light receiving elements100C to100F are for respectively detecting optical information of the four lights having different peak wavelengths derived from a stain bonded to particles in a sample, but the invention is not limited to this and it is also possible to provide three or more light receiving elements and at least two or more of the three or more light receiving elements may detect optical information of light originating from at least two stains having different peak wavelengths. For example, when four kinds of labeled antibody stains respectively binding to CD4, CD45, CD8, and CD3 are used in HIV examination, four fluorescent lights having four peak wavelengths derived from the respective labeled antibody stains are used as the measurement sample, and each fluorescence can be detected by the four light receiving elements100C to100F.

The number of light sources may be one, or two or more. The light source is selected according to the wavelength region of light derived from the stain bound to the particle. When the light sources are 2 or more, it is preferable that these light sources emit light having different peak wavelengths. Two light sources or more are preferable because it is possible to separate and detect fluorescence with high accuracy as compared with the case where there is only one light source. For example, when one light source is used in HIV testing, FITC is used as a labeled antibody dye for CD4 and PEcy5 is used as a labeled antibody dye for CD8; however, the peak wavelength of fluorescence from FITC and fluorescence from PEcy 5 are close to each other and the overlapping portions of the respective wavelength regions tend to be large. On the other hand, when using two light sources it is possible to separate and detect a plurality of fluorescences by shifting the light emission timing from each light source. It is also possible to reduce the overlapping portion of the respective wavelength regions of the plurality of fluorescences by using a dye suitable for the peak wavelength of light from each light source. For example, APC instead of PEcy 5 can be used as a labeled antibody dye for CD8. The number of photodiodes, dichroic mirrors, and bandpass filters can be varied according to the number of peak wavelengths of light derived from the particles. The types of photodiodes, dichroic mirrors, and bandpass filters can also be selected according to the peak wavelength of light derived from the particle, the wavelength region, and the intensity thereof.

As shown inFIG.3, the detection signals output from the light receiving elements100A to100F are amplified by the amplifying sections130A to130F, subjected to A/D conversion by the A/D conversion units131A to131F, and input to the signal processing unit63. Specifically, the amplifying units130A and130B connected to the light receiving element100A which is a photodiode and the light receiving element100B which is an APD are known amplifying circuits configured by operational amplifiers or the like, and the output voltages of the light receiving elements100A and100B to be input are adjusted by adjusting the amplification degree of each amplifier circuit. The output voltage of the PMT is adjusted by adjusting the voltage value applied to the light receiving elements100C to100F which are PMTs. Hereinafter, the adjustment of the detection sensitivity of the light receiving elements100A to100F refers to adjustment of the degree of amplification of the amplifying circuit in the light receiving elements100A and100B, and the adjustment of the voltage applied to the light receiving elements100C to100F in the light receiving circuits100C to100F. The detection signal output from the light receiving element is amplified by adjusting the amplification degree in the amplification circuit in the light receiving elements100A and100B, and the detection signal output by the light receiving elements100C to100F is adjusted by adjusting the voltage applied to the light receiving elements100C to100F. Note that amplification includes the case where the ratio of the output signal to the input signal is 1 or more and the case where the ratio is less than 1. Known amplification circuits also may be included in the amplifying units130C to130F connected to the light receiving elements100C to100F, and adjustment of the detection sensitivity of the light receiving elements100C to100F also may include adjusting the output voltages of the light receiving elements100C to100F via the amplification circuits.

The flow cytometer10is provided with the structure ofFIG.2that includes a light source124, the flow cell20, and the light receiving elements100A to100F, the amplifying sections130A to130F, and a measurement unit65that includes the A/D conversion units131A to131F, the signal processing unit63, and a temperature sensor22to be described later. The measurement unit65optically measures the particles in the particle-containing liquid passing through the flow cell20according to the measurement condition received by the communication unit64described later. Here, measurement includes detecting optical information of light derived from the particles by the light receiving elements100A to100F and storing the detection signals output by the light receiving elements100A to100F, and also includes processes performed by the signal processing unit63(to be described later) such as, for example, generating the particle count result and the like using the stored detection signals. The detection signals output from the light receiving elements100A to100F include signals output from the A/D conversion units131A to131F via the amplifiers130A to130F.

As shown inFIG.1A, the flow cytometer10includes a flow cytometer main body13and an information processing apparatus14connected to the flow cytometer main body13, and the structure ofFIG.2incorporating the light source124, flow cell20, and light receiving elements100A to100F of the measurement unit65, as well as the amplification units130A to130F, and the A/D conversion units131A to131F are arranged in the flow cytometer main body13. The signal processing unit63also is arranged in the information processing apparatus14. Note that, when the flow cytometer10does not include the information processing device14, the signal processing unit63may be disposed in the flow cytometer main body13. The flow cytometer10also has a control unit for controlling a pump, a motor and the like (not shown) in order to perform measurements by passing the particle-containing liquid through the flow cell20, although the signal processing unit63may also serve as this control unit, or this control unit may be separately arranged in the information processing apparatus14or the flow cytometer main body13.

Measurement Conditions

As described above, the flow cytometer10receives the measurement condition from the external server11in order to set the measurement condition according to the measurement item prior to the measurement.FIG.4shows an example of information included in the received measurement condition in the case where the light derived from the particle is fluorescent. The measurement condition includes basic information relating to measurement (hereinafter referred to as “basic measurement information”), information relating to adjustment of detection sensitivity for detecting optical information (hereinafter referred to as “information relating to adjustment of detection sensitivity”), information relating to correction of the detected optical information, information related to gating for setting an area of the particles to be selected based on the optical information (hereinafter referred to as “information related to gating”), and a calculation formula for use in temperature correction described later.

The basic measurement information includes basic information, measurement information, and a threshold value. The basic information includes identification information (referred to as “measurement condition ID” inFIG.4) for identifying the type of measurement condition and a measurement condition name. The measurement information includes the analytical capacity of the sample suctioned into the flow cytometer, the flow rate indicating the flow speed when the particles flow into the flow cytometer, and the dilution ratio of the sample suctioned into the flow cytometer. The threshold value is also called a detection level and is a set value of the lower limit of the optical information detected as particles. The threshold value is set for each light receiving element100A to100F with respect to light originating from the particle. For example, the threshold value can be set within a numerical value range of 0 to 1023 according to the intensity of light. If the threshold value is set at 50, light having a light intensity of 50 or more is detected as a particle.

Information relating to adjustment of the detection sensitivity includes at least one of a value indicating the amplification degree of the output voltage of the light receiving elements100A to100F, and a voltage value applied to the light receiving elements100A to100F. For example, an amplification value for adjusting the amplification degree of the amplification circuit connected to the light receiving elements100A and100B and a PMT voltage value for adjusting the voltage applied to the light receiving elements100C to100F. Note that the information may include only one of the amplification value and the PMT voltage value. When the amplification circuit is connected to the light receiving elements100C to100F, the information may include an amplification value for adjusting the amplification degree in the amplifying circuit.

The information related to the correction of the detected optical information includes information on the light wavelength distribution amount outside the detection target included in the optical information detected by the light receiving elements100A to100F. This is because when two or more lights with different peak wavelengths emitted by particles are detected in one measurement, the wavelength regions of the two or more lights may partially overlap one another. Therefore, the light which is not the detection target leaks into the light to be detected, and the specificity of detection of light may decrease. The distribution of the wavelength of the light and the amount of light are referred to as the light wavelength distribution amount, and the distribution of the wavelength of the leaked light and its amount are referred to as the non-detection target light wavelength distribution amount. Since the light receiving elements100C to100F can not distinguish the received overlapping portions of the wavelength ranges of two or more lights, so-called fluorescence correction is performed to remove the electrical signals originating from the non-detection target fluorescence, and capture only the optical information from the fluorescence to be detected from the electric signals of the light receiving elements100C to100F. Information relating to the light wavelength distribution amount outside the detection target included in the detected optical information is indicated as a fluorescence correction value inFIG.4, and is used for this fluorescence correction. The simplest fluorescence correction value is the light wavelength distribution amount of the fluorescence which is not to be detected that should be subtracted from the light wavelength distribution amount of the fluorescence to be detected. For example, when fluorescence having two different peak wavelengths is defined as fluorescence 1 and fluorescence 2, and if the light wavelength distribution does not overlap between fluorescence 1 and fluorescence 2 and there is no need for fluorescence correction, the fluorescence correction value of fluorescence 1 is 0.0. On the other hand, when fluorescence 1 and fluorescence 2 are simultaneously measured, the light distribution wavelengths overlap; when the overlapping light wavelength distribution amount is 27.5%, the fluorescence correction value can be set to 27.5 to subtract the fluorescence distribution amount derived from fluorescence 2 is calculated from the fluorescence distribution amount of fluorescence 1.

Information related to gating includes information related to a distribution setting on a distribution map of light derived from particles. In the flow cytometer, a distribution diagram of light originating from particles called a scattergram or a histogram, and is created for one measurement item or for each of two or more measurement items from the detected optical information. The scattergram represents the distribution of light originating from the particle on the two axes of X axis and Y axis for two measurement items. The histogram is represented by the intensity of light and the number of its particles for one measurement item. Gating refers to selection of a fixed distribution region corresponding to a measurement item in a distribution map for appropriate measurement according to the measurement item relative to the respective distribution map. More specifically, gating means setting the following information.

Information relating to the scattergram, information related to the histogram, and information related to gating is included in the distribution setting information on distribution of light derived from particles. The information relating to the scattergram is information for creating a scattergram, and includes the scattergram name which indicates the name of the scattergram to be created, the upper gate, X-axis channel (referred to as “X-axis channel”) indicating the photodiode receiving the light representing a first measurement item, X-axis channel name, Y-axis channel (referred to as Y-axis channel) indicating the photodiode receiving light representing a second measurement item, and Y-axis channel name. The information related to the histogram is information for creating a histogram, and includes a histogram name, an upper gate, an X axis channel indicating the photodiode receiving light representing the measurement item, and X-axis channel name. The upper gate indicates the gate of the previously created scattergram when creating respectively corresponding scattergrams using two or more gates. The information related to the gate is for determining the area of each particle selected from the scattergram and the histogram, and includes a gate name which is the name of the selected gate, position information indicating the position of the gate, a color given on the display unit for the received light wavelength or wavelength range, the measurement item name, the upper limit value of the intensity of the received light, the lower limit value of the intensity of the received light, and the result value type when displaying the analysis result. The result value types are various statistical processing values of the result, for example, the total number of particles, the average value, the variation coefficient, the ratio to the whole, the mode, and the like.

The number of scattergrams and histograms created differs depending on measurement items. Therefore, there are cases where a plurality of scattergram related information, histogram related information, and gate related information are included in accordance with the number of scattergrams and histograms to be created.

Information Processing System of Flow Cytometer

Hereinafter, the instance where the light derived from particles is fluorescent will be specifically described as an example.

FIG.3shows the configuration of the information processing system of the flow cytometer10, which includes an input unit60, a condition input unit61, a display unit62, a signal processing unit63, and a communication unit64. The signal processing unit63acquires the detection signals output from the light receiving elements100A to100F via the amplifiers130A to130F and the A/D converters131A to131F. A temperature sensor22is provided to detect the temperature of the particle-containing liquid and output a temperature detection signal converted into an electric signal; the signal processing unit63acquires the temperature detection signal from the temperature sensor22through the temperature detection circuit132and the A/D conversion unit133.

The input unit60is configured by a barcode reader and accepts input of information for specifying measurement items by reading a barcode attached to a container containing a reagent to be mixed with particles. Note that the input unit60may be an RFID reader that reads information for specifying a measurement item from a tag attached to a container. The input unit60also may be configured by at least one of a keyboard and a mouse, and the user may manually input information for specifying measurement items or select from among a plurality of options prepared in advance.

For example, the condition input unit61may be configured with at least one of a keyboard, a mouse, and a touch panel to accept input of measurement conditions when a user inputs measurement conditions.

The display unit62is configured by, for example, a monitor, and displays the screens90to96shown inFIGS.6to12and the analysis result.

The input unit60, the condition input unit61, and the display unit62are disposed in the information processing apparatus14connected to the flow cytometer main body13, but also may be arranged in the flow cytometer main body13.

The communication unit64is configured by a communication device for communicating with the external server11via the network12according to a predetermined communication standard. Information for specifying measurement items is transmitted to the external server11via the network12, and measurement conditions and the like corresponding to information for specifying measurement items are received from the external server11via the network12.

The signal processing unit63includes a memory82used as a work area for data processing, a storage unit83for recording programs and processing data, a CPU (Central Processing Unit)81for performing data processing described later, a bus84for relaying data between units, and interface units (I/F units inFIGS.3)85and86which perform data input/output between the respective units60,61,62, and64connected to the signal processing unit63, or inputs detection signals output from the light receiving devices100A to100F via the amplifiers130A,130F and A/D conversion units131A to131F, or inputs a temperature detection signal from the temperature sensor22via the temperature detection circuit132and the A/D conversion unit133.

In the following description, unless otherwise specified, the processing performed by the signal processing unit63actually means the processing performed by the CPU81of the signal processing unit63. The CPU81temporarily stores necessary data (such as intermediate data being processed) using the memory82as a work area, and records data to be stored longterm in the storage unit83.

By executing a program stored in the storage unit83or the memory82, the signal processing unit63performs data processing to realize the functions of a measurement item acquisition unit70, a measurement condition selection unit71, a measurement condition storage unit72, a first measurement condition setting unit73, a temperature correction unit74, a data processing unit75, a second measurement condition setting unit76, and a calculation unit77, and controls the operation of each unit connected to the signal processing unit63.

Operation of Information Processing System of Flow Cytometer

FIG.13is a flowchart illustrating the operation of the signal processing unit63and the external server11of the flow cytometer10of the invention. The measurement item acquisition unit70performs the process ST10, the measurement condition selection unit71performs process ST11to ST16, the measurement condition storage unit72performs processes ST17and ST18, the first measurement condition setting unit73and temperature correction unit74perform the process of ST19, data processing unit75performs the process ST20, and the process ST21is performed by the calculation unit77. The processes from ST50to ST54are performed in the external server11. Note that the information that the signal processing unit63exchanges with the external server11is performed via the communication unit64.

In ST10, the signal processing unit63acquires information for specifying the measurement item. For example, using the barcode reader as the input unit60, information for specifying the measurement item is read from the barcode attached to the reagent container. The information for specifying the measurement item is, for example, the identification information of the reagent mixed with the particle. In ST11, the signal processing unit63transmits information for specifying the measurement item to the external server11. Note that the barcode attached to the reagent container also may include manufacturer identification information for identifying the manufacturer of the reagent. In that case, the measurement condition is received from the external server11only when manufacturer identification information indicating a specific manufacturer is acquired from the bar code of the reagent container, and a notice indicating a measurement condition is not received is displayed on the display unit62when the manufacturer identification information indicating a specific manufacturer is not obtained.

In ST50, the external server11receives information for specifying the measurement item. In ST51, the external server11searches whether a measurement condition corresponding to information (identification information of the reagent) for specifying this measurement item exists in the database and, in ST52, the search result is transmitted to the signal processing unit63. The search result is either information on a list (hereinafter referred to as “measurement condition candidate list”) showing one or more measurement condition candidates corresponding to the information (identification information of the reagent) for specifying the measurement item, or information indicating that there is no corresponding measurement condition. The measurement condition candidate list includes the measurement condition name, measurement condition description and the like corresponding to the measurement items that can be measured using the reagent in the reagent container from which the identification information of the reagent was read in ST10. For example, when the identification information of the reagent containing the labeled antibody against CD4, the labeled antibody against CD45 or the like is read in ST10, the measurement conditions corresponding to the HIV test, the hematopoietic stem cell test and the like that can be carried out using these labeled antibodies are included in the list.

In ST12, the signal processing unit63receives the search result from the external server11via the network12. In ST13, the signal processing unit63determines whether information on the measurement condition candidate list has been received, or information indicating that there is no measurement condition has been received. Upon receiving the measurement condition candidate list corresponding to the information for specifying the measurement item, the signal processing unit63displays screen90that shows the measurement condition candidate list shown inFIG.6on the display unit62in ST14. When there is no measurement condition corresponding to the information for specifying the measurement item in the database, the process proceeds to ST31shown inFIG.14. In ST15, upon receiving the measurement condition candidate selected from the measurement condition candidate list by the user, the signal processing unit63transmits the accepted measurement condition candidate to the external server11via the network12in ST16.

In ST53, the external server11receives the measurement condition candidate selected by the user, and in ST54transmits the measurement condition corresponding to the selected measurement condition candidate. In ST17, the signal processing unit63receives the measurement condition corresponding to the selected measurement condition candidate from the external server11via the network12, and stores it in the storage unit83. In the storage unit83, the name indicating the measurement item and the measurement condition are associated and stored.

When there is only one measurement condition corresponding to the information for specifying the measurement item in the external server11, the signal processing unit63also may receive and store the measurement condition without receiving a measurement condition candidate list from the external server11. That is, ST52and ST53are skipped, and ST12to ST16are skipped.

In ST18, the signal processing unit63accumulates the measurement conditions received from the external server11. That is, the received measurement condition is added to the measurement condition list in which the measurement conditions received in the past are stored, and this list is stored. In ST19, the signal processing unit63sets measurement conditions. The setting of the measurement conditions entails adjusting the amplification degree of the output voltage of the light receiving element or the voltage applied to the light receiving element by using the information relating to the adjustment of the detection sensitivity, and determining the amount of light wavelength distribution outside the detection target included in the detected optical information (referred to as “fluorescence correction setting”) using information relating to the correction of the detected optical information, and determining the region of each particle to be selected from the distribution chart by using the information related to the gating.

Detection signals output from the light receiving elements100A to100F are input to the signal processing section63via the amplifying sections130A to130F and the A/D converting sections131A to131F. In ST20, the signal processing unit63performs correction of this detection signal, that is, so-called fluorescence correction, by using the light wavelength distribution amount outside the detection target included in the detected optical information. The signal processing unit63also reads out a calculation formula used for temperature correction from the measurement conditions stored in the storage unit83, and uses the temperature detection signal output from the temperature sensor22and the calculation formula to correct the detection signals output from the light receiving elements100A to100F, which will be described later in detail. In ST21, the signal processing unit63performs analyses including the measurement of the number of particles using the fluorescence correction and the temperature-corrected detection signal, and outputs the analysis result. The output of the analysis result is, for example, displaying the analysis result on the display unit62together with the distribution chart.

Note that the signal processing unit63also may store the input detection signal in the storage unit83without displaying the analysis and the analysis result of ST21, and the communication unit64may transmit the detection signal stored in the storage unit83to external server11or another external server other than the external server11, such that the analysis performed by the external server11or another external server other than the external server11can be displayed as analysis results.

FIG.14is a flow chart describing the sequence of the operation of accepting input of a measurement condition by a user when the communication unit64does not receive a measurement condition from the external server11because there is no measurement condition corresponding to the information for specifying a measurement item in the external server11. The processes from ST31to ST40are performed by the second measurement condition setting unit76. In the processes from ST31to ST40, the screens91to95for the user to perform the input operation of the measurement condition from the condition input unit61are sequentially displayed on the display unit62in the wizard format, and the measurement conditions input by the user are set.

In ST31, the signal processing unit63displays the new creation screen91shown inFIG.7on the display unit62. In ST32, the signal processing unit63accepts the name of the measurement condition and comment and the like input by the user in the input box on the screen91.

When the user presses a “next” button91aon the screen91and the signal processing unit63accepts a screen switching instruction, the signal processing unit63displays on the display unit62a screen (hereinafter referred to as “sensitivity input screen”)92for inputting information relating to adjustment of the detection sensitivity in order to detect the optical information shown inFIG.8in ST33. The sensitivity input screen92is information related to adjustment of the detection sensitivity, and is a screen for allowing the user to input information related to the size of the particles in the particle-containing liquid, for example, the size of the particles such as the diameter of the particle, the name of the particle, the name of the measurement item and the like. Also, the particle size may be input in three stages of “large”, “medium” and “small”.

In ST34, the signal processing unit63receives information on the size of the particles in the particle-containing liquid which the user inputs in the input box on the screen92. The storage unit83stores a value indicating the amplification degree of the output voltage of the light receiving elements100A and100B corresponding to the information on the size of the particles in the particle containing liquid and stores the voltage value applied to the light receiving elements100C to100F, and the signal processing unit63reads the value indicating the amplification degree corresponding to the particle size and the voltage value from the storage unit83as information relating to adjustment of the detection sensitivity.

When the signal processing unit63accepts a screen switching instruction by the user pressing the “Next” button92aon the screen92, in ST35the signal processing unit63displays the detected optical information shown inFIG.9(hereinafter referred to as “fluorescence correction value input screen”)93on the display unit62. In ST36, the signal processing unit63receives information (also referred to as “fluorescence correction value”) relating to the light wavelength distribution amount other than the detection target included in the detected optical information. The fluorescence correction value is entered by the user in an input box on this screen93. When the signal processing unit63accepts a screen switching instruction by the user pressing the “Next” button93aon the screen93, in ST37, the signal processing unit63displays on the display unit62a screen (hereinafter referred to as “gating input screen”)94for setting an area of selected particles based on the optical information, as shown inFIG.10. In ST38, the signal processing unit63receives information related to gating that the user input in the input box on the screen94.

When the user presses the “completion” button94aon the screen94and the signal processing unit63accepts the input completion instruction, then in ST39the signal processing unit63stores the value indicating degree of amplification of the output voltage of the light receiving elements100A and100B, voltage value applied to the light receiving elements100C to100F, fluorescence correction value, and information related to gating from the user as measurement conditions. In ST40, the signal processing unit63accumulates measurement conditions input by the user. That is, the measurement condition input by the user is added to the measurement condition list in which the measurement conditions received in the past are accumulated, and this list is stored. Note that a list of measurement conditions in which both measurement conditions input by the user and measurement conditions received from the external server11are both stored also may be stored, and a measurement condition list of accumulated measurement conditions input by the user and a measurement condition list of accumulated measurement conditions received from the external server11also may be stored.

Returning to ST19inFIG.13, adjustments of the detection sensitivity, fluorescence correction, and gating are set using the stored measurement conditions.

When inputting each information on the screens92,93,94, the user preliminarily measures a control sample consisting of artificially created components such as a single-dye control sample stained only with FITC, for example, one or more times, then, based on the measurement result, a value indicating an appropriate amplification degree, a voltage value, a fluorescence correction value, and information related to gating may be determined. Upon receiving a measurement instruction of the control sample from the user while each screen92,93,94is displayed, the signal processing unit63causes the measurement instruction screen95shown inFIG.11to be displayed on the screens92,93, and94. When the measurement sample is selected according to the instruction of the screen95, the signal processing unit63confirms that the control sample is set at the predetermined position, and starts the preliminary measurement according to the measurement instruction. When the user presses the “remeasure” buttons92c,93c,94con the screens92,93,94, the signal processing unit63performs the preliminary measurement again. On screen93, when the user presses the “redraw” button93bon the screen93, the signal processing unit63redisplays the distribution diagram of the scattergram and the like on the screen93.

The order of displaying the screens92,93, and94is not limited, and may be in any order.

When it is desired to reuse the measurement condition used in the past after the completion of the measurement, the signal processing unit63calls up the accumulated measurement condition list according to the user's request, and displays a screen96that displays the measurement condition list shown inFIG.12, and displays this list on the display unit62. Upon receiving the measurement condition selected from the measurement condition list by the user, the signal processing unit63sets this measurement condition in the same manner as ST19.

Temperature Correction by Temperature Sensor

Regarding the temperature correction of the detection signals of the light receiving elements100A to100F, a method of detecting the temperature of the particle containing liquid by the temperature sensor22will be described.

The flow cell20shown inFIG.15includes a rectification part23, an acceleration part24, and an orifice part25. The rectification part23is cylindrical and has a through hole. The acceleration part24is conical, and the diameter of the hole communicating with the hole of the rectification part23gradually decreases toward the orifice part25. The orifice part25of the flow cell20is a transparent rectangular tube having a square cross section, and the particle-containing liquid passing through the hole is irradiated with the light L from the light sources101and124.

The flow cell20is fixed to the fixture26, and the nozzle21is inserted into the flow cell20and attached to the fixture26. A sensor setting member27having a flow path27ais connected to an outlet of the orifice part25of the flow cell20. The sensor setting member27is provided with a temperature sensor22and a nipple28. The particle-containing liquid introduced into the flow path27afrom the orifice part25contacts the temperature sensor22and is discharged from the nipple28. In this case, a thermistor (PB3M-35-TI type manufactured by Shibaura Electronics Co., Ltd.) is used as the temperature sensor22.

The sheath liquid flowing in from the inflow port26aof the fixture26is rectified by the rectification part23, and the flow is accelerated by the acceleration part24. When the sample flowing into the nozzle21from the direction of the arrow B is ejected from the tip of the nozzle21toward the orifice part25, the sample is enveloped by the accelerated sheath liquid, and passes through the orifice part25as a particle-containing liquid. Light L is irradiated on the particle-containing liquid, and light derived from the particles in the particle-containing liquid is detected by light receiving elements100A to100F shown inFIG.2.

In the sensor setting member27, the liquid temperature of the particle-containing liquid that has passed through the orifice section25is detected by the temperature sensor22, and the liquid is discharged from the nipple28in the direction of arrow C.

Note that the above-described mounting position of the temperature sensor22is an example, and the temperature sensor22also may be provided, for example, at the entrance or the vicinity of the flow cell20as long as it is attached to a certain position, or it may be provided outside the flow cell10insofar as the temperature sensor22is attached at a position where the temperature of the particles to be detected is substantially the same as the temperature of the particle-containing liquid passing through the orifice part25.

The temperature correction of the detection signals of the light receiving elements100A to100F will be described next. Temperature correction is performed by the temperature correction unit74of the signal processing unit63, for example, by using the temperature detection signal converted into the electric signal output from the temperature sensor22, and amplifying the detection signals of the light receiving elements100A to100F in each amplification unit130A to130F connected to each light receiving element100A to100F. The signal processing unit63acquires the temperature detected by the temperature sensor22via the thermistor circuit132and A/D conversion unit133.

Detection signals of the light receiving elements100A to100F are input to the amplifying sections130A to130F. A 0.5 V constant voltage is input to the amplification unit as the low level reference voltage RL, and a control voltage is input from the signal processing unit63as the high level/reference voltage RH.

A correction curve (actually measured values) of the detection signal of the light receiving elements100A to100F relative to the temperature detected by the temperature sensor22shows the relationship of the control voltage to the temperature (° C.). For example, when the measurement item is reticulocyte, it is expressed as calculation formula (A). y=4.4838 (x−23)2−64.815 (x−23)+3031 . . . (A), where x is the temperature (° C.) detected by the temperature sensor22, and y is the control voltage (digital value). The calculation formula (A) is received from the external server11as a measurement condition for each measurement item.

The ratio of the output to the input of the A/D converter, that is, the amplification factor, is determined by the difference (RH−RL) between RH and RL. Since RL is constant, the degree of amplification is controlled by RH, that is, the control voltage determined by the calculation formula (A), and the detection signals of the light receiving elements100A to100F are temperature-corrected by the detected temperature of the temperature sensor22.

Although the amplification degree of the amplification units130A to130F is controlled in the above embodiment, the temperature correction method is not limited to this method, inasmuch as, for example, the respectively voltage applied to the light receiving elements100A to100F also may be adjusted based on the temperature detection signal of the temperature sensor22. The analysis result obtained by the calculation unit77also may be corrected.

Note that the temperature sensor22, the temperature detection circuit132, the A/D conversion unit133, and the temperature correction unit74of the signal processing unit63also may be omitted so that the temperature correction is not performed.

According to the invention, an appropriate measurement condition is selected from one or a plurality of measurement conditions according to measurement items. For example, a measurement condition is selected from one or a plurality of measurement conditions received from the external server11. Therefore, it is not necessary for the user to input complicated measurement conditions himself. Therefore, measurement conditions can be easily set even by a non-specialist technician who has not received specialized training. In the case where there is no measurement condition corresponding to the information for specifying the measurement item in the external server and the measurement condition can not be received, the user can set the measurement condition from the condition input unit61.

The flow cytometer10of another embodiment includes a temperature sensor for detecting the temperature inside the flow cytometer10and outputting a temperature detection signal converted into an electric signal, and a temperature adjustment device for heating or cooling the interior of the flow cytometer10provided at an arbitrary position within the flow cytometer10. The temperature adjustment device includes, for example, at least one of a heater and a fan. The signal processing unit63does not include the temperature correction unit74. The temperature adjustment device is connected to the signal processing unit63, and the temperature adjustment device performs controls so that the temperature in the flow cytometer10becomes a target temperature using the temperature detection signal output from the temperature sensor22. The target temperature may be included in the measurement condition received from the external server11by the signal processing unit63, or may be stored in advance in the signal processing unit63. Since the temperature inside the flow cytometer10is kept constant, the signal processing unit63does not need to perform temperature compensation on the detection signal output from the light receiving elements100A to100F. Once the measurement condition corresponding to the information for specifying the measurement item is received and the measurement is started, there is no need for the user to change the measurement condition according to the temperature. Since other configurations are the same as those of the flow cytometer10shown inFIGS.1to15, description thereof is omitted.

Although one embodiment of the present invention has been described above, the present invention is not limited to the above-described embodiment, and various modifications are possible insofar as the modifications do not deviate from the spirit of the invention

For example, although one or a plurality of measurement conditions corresponding to the identification information of the reagent are received from the external server11, and the measurement condition is selected from among them in the embodiment, one or a plurality of measurement conditions corresponding to the identification information of the reagent also may be previously stored in the storage unit83of the flow cytometer10and the measurement condition may be selected from one or more measurement conditions read out from the storage unit83; and measurement conditions also may be stored in advance in a storage unit of an external computer installed in the facility of the user, for example, a host computer that manages the examination room, and the flow cytometer10may receive the measurement condition from the host computer.