JP2019052982A - Cell mass measuring apparatus, analyzer and cell mass measuring method - Google Patents

Abstract

To provide a cell mass measuring apparatus capable of measuring the amount of cells of a small amount sample, an analyzer including the same and a cell mass measuring method.SOLUTION: A sample 10 containing the bacteria to be measured is held on the sample plate 220. A light projector 230 irradiates the sample 10 held on the sample plate 220 with a light beam. An intensity acquisition part D acquires the intensity of the scattered light from sample. A transformation information acquisition part E acquires a piece of transformation information which represents the correspondence between the intensity of scattered light and the amount of cells. A cell amount determination part F determines the amount of the cells in the sample based on the intensity of the scattered light acquired by the intensity acquisition unit D and the conversion information acquired by the conversion information acquisition unit E.SELECTED DRAWING: Figure 2

Description

  The present invention relates to a bacterial cell amount measuring apparatus for measuring a bacterial cell amount, an analysis apparatus including the same, and a bacterial cell amount measuring method.

  When analyzing bacteria with an analyzer, the number of signal integrations is adjusted based on information on the amount of cells so that an appropriate signal-to-intensity ratio of the analysis results can be obtained. The amount of bacterial cells refers to the concentration of bacteria in the sample or the number of bacteria in the sample. Moreover, when performing quantitative evaluation of analysis data, the information regarding the amount of microbial cells is used. Thus, in the analysis of bacteria, information on the amount of cells is required. Non-Patent Document 1 introduces various methods such as dry cell weight method and turbidity method as methods for measuring cell concentration.

Masanori Konishi and Junichi Horiuchi, “Capturing Cell Growth—From Measurement Methods to Specific Velocity Calculations”, Journal of Biotechnology, 2015, Vol. 93, No. 3, p.149-152

  In the method introduced in Non-Patent Document 1, in order to measure the cell concentration, a relatively large amount (for example, about 0.5 to 1 mL) of a bacterial specimen amount is required. On the other hand, the amount of bacterial specimen required for analysis by the analyzer is very small (for example, about 1 μL). Therefore, conventionally, a small amount of sample containing bacteria to be measured has been introduced into the analyzer by dispensing a relatively large amount of sample in which the amount of bacterial cells has been measured. However, since the remaining sample is discarded without being used for analysis, the amount of waste increases.

  The objective of this invention is providing the microbial cell amount measuring apparatus which can measure the microbial cell amount of a trace amount sample, an analyzer provided with the same, and a microbial cell amount measuring method.

  (1) A bacterial cell amount measuring device according to a first invention is a bacterial cell amount measuring device for measuring a bacterial cell amount, a sample plate for holding a sample containing bacteria to be measured, and a sample plate A first light projecting unit that irradiates the held sample with a light beam, an intensity obtaining unit that obtains the intensity of scattered light from the sample, and conversion information that indicates the correspondence between the intensity of the scattered light and the amount of bacterial cells. A conversion information acquisition unit; and a bacterial cell amount specifying unit that specifies the bacterial cell amount in the sample based on the intensity of the scattered light acquired by the intensity acquisition unit and the conversion information acquired by the conversion information acquisition unit.

  In this bacterial cell amount measuring apparatus, light is scattered on the surface of the bacteria in the sample by irradiating the sample containing the bacteria to be measured held on the sample plate with a light beam. The intensity of scattered light from the sample is acquired, and conversion information indicating the correspondence between the intensity of scattered light and the amount of bacterial cells is acquired. Based on the acquired scattered light intensity and conversion information, the amount of bacterial cells in the sample is specified. According to this configuration, the intensity of scattered light from the sample can be acquired even when the sample is a very small amount. Thereby, the microbial cell quantity of a trace amount sample can be measured.

  (2) The device for measuring the amount of bacterial cells includes the intensity of the scattered light acquired by the intensity acquisition unit and the bacteria in the reference sample in a state where a reference sample containing bacteria having a known amount of bacterial cells is held on the sample plate. You may further provide the conversion information production | generation part which produces | generates conversion information based on body weight. In this case, conversion information can be easily generated. In addition, using the generated conversion information, it is possible to specify the amount of bacterial cells in the sample containing bacteria that are the same type as the bacteria in the reference sample and whose bacterial mass is unknown.

  (3) At least one of the number of the first light projecting units, the irradiation position of the emitted light beam by the first light projecting unit, the emission direction of the emitted light beam, the diameter of the emitted light beam, and the wavelength of the emitted light beam is configured to be changeable. Also good. In this case, depending on the type, form, and characteristics of the bacteria in the sample, the number of the first light projecting units, the irradiation position of the emitted light beam by the first light projecting unit, the emission direction of the emitted light beam, the diameter of the emitted light beam, or The wavelength of the emitted light beam can be changed.

  (4) The sample plate may include a well for holding the sample, and the diameter of the light beam emitted by the first light projecting unit may be set so as to include the well. In this case, the emitted light beam can be reliably and easily irradiated to the bacteria in the sample.

  (5) The bacterial cell amount measuring device further includes a light receiving unit that receives scattered light from the sample and generates a light receiving signal indicating the amount of received light, and the intensity acquisition unit is based on the light receiving signal generated by the light receiving unit. The intensity of scattered light may be acquired. In this case, the intensity acquisition unit can easily acquire the intensity of the scattered light.

  (6) At least one of the number of light receiving parts, the light receiving position by the light receiving parts, the light receiving direction, and the wavelength dependency may be configured to be changeable. In this case, the number of light receiving parts, the light receiving position by the light receiving part, the light receiving direction, or wavelength dependence can be changed according to the type, form, and characteristics of the bacteria in the sample.

  (7) The first light projecting unit and the light receiving unit may be arranged such that the optical axis of the first light projecting unit and the optical axis of the light receiving unit are asymmetric with respect to the normal to the surface of the sample plate. In this case, the light receiving unit can efficiently receive the scattered light from the sample without receiving the light regularly reflected by the sample plate.

  (8) The conversion information indicates a correspondence relationship between the intensity of scattered light and the amount of bacterial cells under a predetermined measurement condition, and the bacterial cell amount measurement device is based on the measurement conditions in the first light projecting unit or the light receiving unit. The conversion information correction unit for correcting the conversion information acquired by the conversion information acquisition unit is further provided, and the bacterial cell amount specifying unit converts the intensity of the scattered light acquired by the intensity acquisition unit and the conversion information corrected by the conversion information correction unit. The amount of bacterial cells in the sample may be specified based on the information. In this case, the amount of bacterial cells in the sample can be specified based on the conversion information even under measurement conditions different from the predetermined measurement conditions.

  (9) The analysis device according to the second invention includes a bacterial cell amount measurement device according to the first invention, and an analysis unit that analyzes a bacterial component in the sample held on the sample plate of the bacterial cell amount measurement device; Is provided. In this case, the measurement of the amount of bacterial cells in the sample and the analysis of the components of the bacteria in the sample can be performed using a common sample plate. Therefore, after the amount of microbial cells in the sample is measured by the microbial cell amount measuring device, it is not necessary to transfer the sample from the sample plate to another sample stage for component analysis. Thereby, the convenience of the analyzer is improved. In addition, since the measurement of the amount of bacterial cells in the sample and the analysis of the bacterial components in the sample are performed in the same state, it is possible to accurately grasp the relationship between the amount of bacterial cells in the sample and the analysis result of the sample. it can.

  (10) The analyzer includes a cell amount information acquisition unit that acquires cell amount information indicating the amount of cells in the sample specified by the cell amount specifying unit of the cell amount measurement device, and cell amount information acquisition An analysis condition setting unit that sets analysis conditions of the analysis unit based on the bacterial cell amount information acquired by the unit may be further provided. In this case, since analysis conditions are set based on the amount of bacterial cells in the sample measured by the bacterial cell amount measuring device, it is possible to appropriately analyze the components of the bacteria regardless of the amount of bacterial cells in the sample. .

  (11) The analysis unit outputs a signal indicating the analysis result, and the analysis device generates analysis data based on the signal output from the analysis unit and integrates analysis data generated by a plurality of analyzes A data processing unit may be further provided, and the analysis condition setting unit may set the number of integrations in the analysis data processing unit as the analysis condition. In this case, analysis data is integrated an appropriate number of times according to the amount of bacterial cells in the sample. Thereby, the component of bacteria can be analyzed appropriately regardless of the amount of bacterial cells in the sample.

  (12) The analysis unit includes a second light projecting unit that irradiates the sample held on the sample plate with light, and the first light projecting unit and the second light projecting unit of the bacterial cell amount measuring apparatus are common. The light projecting unit may be configured. In this case, it is not necessary to provide the first light projecting unit and the second light projecting unit separately. Thereby, the cost of an analyzer can be reduced. Moreover, the space for installing the light projection part in an analysis part can be made compact.

  (13) A method for measuring the amount of bacterial cells according to a third aspect of the present invention is a method for measuring the amount of bacterial cells, which measures the amount of bacterial cells, and the luminous flux is applied to a sample containing bacteria to be measured held on the sample plate by the light projecting unit. , A step of acquiring the intensity of scattered light from the sample, a step of acquiring conversion information indicating a correspondence relationship between the intensity of scattered light and the amount of bacterial cells, and the acquired intensity of scattered light and Identifying the amount of bacterial cells in the sample based on the converted information.

  According to this method for measuring the amount of bacterial cells, the intensity of scattered light from a sample can be acquired even when the sample is in a very small amount. Thereby, the microbial cell quantity of a trace amount sample can be measured.

  According to the present invention, the amount of bacterial cells in a small amount of sample can be measured.

It is a figure which shows the structure of the analyzer which concerns on one embodiment of this invention. It is a figure which shows the structure of the microbial cell amount measuring apparatus containing the microbial cell amount measuring part of FIG. It is a figure which shows an example of preferable measurement conditions. It is a figure which shows the detailed structure of the analyzer of FIG. It is a flowchart which shows the algorithm of the measurement process performed by a microbial cell amount measurement program. It is a flowchart which shows the algorithm of the mass spectrometry process performed by a mass spectrometry program.

  Hereinafter, a microbial cell amount measuring apparatus, an analysis apparatus including the same, and a microbial cell amount measuring method according to an embodiment of the present invention will be described in detail with reference to the drawings.

(1) Configuration of Analyzer FIG. 1 is a diagram showing a configuration of an analyzer according to an embodiment of the present invention. As shown in FIG. 1, the analysis device 1 includes a control device 100, a microbial cell amount measurement unit 200, and an analysis unit 300. In FIG. 1, the hardware configuration of the analyzer 1 is mainly shown.

  The control device 100 includes a CPU (central processing unit) 110, a RAM (random access memory) 120, a ROM (read only memory) 130, a storage device 140, an operation unit 150, a display unit 160, and an input / output I / F (interface). 170. The CPU 110, RAM 120, ROM 130, storage device 140, operation unit 150, display unit 160, and input / output I / F 170 are connected to the bus 101.

  The RAM 120 is used as a work area for the CPU 110. The ROM 130 stores a system program. The storage device 140 includes a storage medium such as a hard disk or a semiconductor memory, and stores a bacterial mass measurement program, a mass analysis program, conversion information, and the like. The conversion information will be described later.

  When the CPU 110 executes the bacterial cell amount measurement program stored in the storage device 140 on the RAM 120, a bacterial cell amount measurement process (hereinafter abbreviated as a measurement process) described later is performed. Further, when the CPU 110 executes the mass analysis program stored in the storage device 140 on the RAM 120, mass analysis processing described later is performed.

  The operation unit 150 is an input device such as a keyboard, a mouse, or a touch panel. The display unit 160 is a display device such as a liquid crystal display device. The user can give various instructions to the control device 100 using the operation unit 150. The display unit 160 can display the measurement result of the bacterial cell amount or the analysis result of the sample. The input / output I / F 170 is connected to the bacterial cell amount measurement unit 200 and the analysis unit 300. In the present embodiment, the bacterial cell amount measurement unit 200 is provided in the analysis unit 300. The bacterial cell amount measurement unit 200 may be provided independently of the analysis unit 300.

(2) Bacterial Cell Amount Measuring Device FIG. 2 is a diagram showing a configuration of a bacterial cell amount measuring device including the microbial cell amount measuring unit 200 of FIG. As shown in FIG. 2, the bacterial cell amount measuring device 2 includes a movable stage 210, a sample plate (sample stage) 220, a light projecting unit 230, a light receiving unit 240, and a measurement control unit 250. The movable stage 210, the sample plate 220, the light projecting unit 230, and the light receiving unit 240 constitute the bacterial cell amount measuring unit 200. The bacterial cell amount measuring unit 200 may include a plurality of light projecting units 230 or a plurality of light receiving units 240.

  The movable stage 210 includes an actuator and is movable in three directions orthogonal to each other. A sample plate 220 is placed on the movable stage 210. The sample plate 220 is a well plate in which a plurality of wells (see FIG. 3 to be described later) is formed. As the MALDI (matrix-assisted laser desorption / ionization) plate, the measurement of the amount of cells by the analyzer 1 in FIG. Commonly used for analysis. A small amount of the sample 10 containing bacteria to be measured is dropped into a desired well of the sample plate 220.

  The light projecting unit 230 includes a plurality of light sources and actuators. The plurality of light sources are laser light sources that emit light beams having different wavelengths, for example. Hereinafter, the light beam emitted by the light projecting unit 230 is referred to as an emitted light beam. The light projecting unit 230 is configured to be able to change the irradiation position of the outgoing light beam, the outgoing direction of the outgoing light beam, the diameter of the outgoing light beam, and the wavelength of the outgoing light beam. When the light projecting unit 230 irradiates the sample 10 with light, the light irradiated on the sample 10 is reflected by the irregularities on the surface of the bacteria and scattered radially around the sample 10.

  The light receiving unit 240 includes a plurality of light receiving elements and actuators. The plurality of light receiving elements are photomultiplier tubes or photodiodes having wavelength dependence with different light receiving sensitivities. The light receiving unit 240 is configured to be able to change the light receiving position, the light receiving direction, and the wavelength dependency. The light receiving unit 240 receives scattered light from the sample 10 and generates an electrical signal (hereinafter referred to as a light receiving signal) corresponding to the amount of light received.

  The measurement control unit 250 includes a stage control unit A, a light projection control unit B, a light reception control unit C, an intensity acquisition unit D, a conversion information acquisition unit E, a bacterial cell amount identification unit F, a measurement result output unit G, and a conversion information generation unit. H and conversion information correction unit I are included. The functions of the components (A to I) of the measurement control unit 250 are realized by the CPU 110 in FIG. 1 executing the bacterial cell amount measurement program stored in the storage device 140. Part or all of the components (A to I) of the measurement control unit 250 may be realized by hardware such as an electronic circuit.

  As described above, conversion information is stored in the storage device 140 in advance. The conversion information is information indicating the correspondence between the intensity of scattered light and the amount of bacterial cells under specific measurement conditions, and varies depending on the measurement conditions and the type of bacteria. The following measurement process is performed under the same measurement conditions as the measurement conditions in the conversion information.

  The user can designate the position of the movable stage 210 by operating the operation unit 150. The stage control unit A controls the movable stage 210 so that the movable stage 210 moves to a designated position based on the operation of the operation unit 150 by the user.

  By operating the operation unit 150, the user can specify the irradiation position of the emitted light beam by the light projecting unit 230, the emission direction of the emitted light beam, the diameter of the emitted light beam, and the wavelength of the emitted light beam. Based on the operation of the operation unit 150 by the user, the light projection control unit B controls the irradiation position of the emitted light beam by the light projecting unit 230, the emission direction of the emitted light beam, the diameter of the emitted light beam, and the wavelength of the emitted light beam.

  The user can designate the light receiving position, the light receiving direction, and the wavelength dependency by the light receiving unit 240 by operating the operation unit 150. The light reception control unit C controls the light reception position, the light reception direction, and the wavelength dependency by the light reception unit 240 based on the operation of the operation unit 150 by the user.

  The intensity acquisition unit D acquires the intensity of scattered light based on the light reception signal generated by the light receiving unit 240. The conversion information acquisition unit E acquires conversion information about the same bacteria as the sample 10 from the storage device 140. When an external storage medium is connected to the control device 100 in FIG. 1, the conversion information acquisition unit E may acquire conversion information stored in the external storage medium. When the control device 100 is connected to a network, the conversion information acquisition unit E may acquire conversion information stored in another storage device via the network.

  The bacterial cell amount identification unit F identifies the bacterial cell amount in the sample 10 based on the intensity of the scattered light acquired by the intensity acquisition unit D and the conversion information acquired by the conversion information acquisition unit E. The measurement result output unit G outputs the cell amount information indicating the cell amount specified by the cell amount specifying unit F to the storage device 140 and the display unit 160. Thereby, the bacterial cell amount information is stored in the storage device 140. Further, based on the bacterial cell amount information, the bacterial cell amount information is displayed on the display unit 160.

  The conversion information is stored in advance in the storage device 140, but the present invention is not limited to this. The conversion information may be generated by the user and stored in the storage device 140. Thereby, in the subsequent measurement, the generated conversion information can be used for specifying the amount of bacterial cells.

  Specifically, a reference sample containing bacteria having a known bacterial cell amount is dropped into the well of the sample plate 220. After that, under a predetermined measurement condition, the light projecting unit 230 irradiates the reference sample with light, the light receiving unit 240 receives the scattered light, and the intensity acquiring unit D acquires the intensity of the scattered light. The conversion information generation unit H generates conversion information based on the intensity of the scattered light acquired by the intensity acquisition unit D and the known bacterial cell amount, and stores the conversion information in the storage device 140.

  In addition, when two or more types of reference samples having different cell masses are prepared, conversion is performed based on the cell mass in two or more types of reference samples and the intensity of two or more scattered lights respectively corresponding to these. Information may be generated. On the other hand, the inventors of the present invention have confirmed that the intensity of scattered light and the amount of bacterial cells have a proportional relationship. Therefore, only one type of reference sample containing bacteria having a known bacterial cell amount is prepared, and conversion information is generated based on the proportional relationship between the bacterial cell amount in the reference sample and the intensity of scattered light corresponding thereto. Also good.

  The above measurement process may be performed under measurement conditions different from the measurement conditions at the time of generating the conversion information. When the intensity of the emitted light beam by the light projecting unit 230 or the light receiving sensitivity of the light receiving unit 240 changes, the intensity of the scattered light acquired by the intensity acquiring unit D changes in proportion to the intensity of the outgoing light or the light receiving sensitivity. In addition, when the wavelength of the emitted light beam changes, the intensity of the scattered light acquired by the intensity acquisition unit D changes as the quantum efficiency of the light receiving unit 240 changes. In this case, there is a predetermined relationship between the intensity of scattered light and the measurement conditions.

  Therefore, the relationship between the intensity of the scattered light and the measurement conditions is acquired in advance by a preliminary experiment. A correction rule is generated based on the acquired relationship. The user can input measurement conditions in the measurement process to the conversion information correction unit I using the operation unit 150. The conversion information correction unit I corrects (calibrates) the conversion information acquired by the conversion information acquisition unit E based on measurement conditions and correction rules. In this case, the bacterial cell amount specifying unit F specifies the bacterial cell amount in the sample 10 based on the conversion information corrected by the conversion information correcting unit I.

  In the present embodiment, the number of light projecting units 230, the irradiation position of the emitted light beam, the emission direction of the emitted light beam, the diameter of the emitted light beam, and the wavelength of the emitted light beam according to the type, form, and characteristics of the bacteria contained in the sample 10 The measurement conditions including can be arbitrarily changed. Similarly, measurement conditions including the number of light receiving portions 240, the light receiving position, the light receiving direction, and wavelength dependency can be arbitrarily changed according to the type, form, and characteristics of bacteria contained in the sample 10.

  FIG. 3 is a diagram illustrating an example of preferable measurement conditions. In FIG. 3, the sample plate 220 is shown in an enlarged state. As shown in FIG. 3, a plurality of wells 221 are formed in the sample plate 220. Each well 221 has a circular shape with a diameter d1. The sample 10 is dropped into an arbitrary well 221.

  The light projecting unit 230 irradiates the sample 10 in the well 221 with an emitted light beam having a diameter d2. Here, the diameter d2 of the emitted light beam is preferably equal to or larger than the diameter d1 of the well 221. In this case, it is possible to irradiate the entire surface of the sample 10 in each well 221 with the emitted light beam. Thereby, it is possible to reliably and easily irradiate the bacteria contained in the sample 10 with the emitted light beam. Moreover, it is preferable that an emitted light beam contains the wavelength (for example, 337 nm) of an ultraviolet region. Thereby, the light projecting unit 230 can be commonly used as a light projecting unit for ionization of the analysis unit 300 described later. Therefore, the light projecting unit 230 functions as first and second light projecting units.

  Furthermore, it is preferable that the light projecting unit 230 and the light receiving unit 240 are arranged so that the optical axis 231 of the light projecting unit 230 and the optical axis 241 of the light receiving unit 240 are asymmetric with respect to the normal line of the sample plate 220. Accordingly, the light receiving unit 240 can efficiently receive the scattered light from the sample 10 without receiving the light regularly reflected by the sample plate 220.

(3) Analyzer FIG. 4 is a diagram showing a detailed configuration of the analyzer 1 of FIG. 4 is an ion trap mass spectrometer using an ion source based on the MALDI method, and includes an ionization unit 310, an ion trap 320, a mass analyzer 330, a detection unit 340, and an analysis control unit 350. The analyzer 1 may be a mass spectrometer using a method different from the MALDI method, or may be an analyzer other than the mass analyzer. The ionization unit 310, the ion trap 320, the mass analyzer 330, and the detection unit 340 constitute the analysis unit 300.

  2 is provided in the ionization unit 310. The ionization unit 310 includes a sample plate 220 and a light projecting unit 230 that are common to the bacterial cell amount measurement unit 200, and includes an extraction electrode 311. As described above, the microbial cell amount in the sample 10 on the sample plate 220 is measured by the microbial cell amount measuring unit 200. Thereafter, the matrix is mixed with the sample 10 on the sample plate 220.

  The light projecting unit 230 irradiates the sample 10 on the sample plate 220 with pulsed light in the ultraviolet region. Thereby, various components contained in the sample 10 are ionized. The ionization unit 310 may be provided with a light projecting unit separate from the light projecting unit 230 used for the measurement process. The extraction electrode 311 extracts a generated ion toward the ion trap 320 by forming a predetermined electric field.

  The ion trap 320 captures ions extracted from the ionization unit 310 by forming a quadrupole electric field, and cools the ions by emitting a cooling gas to the captured ions. The cooling gas is, for example, helium gas or argon gas. The ion trap 320 releases ions by applying a predetermined electric field to the cooled ions. Ions emitted from the ion trap 320 are introduced into the mass analyzer 330.

  In the present embodiment, the mass analyzer 330 is a time-of-flight mass analyzer. The ions introduced into the mass analyzer 330 fly in the flight space in the mass analyzer 330 at a flight speed corresponding to the mass-to-charge ratio, and reach the detection unit 340 in order from the ions with the lowest mass-to-charge ratio. The detection unit 340 is, for example, a secondary electron multiplier. The detection unit 340 detects ions flying through the mass analyzer 330 and generates a signal corresponding to the detection amount (hereinafter referred to as a detection signal).

  The analysis control unit 350 includes a bacterial cell amount information acquisition unit a, an analysis condition setting unit b, a light projection control unit c, an analysis data processing unit d, and an analysis result output unit e. The functions of the components (a to e) of the analysis control unit 350 are realized by the CPU 110 in FIG. 1 executing the mass spectrometry program stored in the storage device 140. Some or all of the components (ae) of the analysis control unit 350 may be realized by hardware such as an electronic circuit.

  The bacterial cell amount information acquisition unit a acquires the bacterial cell amount information stored in the storage device 140. The analysis condition setting unit b sets analysis conditions based on the bacterial cell amount information acquired by the bacterial cell amount information acquiring unit a. The analysis conditions include, for example, the number of detection signal integrations. The light projecting control unit c controls the operation of the light projecting unit 230 based on the analysis conditions set by the analysis condition setting unit b. Specifically, the light projecting control unit c adjusts the number of times the light is emitted by the light projecting unit 230.

  The analysis data processing unit d generates mass spectrum data indicating a mass spectrum based on the detection signal generated by the detection unit 340. Also, the analysis data processing unit d integrates the generated mass spectrum data the number of times set by the analysis condition setting unit b. The analysis result output unit e outputs the mass spectrum data integrated by the analysis data processing unit d to the storage device 140 and the display unit 160. Thereby, the integrated mass spectrum data is stored in the storage device 140. A mass spectrum based on the mass spectrum data is displayed on the display unit 160.

(4) Measurement process and mass spectrometry process FIG. 5 is a flowchart showing an algorithm of a measurement process performed by the bacterial cell amount measurement program. First, the user designates measurement conditions using the operation unit 150. Thereby, the designated measurement conditions are set in the stage control unit A, the light projection control unit B, and the light reception control unit C.

  The light projection control unit B irradiates the sample 10 with the emitted light beam by the light projection unit 230 (step S1). The intensity acquisition unit D acquires the intensity of scattered light from the sample 10 based on the light reception signal generated by the light receiving unit 240 (step S2). The conversion information acquisition unit E acquires conversion information from the storage device 140 (step S3). Any of Steps S1, S2 and Step S3 may be executed first or at the same time.

  Next, the conversion information correction | amendment part I determines whether correction | amendment of conversion information was instruct | indicated (step S4). The user can instruct correction of conversion information using the operation unit 150. In addition, when the measurement conditions designated by the user are different from the predetermined measurement conditions, correction of conversion information may be automatically instructed. If correction of conversion information is not instructed, the conversion information correction unit I proceeds to step S6. When the conversion information correction is instructed, the conversion information correction unit I corrects the conversion information based on the measurement condition specified by the user (step S5), and proceeds to step S6.

  In step S6, the bacterial cell amount specifying unit F specifies the bacterial cell amount in the sample 10 based on the scattered light intensity acquired in step S2 and the conversion information acquired or corrected in steps S3 and S5. (Step S6). The measurement result output unit G outputs the bacterial cell amount information indicating the bacterial cell amount specified in step S6 to the storage device 140 and the display unit 160 (step S7), and ends the measurement process.

  After the measurement process, the user takes out the sample plate 220 from the analysis unit 300 (the cell amount measurement unit 200), and mixes the matrix with the sample 10 on the sample plate 220. Thereafter, the user sets the sample plate 220 in the analysis unit 300 again. In this state, mass spectrometry processing is executed. Even when the bacterial cell amount measurement unit 200 is provided independently of the analysis unit 300, a common sample plate 220 can be used for the measurement process and the mass spectrometry process.

  FIG. 6 is a flowchart showing an algorithm of mass spectrometry processing performed by the mass spectrometry program. First, the bacterial cell quantity information acquisition unit a acquires the bacterial cell quantity information from the storage device 140 (step S11). The analysis condition setting unit b sets the number of integrations based on the bacterial cell amount information acquired in step S11 (step S12). The light projection control unit c executes the analysis operation based on the number of times set in step S12 (step S13). The analysis operation includes irradiation of the sample 10 with light by the light projecting unit 230.

  The analysis data processing unit d generates mass spectrum data based on the detection signal output by the detection unit 340 by the analysis operation of step S3 (step S14). The analysis data processing unit d integrates the mass spectrum data generated in step S14 (step S15).

  Thereafter, the analysis data processing unit d determines whether or not the cumulative number has reached the number set in step S12 (step S16). If the integration number has not reached the set number, the analysis data processing unit d returns to step S13. Steps S13 to S16 are repeated until the number of integration reaches the set number. When the number of integration reaches the set number, the analysis result output unit e outputs the mass spectrum data integrated in step S15 to the storage device 140 and the display unit 160 (step S17), and ends the mass analysis process. .

(5) Effect In the bacterial cell amount measuring apparatus 2 according to the present embodiment, the surface of the bacteria in the sample 10 is irradiated by irradiating the sample 10 containing the bacteria to be measured held on the sample plate 220 with a light beam. The light scatters. The intensity of scattered light from the sample 10 is acquired, and conversion information indicating the correspondence between the intensity of scattered light and the amount of bacterial cells is acquired. Based on the acquired scattered light intensity and conversion information, the amount of bacterial cells in the sample 10 is specified.

  According to this configuration, even when the amount of the sample 10 is very small, the intensity of scattered light from the sample 10 can be acquired. Thereby, the microbial cell quantity of a trace amount sample can be measured.

  In the mass spectrometry process performed by the analyzer 1, the mass spectrum data is integrated an appropriate number of times based on the amount of cells in the sample 10 measured by the cell amount measuring device 2. Therefore, regardless of the amount of cells in the sample 10, the bacterial component can be analyzed appropriately.

  Further, since the common sample plate 220 can be used for the measurement process and the mass spectrometry process, it is not necessary to move the sample 10 from the sample plate 220 to another sample stage for the mass analysis process after the measurement process. Thereby, the convenience of the analyzer 1 is improved. Furthermore, since the measurement of the amount of bacterial cells in the sample 10 and the analysis of the components of the bacteria in the sample 10 are performed in the same state, the relationship between the amount of bacterial cells in the sample 10 and the analysis result of the sample 10 is accurately grasped. can do.

  Moreover, the 1st light projection part in a measurement process and the 2nd light projection part in a mass spectrometry process are comprised by the common light projection part 230. FIG. Therefore, it is not necessary to provide the first light projecting unit and the second light projecting unit separately. Thereby, the cost of the analyzer 1 can be reduced. Further, the space for installing the light projecting unit in the extraction electrode 311 of the analysis unit 300 can be made compact.

(6) Other Embodiments In the above embodiment, the light projecting unit 230 changes the measurement conditions including the number of installations, the irradiation position of the outgoing light beam, the outgoing direction of the outgoing light beam, the diameter of the outgoing light beam, and the wavelength of the outgoing light beam. Although configured to be possible, the present invention is not limited to this. The measurement conditions of the light projecting unit 230 may be determined in advance. Similarly, the light receiving unit 240 is configured to be able to change the measurement conditions including the number of installations, the light receiving position, the light receiving direction, and the wavelength dependency, but the present invention is not limited to this. The measurement conditions of the light receiving unit 240 may be determined in advance.

  DESCRIPTION OF SYMBOLS 1 ... Analytical apparatus, 2 ... Bacterial cell amount measuring apparatus, 10 ... Sample, 100 ... Control apparatus, 101 ... Bus, 110 ... CPU, 120 ... RAM, 130 ... ROM, 140 ... Storage device, 150 ... Operation part, 160 ... Display unit 170 ... Input / output I / F, 200 ... Bacterial mass measurement unit, 210 ... Moving stage, 220 ... Sample plate, 221 ... Well, 230 ... Light projecting unit, 240 ... Light receiving unit, 250 ... Measurement control unit, DESCRIPTION OF SYMBOLS 300 ... Analysis part, 310 ... Ionization part, 311 ... Extraction electrode, 320 ... Ion trap, 330 ... Mass analyzer, 340 ... Detection part, 350 ... Analysis control part, A ... Stage control part, a ... Acquisition of bacterial cell quantity information , B, c... Projection control unit, b... Analysis condition setting unit, C .. light reception control unit, D... Intensity acquisition unit, d .. analysis data processing unit, E .. conversion information acquisition unit, e. , F ... Bacterial mass specific part, G Measurement result output section, H ... conversion information generation unit, I ... conversion information correction unit

Claims (13)

A cell mass measuring device for measuring the cell mass,
A sample plate for holding a sample containing bacteria to be measured;
A first light projecting unit for irradiating the sample held on the sample plate with a light beam;
An intensity acquisition unit for acquiring the intensity of scattered light from the sample;
A conversion information acquisition unit that acquires conversion information indicating the correspondence between the intensity of scattered light and the amount of bacterial cells;
A bacterial cell amount measuring apparatus comprising: a bacterial cell amount specifying unit that specifies the bacterial cell amount in a sample based on the intensity of scattered light acquired by the intensity acquisition unit and the conversion information acquired by the conversion information acquisition unit . Conversion is performed based on the intensity of scattered light acquired by the intensity acquisition unit and the amount of bacterial cells in the reference sample in a state where a reference sample containing bacteria having a known amount of bacterial cells is held on the sample plate. The bacterial cell amount measurement device according to claim 1, further comprising a conversion information generation unit that generates information. At least one of the number of the first light projecting units, the irradiation position of the emitted light beam by the first light projecting unit, the emission direction of the emitted light beam, the diameter of the emitted light beam, and the wavelength of the emitted light beam is configured to be changeable. The bacterial cell amount measuring device according to claim 1 or 2. The sample plate includes a well for holding a sample;
The diameter of the emitted light beam by said 1st light projection part can be set so that the said well may be included, The microbial cell amount measuring apparatus as described in any one of Claims 1-3. A light receiving unit that receives scattered light from the sample and generates a light reception signal indicating the amount of light received,
The said amount acquisition part is a microbial cell amount measuring apparatus as described in any one of Claims 1-4 which acquires the intensity | strength of scattered light based on the light reception signal produced | generated by the said light-receiving part. The bacterial cell amount measuring device according to claim 5, wherein at least one of the number of the light receiving parts, the light receiving position by the light receiving parts, the light receiving direction, and the wavelength dependence is changeable. The first light projecting unit and the light receiving unit are disposed such that an optical axis of the first light projecting unit and an optical axis of the light receiving unit are asymmetric with respect to a normal to the surface of the sample plate. The bacterial cell amount measuring apparatus according to claim 5 or 6. The conversion information shows the correspondence between the intensity of scattered light and the amount of bacterial cells in a predetermined measurement condition,
The bacterial cell amount measurement apparatus further includes a conversion information correction unit that corrects conversion information acquired by the conversion information acquisition unit based on measurement conditions in the first light projecting unit or the light receiving unit,
The said microbial cell amount specific | specification part specifies the microbial cell amount in a sample based on the intensity | strength of the scattered light acquired by the said intensity | strength acquisition part, and the conversion information correct | amended by the said conversion information correction | amendment part. The microbial cell amount measuring apparatus as described in any one of these. The bacterial cell amount measuring device according to any one of claims 1 to 8,
An analysis device comprising: an analysis unit that analyzes a bacterial component in a sample held on the sample plate of the bacterial cell amount measurement device. A cell mass information acquisition unit for acquiring cell mass information indicating the cell mass in the sample identified by the cell mass identification unit of the cell mass measurement device;
The analysis apparatus according to claim 9, further comprising an analysis condition setting unit that sets an analysis condition of the analysis unit based on the bacterial cell amount information acquired by the bacterial cell amount information acquisition unit. The analysis unit outputs a signal indicating the analysis result,
The analysis apparatus further includes an analysis data processing unit that generates analysis data based on a signal output from the analysis unit and integrates analysis data generated by a plurality of analyzes.
The analysis apparatus according to claim 10, wherein the analysis condition setting unit sets the number of integrations in the analysis data processing unit as the analysis condition. The analysis unit includes a second light projecting unit that irradiates light to the sample held on the sample plate,
The analyzer according to any one of claims 9 to 11, wherein the first light projecting unit and the second light projecting unit of the bacterial cell amount measuring device are configured by a common light projecting unit. A method for measuring the amount of bacterial cells for measuring the amount of bacterial cells,
Irradiating a sample containing bacteria to be measured held on the sample plate by the light projecting unit;
Obtaining the intensity of scattered light from the sample;
Obtaining conversion information indicating the correspondence between the intensity of scattered light and the amount of bacterial cells;
And a step of identifying the amount of bacterial cells in the sample based on the acquired intensity of scattered light and the acquired conversion information.

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