JP2009002710A - Light measuring instrument, and specimen identification dispensing device - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a light measuring instrument and a specimen identification dispensing device, capable of making a specimen dispersed into a sample liquid flowing in an inside of a flow channel flow regularly while recognizing an actual flow condition, and capable of enhancing the measurement reliability of optical information in the specimen. <P>SOLUTION: A user can recognize the actual flow condition, by providing two measuring parts 11a, 11b for measuring the optical information of a transmission light in the specimen, a flow velocity calculating part 12 for calculating a flow velocity of the specimen, a flow velocity graph generating part 13 for generating flow velocity graph information, and a flow condition determining part 14 for determining the flow condition. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to an optical measurement device and a specimen identification and dispensing device. In particular, the specimen dispersed in the sample liquid flowing in the flow path is flowed in an orderly manner while recognizing the actual flow condition, and the specimen is selected by the flow velocity value, thereby improving the measurement reliability of the optical information of the specimen. The present invention relates to an optical measurement apparatus and a specimen identification / dispensing apparatus capable of performing the above.

A liquid in which a specimen such as a cell (microscopic object) such as a cell is dispersed in the liquid flows in the capillary tube, and light from the light source is irradiated onto this liquid stream, so that optical information on the specimen in the liquid stream ( It has been proposed to identify specimens by measuring fluorescence information. After the specimen is identified, the specimen is subjected to ultrasonic vibrations in the dispensing unit to form droplets, giving a charge of, for example, several hundred volts. Then, by applying a voltage of several thousand volts from the deflecting plate, the drop position of each droplet is divided into a plus pole side and a minus pole side, and dispensed into an arbitrary container (well) of the dispensing unit.
Tatsuro Yamashita, Shinichiro Niwa, Cell Engineering Vol. 16, no. 10 p1532-1541, 1997

  When flowing the liquid (sample liquid) in which the specimen is dispersed in the conduit as described above, the sample liquid flows (sample flow) so as to be wrapped in the flow of the sealing liquid (sheath flow) using the sheath flow technique. ) To make the sample solution flow to the center of the pipe. At this time, the pressure of the sample flow and the sheath flow is controlled by the set pressure of the regulator, and the position where the specimen flows in the pipe cross section and the flow rate of the sample flow are adjusted to be constant conditions. Moreover, the measurement parameters detected by the pressure sensor of the sample flow and the sheath flow are fed back, the set pressure by the regulator is adjusted, the pressure of the sample flow and the sheath flow is controlled, and the flow rate of the sample flow becomes a constant condition. It may be adjusted as follows (feedback control).

  However, in the conventional method, since the set pressure is determined from the flow rate of the sample flow and the sheath flow and the measurement parameter, the state (flow condition) in which the specimen flows in the pipeline can be determined only from the measurement parameter. . Therefore, there is a problem that the actual flow condition in the pipe cannot be recognized. For example, when clogging or foreign matter adherence occurs in the pipeline, there is a problem that the flow velocity is different even if the same pressure condition is set.

  Therefore, the present invention has been made to solve the above-described problems, and the sample dispersed in the sample liquid flowing in the flow path is flowed in an orderly manner while recognizing the actual flow condition, and the flow velocity value is determined. An object of the present invention is to provide an optical measurement device and a sample identification / dispensing device that can improve the measurement reliability of optical information of a sample by selecting the sample by the above method.

  The following invention is provided to solve the above-mentioned conventional problems.

  The optical measurement apparatus according to the first aspect of the present invention is a light that measures light information of a specimen by irradiating the specimen, which is a measurement target, dispersed in a sample liquid flowing in a flow path. A measuring device, comprising: a light irradiating unit that irradiates light to the specimen; a light receiving unit that receives the optical information obtained by irradiating the specimen with light; and a plurality of measuring units A flow rate calculation unit that is installed and calculates a flow rate value of the sample based on a difference in measurement time of the optical information measured by the plurality of measurement units with respect to the sample and an interval between the plurality of measurement units; It is characterized by providing.

  The optical measurement apparatus according to the second aspect of the present invention is the optical measurement apparatus according to the first aspect of the present invention, wherein the flow rate value of the specimen calculated by the flow rate calculation unit is obtained from the flow state of the sample liquid. The measurement parameter is determined.

  The optical measurement device according to the third aspect of the present invention arranges the flow velocity values of the specimen calculated by the flow velocity calculation unit in the optical measurement device according to the first or second aspect of the present invention in the order calculated. A flow rate graph generation unit that generates time-series flow rate graph information and outputs the generated flow rate graph information on a display unit is provided.

  An optical measurement apparatus according to a fourth aspect of the present invention is the optical measurement apparatus according to the second or third aspect of the present invention, based on the flow velocity graph information or the measurement parameter including the flow velocity value of the specimen. And a flow state determination unit that determines a flow state of the sample in the flow path and outputs the determined result information to an output unit including the display unit.

  The optical measurement device according to the fifth aspect of the present invention is the optical measurement device according to the third aspect of the present invention, wherein the flow velocity graph generator generates a desired graph region range from all the flow velocity graph information. The flow rate value of the sample acquired by the region specifying unit is calculated by the flow rate calculating unit, the region specifying unit acquiring the flow rate value of the sample within the specified graph region range The flow rate graph information in time series arranged in order is generated, and the generated flow rate graph information is output to a display unit.

  The sample identification and dispensing apparatus according to the first aspect of the present invention is a sample identification and separation unit for collecting a target sample to be sampled from a sample to be measured dispersed in a sample liquid flowing in a flow path. An optical measurement apparatus according to any one of the first to fifth aspects of the present invention, and the specimen identified based on the measurement parameter measured by the optical measurement apparatus, through a nozzle And a dispensing unit that dispenses into a portion to be dispensed.

  The sample identification / dispensing device according to the second aspect of the present invention is the sample identification / dispensing device according to the first aspect of the present invention, wherein the sample identification / dispensing device is within the graph region range designated by the region designation unit of the optical measurement device. The sample corresponding to the flow rate graph information is sorted as the target sample.

  The sample identification and dispensing apparatus according to the third aspect of the present invention is the sample identification and dispensing apparatus according to the first or second aspect of the present invention, wherein the graph region range designated by the region designation unit of the optical measurement device. When the sample corresponding to the flow rate graph information in the sample is collected as the target sample, even if the measured flow rate of the target sample is changed, the target sample has a relationship associated with the measured flow rate. On the basis of this, by calculating the time for dispensing, the target specimen can be dispensed from the nozzle tip of the flow channel to the dispensing target site.

The sample identification and dispensing apparatus according to the fourth aspect of the present invention is slower than the flow velocity at the position where the sample as the measurement object is measured in the sample identification and dispensing apparatus according to the third aspect of the present invention. The specimen identification according to the fifth aspect of the present invention is characterized in that one or a plurality of measured objects can be dispensed from the nozzle tip of the flow path to the dispensing object site after the passage of the flow velocity. In the sample identification and dispensing device according to the fourth aspect of the present invention, the dispensing device has a time for dispensing that is calculated based on the flow rate measured and calculated by the sample that is the measurement object. The measurement object is dispensed into the dispensing target site by switching the positional relationship between the tip of the nozzle and the dispensing target site.

  According to the optical measurement apparatus and the specimen identification and dispensing apparatus of the present invention, the actual flow condition can be recognized by using the flow velocity of each specimen flowing in the flow channel as a measurement parameter. In other words, it is possible to check whether there is an abnormality such as clogging of the pipe line and adhesion of foreign matter. For example, the flow condition of each specimen can be displayed on the display as a graph in time series with respect to the measurement time, or other measurement parameters can be displayed to allow the user to recognize the flow condition. In addition, when an abnormality occurs, the abnormal state can be displayed on the display or output by voice so that the user can recognize the abnormality in a short time and the time for recovery to the normal state can be shortened.

  Further, by making the user recognize the flow condition and stabilizing the flow velocity, it is possible to measure optical information without variation. Therefore, the dispensing operation can be executed with high accuracy.

  In addition, since the light irradiation density varies depending on the position in the flow path, selecting and dispensing specimens with no variation in flow velocity can reduce the dispersion of the dispensed objects, and the optical information can be used again. By measuring, optical information without variation can be measured. Therefore, the dispensing operation can be performed with high accuracy, and the dispensing operation can be performed efficiently.

  An embodiment of the present invention will be described with reference to the drawings. In addition, the embodiment described below is for explanation, and does not limit the scope of the present invention. Accordingly, those skilled in the art can employ embodiments in which each or all of these elements are replaced by equivalents thereof, and these embodiments are also included in the scope of the present invention.

  FIG. 1 is a perspective view showing a preferred embodiment of the optical measuring device of the present invention.

  As shown in FIG. 1, the optical measurement device 10 generates two measurement units 11a and 11b that measure optical information of light transmitted through a specimen, a flow velocity calculation unit 12 that calculates a flow velocity of the specimen, and flow velocity graph information. A flow rate graph generation unit 13 and a flow state determination unit 14 for determining a flow condition are provided. The specimens S and SR are also called test minute objects or samples, and a plurality of specimens S and SR are dispersed in the sample liquid 30. The flow of the sample liquid 30 is called a sample flow 50, and the flow of the sheath liquid 31 that wraps around the sample flow 50 is called a sheath flow 55. Here, the sample S is a target sample to be dispensed, and the sample SR is a non-target sample to be discarded.

  The measuring unit 11a includes a laser (light source) 113a, an irradiation optical fiber 112a, a transmitted light receiving fiber 111a, and a light receiving element (PD) 114a. The measuring unit 11b includes a laser 113b, an irradiation optical fiber 112b, a transmitted light receiving fiber 111b, and a light receiving element (PD) 114b.

  The measurement unit 11a irradiates the sample S and SR passing through the sample flow 50 with the excitation light irradiated from the laser 113a via the irradiation optical fiber 112a and receives the light receiving fiber 111a that receives the transmitted light. And received by the PD 114a. The measurement time information received by the PD 114a is transmitted to the flow velocity calculation unit 12. Similarly, in the measurement unit 11b that is separated from the measurement unit 11a by a predetermined distance H and in the flow channel direction (Z direction), the transmitted light with respect to the specimens S and SR passing through the sample flow 50 is received by the PD 114d and received. The measurement time information is transmitted to the flow velocity calculation unit 12. Accordingly, two pieces of measurement time information measured by the measurement unit 11 a and the measurement unit 11 b for the same specimens S and SR are transmitted to the flow velocity calculation unit 12. The predetermined interval H is not particularly limited, but in the embodiment of FIG. 1, it is about 750 μm, which is the diameter of three optical fibers.

  The sample flow 50 is controlled by the set pressure of the regulator 42, and the pressure is measured by the pressure sensor 44. The pressure measured by the pressure sensor 44 is used as a measurement parameter for determining the flow condition or for feedback control of the pressure of the sample flow 50.

  The sheath flow 55 is controlled by the set pressure of the regulator 41, and the pressure is measured by the pressure sensor 43. The pressure measured by the pressure sensor 43 is used as a measurement parameter for determining the flow condition or used for feedback control of the pressure of the sheath flow 55.

  In addition, the measurement unit 11 a in FIG. 1 receives the fluorescence information and the side scattered light information of the specimens S and SR by the photomultiplier (PMT) 25 through the side light receiving fiber 26. Further, the measurement unit 11 b receives the fluorescence information and the side scattered light information of the specimens S and SR by the PMTs 21, 22, 23, and 24 via the side light receiving fiber 27. The optical information received by the PMTs 21, 22, 23, 24, and 25 is used to identify the specimens S and SR.

  The flow velocity calculation unit 12 in FIG. 1 is based on two pieces of measurement time information measured by the measurement unit 11a and the measurement unit 11b for the same specimens S and SR, and an interval H between the measurement unit 11a and the measurement unit 11b. The flow velocity values of the specimens S and SR are calculated.

  The flow velocity graph generation unit 13 in FIG. 1 generates flow velocity graph information in which the flow velocity values of the specimens S and SR calculated by the flow velocity calculation unit 12 are arranged in the calculated time order, and outputs the generated flow velocity graph information to the display unit 15. To display a flow velocity graph. FIG. 2 is a diagram showing an example of a flow velocity graph. As shown in FIG. 2, the vertical axis indicates the flow velocity, the horizontal axis indicates the order of the specimens S and SR, and the flow velocity state of the specimens S and SR in the sample flow 50 is displayed on the display unit 15 to allow the user to recognize. .

  Further, the flow velocity graph generation unit 13 causes the user to specify a desired graph region range in the flow velocity graph displayed on the display unit 15 via an input unit (not shown), and the specified graph region range. An area designating unit 300 for obtaining a flow velocity value is provided, and flow velocity graph information is generated by rearranging the flow velocity values in the designated region range again in the order of time calculated, and the generated flow velocity graph information is output to the display unit 15. Display the flow rate graph.

  FIG. 3 is a diagram showing an example of area range designation of the flow velocity graph information. FIG. 3A is a diagram showing the flow rate measurement result, and FIG. 3B is a diagram showing the fluorescence measurement result. As shown in FIG. 3A, as a result of designating the graph region range 400 in the flow velocity graph shown in FIG. 2, the specimen in the designated range became a flow velocity graph with a stable flow velocity value. That is, the flow rate variation coefficient Cv decreased from 6.7% to 0.9%. In addition, as shown in FIG. 3B, the variation coefficient Cv of the peak frequency of the fluorescence information measured from the specimen in the specified range is also decreased from 38.2% to 34.0%, and the variation is reduced. As a result.

  The flow state determination unit 14 in FIG. 1 includes flow rate graph information generated by the flow rate graph generation unit 13, measurement parameters (flow rate values calculated by the flow rate calculation unit 12, sample flows 50 measured by the pressure sensors 43 and 44, and The pressure of the sheath flow 55, etc.), various set values, etc. are analyzed to determine the flow condition, and the determined result is output to the display unit 15 to allow the user to recognize the flow condition.

  In the optical measurement device 10 according to the embodiment of the present invention described above, the user can recognize the actual flow condition by calculating the flow velocity of the specimens S and SR and using the calculated flow velocity as a measurement parameter. For example, the flow condition of each specimen can be displayed on the display as a graph in time series with respect to the measurement time, or other measurement parameters can be displayed to allow the user to recognize the flow condition. In addition, when an abnormality occurs, the abnormal state can be displayed on the display or output by voice so that the user can recognize the abnormality in a short time and the time for recovery to the normal state can be shortened.

  Next, an example of the sample identification and dispensing apparatus using the optical measurement device 10 will be described. FIG. 4 is a perspective view showing a preferred embodiment of the specimen identifying and dispensing apparatus of the present invention using the optical measuring device 10.

  As shown in FIG. 4, the sample identification and dispensing device 1000 includes an optical measurement device 10 and a sample dispensing unit 80. As described above, the optical measurement device 10 measures the optical information of the samples S and SR, identifies the samples S and SR, calculates the flow velocity values of the samples S and SR, and determines the flow condition.

  The dispensing unit 80 nozzles a plurality of specimens S identified by the optical measuring device 10 with respect to a well (an example of a container) W at an arbitrary dispensing target position of a culture plate (hereinafter referred to as a plate) 82. The sample SR to be injected is discarded via the nozzle 70, or the sample SR to be discarded is discarded to the waste liquid tank 84 via the nozzle 70. Here, the plate 200 has a plurality of wells W, and the plurality of wells W are arranged in a matrix at a predetermined pitch along the X-axis direction and the Y-axis direction. The pitch interval depends on the shape of the tip of the nozzle 70 and may be such that the sample liquid overflowing from the well W does not enter the adjacent well W when the tip of the nozzle 70 is inserted into the well W. . That is, if the well W is deep, the pitch interval may be narrow, but if the well W is shallow, the pitch interval is made as wide as possible. The diameter of the well W is approximately twice the outer diameter of the nozzle tip 70, so that there is an advantage that the tip of the nozzle 70 can be easily inserted.

  In addition, based on the flow velocity value calculated in the optical measuring device 10, the time (arrival time) for the samples S and SR to reach the tip of the nozzle 70 is calculated, and on the basis of the calculated arrival time, the dispensing unit 80 is calculated. Then, the operation of the nozzle 70 is controlled to execute the dispensing process.

  Thereby, the light information without a dispersion | variation can be measured by making a user recognize a flow condition and stabilizing a flow velocity. Therefore, the dispensing operation can be executed with high accuracy.

  In addition, by dispensing the specimens S and SR with a stable flow velocity selected by the region designating unit 300 of the optical measurement device 10 and measuring the optical information again with the optical measurement device 10, there is no variation in optical information. Can be measured. Therefore, the dispensing operation can be performed with high accuracy, and the dispensing operation can be performed efficiently.

  By the way, this invention is not limited to the said embodiment, A various modified example is employable.

  For example, in FIG. 1, by further providing an audio output unit, a lamp, and the like, the user can be made to quickly recognize the abnormality when the abnormality of the flow condition is determined. In addition, the user can be caused to perform an abnormal state recovery operation.

  In addition, by causing the user to recognize the flow condition, it is possible to avoid occurrence of a serious abnormality.

  The dispensing unit 80 is a plate, but is not limited to a plate, and may be a tube, a dish, or the like.

  In the present invention, the excitation light can also be referred to as measurement light or irradiation light.

  The optical measuring device of the present invention is used in fields requiring examination, analysis and analysis of biopolymers of genes, immune systems, proteins, amino acids, and sugars, such as engineering, food, agriculture, fishery processing, etc. It can be applied to all fields such as fields, hygiene, health, immunity, plague, genetic fields such as heredity, and science fields such as chemistry or biology.

It is a perspective view which shows preferable embodiment of the optical measuring device of this invention. It is the figure which showed an example of the flow velocity graph. It is the figure which showed an example of area | region range specification of the flow velocity graph information. It is a perspective view which shows preferable embodiment of the sample identification dispensing apparatus of this invention using the optical measuring device.

Explanation of symbols

DESCRIPTION OF SYMBOLS 10 Optical measuring device 11a, 11b Measuring part 12 Flow velocity calculation part 13 Flow velocity graph production | generation part 14 Flow state determination part 15 Display part 70 Nozzle 80 Dispensing part 300 Area designation | designated part 1000 Sample identification dispensing apparatus S Specimen (sample)
W Well

Claims (10)

An optical measuring device that irradiates light to a specimen to be measured dispersed in a sample liquid flowing in a flow path and measures optical information of the specimen,
A measuring unit comprising a light irradiating unit for irradiating the sample with light, and a light receiving unit for receiving the optical information obtained by irradiating the sample with light;
A plurality of the measurement units are installed, and the flow velocity value of the sample is calculated based on the difference in measurement time of the optical information measured by the plurality of measurement units with respect to the sample and the interval between the plurality of measurement units. A flow velocity calculation unit for
An optical measuring device comprising:   The optical measurement apparatus according to claim 1, wherein the flow velocity value of the specimen calculated by the flow velocity calculation unit is used as a measurement parameter for determining the flow state of the sample liquid.   A flow rate graph generation unit for generating time-series flow rate graph information in which the flow rate values of the specimen calculated by the flow rate calculation unit are arranged in the calculated order and outputting the generated flow rate graph information on a display unit; The optical measurement device according to claim 1, wherein the optical measurement device is provided.   Based on the flow velocity graph information or the measurement parameter including the flow velocity value of the specimen, the flow state of the specimen in the flow path is determined, and the determined result information is output to an output section including the display section. The optical measurement device according to claim 2, further comprising a flow state determination unit that performs the operation. The flow velocity graph generation unit includes a region designating unit that designates a desired graph region range from all the flow velocity graph information, and acquires the flow velocity value of the specimen within the designated graph region range,
Generates the time-series flow velocity graph information in which the flow velocity values of the specimen acquired by the region designating unit are arranged in the order calculated by the flow velocity calculation unit, and outputs the generated flow velocity graph information to a display unit The optical measuring device according to claim 3. A sample identification and dispensing device for dispensing a target sample to be separated from a sample to be measured dispersed in a sample liquid flowing in a flow path,
The optical measurement device according to any one of claims 1 to 5,
A dispensing unit that dispenses the specimen identified based on the measurement parameters measured by the optical measurement device to a dispensing target site via a nozzle;
A specimen identification and dispensing apparatus comprising:   The sample identification dispensing according to claim 6, wherein the sample corresponding to the flow velocity graph information within the graph region range specified by the region specifying unit of the optical measurement device is sorted as the target sample. apparatus.   When the sample corresponding to the flow velocity graph information in the graph region range specified by the region specifying unit of the optical measurement device is sorted as the target sample, the flow velocity of the measured target sample is changed. In addition, by calculating the time for dispensing based on the relationship associated with the flow velocity at which the target sample is measured, the target sample is dispensed from the nozzle tip of the flow path to the dispensing target site. 8. The specimen identifying and dispensing apparatus according to claim 6, wherein the specimen identifying and dispensing apparatus can perform the specimen identifying and dispensing apparatus.   One or a plurality of measured objects are placed on the portion to be dispensed from the tip of the nozzle of the flow path after a flow rate that is slower than the flow velocity at the position where the sample that is the measurement object is measured. 9. The specimen identifying and dispensing apparatus according to claim 8, wherein dispensing is possible. The measurement target is switched by switching the positional relationship between the nozzle tip of the flow path and the portion to be dispensed with a time for dispensing the sample that is the measurement target based on the measured and calculated flow velocity. 10. The specimen identifying and dispensing apparatus according to claim 9, wherein an object is dispensed to the portion to be dispensed.

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