JP2021128057A - Automatic sampling system of underwater plankton - Google Patents

Abstract

To provide an automatic sampling system of underwater plankton capable of enabling continuously imaging of plankton in seawater with a microscope without manual operation by automatically supplying sample seawater to the field of view of a microscope using a simpler system at a low cost by eliminating disadvantages in a conventional manual manner that a lot of work was required to move and adjust the shooting position when imaging plankton in seawater with a microscope.SOLUTION: In the present invention, sample water is allowed to flow according to the height difference, and the inside of the microchannel is photographed with a microscope only when the flow is stopped. By automatically repeating the passage and rest of the sample water flowing in the microchannel by the operation of the solenoid valve, sample water is automatically supplied and imaged. In addition, the flow rate of sample water is controlled by the time interval of operation of the solenoid valve.SELECTED DRAWING: Figure 1

Description

The present invention relates to an automatic sample sampling device for imaging plankton in seawater.

When imaging plankton in seawater with a microscope, conventionally, it has been necessary to place a sample dropped on a slide or the like under the field of view of the microscope and manually move and adjust the imaging position.

JP-A-6-27014

When imaging plankton in seawater with a microscope, conventionally, it has been necessary to place a sample dropped on a slide or the like under the field of view of the microscope and manually move and adjust the imaging position. Therefore, when the plankton is imaged at a plurality of shooting positions, a large amount of work is required for moving and adjusting the shooting positions.

An object of the present invention is to automatically supply sample water to the observation field of a microscope so that plankton in the sample water can be imaged with a microscope without manual operation. We also thought that it was important to configure it with a simple mechanism so that it could be manufactured at low cost.

An automatic sample sampling device including a supply means for automatically supplying sample water to the observation field of view of a microscope, a control means for controlling the flow rate of sample water, and an imaging means for imaging plankton in sample water with a microscope. ..

The present invention is a device that automatically images plankton in seawater. Compared with the method in which all microscopic examinations are performed manually, there is an effect of significantly reducing the amount of work and an effect of obtaining the same result regardless of the operator. Compared with the conventional invention, the feature is that the configuration of the device is simple.

The figure which shows the apparatus structure of this invention The figure which shows the flow rate control by a control device The figure which shows the time chart of the processing of a control device and an image input device. Diagram showing the function graph used in the arithmetic unit

The device configuration of the present invention is shown in FIG. The sample water container 1, the filter 3, the tube 4, the microchannel 5, and the drainage container 6 constitute a supply means for automatically supplying the sample water to the observation field of the microscope 10, which is an example of the observation means. The sample water container 1 is a container in which a liquid can be injected from the upper part and the outlet of the lower part can be connected to the tube 4. The sample water 2 is seawater collected from the sea as a sample to be imaged. The filter 3 is a filter for filtering dust and the like. The tube 4 is a hollow elongated tube made of a material that can be easily bent or crushed. The micro flow path 5 is a transparent plate having a flow path structure having a depth and width of about several μm to several hundred μm, and can be connected to the tube 4. The width and height of the microchannel shall be larger than the plankton to be observed in order to avoid clogging of the channel. The drainage container 6 is a container for collecting the seawater flowing out from the tube 4. A height difference is provided between the sample water container 1 and the drainage container 6, and the sample water 2 flows inside the tube 4 and the micro flow path 5 due to the height difference.

The image input device 7, the control device 8, the solenoid valve 9, the flow rate measuring device 13, and the display device 14 constitute a control means for controlling the flow rate of the sample water. The image input device 7 is a computer that executes processing related to image input. The control device 8 is a device that controls the operation of the electric device. The solenoid valve 9 is an electric device controlled by the control device 8. The tube 4 is arranged at the operating position of the solenoid valve 9, and the tube 4 is crushed when the solenoid valve 9 operates. The flow rate measuring device 13 is a device that measures the drainage flow rate flowing out of the tube 4 for a certain period of time and inputs the integrated flow rate to the control device 8. The display device 14 is a screen display output destination of the image input device 7, and is a device that displays an abnormal state of the flow rate detected by the control device 8.

The image input device 7, the microscope 10, and the camera 11 constitute an imaging means for imaging plankton in seawater with a microscope. The microscope 10 magnifies and captures the optical input of seawater flowing inside the microchannel 5 installed under the observation area with a lens. The camera 11 can capture the optical input captured by the microscope 10 and convert it into image data. The camera 11 and the image input device 7 are connected by a cable and can transmit and receive signals. The external storage device 12 is a storage device that is a storage destination for an image file created by the image input device 7.

An embodiment of the present invention will be described with reference to FIG. The sample water 2 is injected into the sample water container 1. When the sample water 2 is injected, it is filtered through a mesh filter 3 having a size larger than that of the plankton to be observed to remove dust and the like. Due to the height difference between the sample water container 1 and the drainage container 6, the sample water 2 flows inside the tube 4 and the micro flow path 5.

The image input device 7 issues a start command to the control device 6 during the passage of the sample water, and operates the solenoid valve 9 by the command signal from the control device 8. When the solenoid valve 9 presses the tube 4, the tube 4 is closed, and the sample water 2 inside the tube 4 and the micro flow path 5 comes to rest. With the sample water 2 stationary, the inside of the microchannel 5 is imaged by the camera 11 through the microscope 10. By keeping the sample water 2 stationary, an afterimage in the captured image can be suppressed. The image data of the imaging result is input to the image input device 7 and stored as an image file in the external storage device 12.

The flow rate measurement result of the flow rate measuring device 13 installed in the tube 4 is input to the control device 8 and used for the flow rate control and monitoring of the sample water. The image input device 7 displays the monitoring result on the display device 14.

FIG. 2 shows the control operation of the solenoid valve 9 by the control device 8. The integrated flow rate 101 flowing into the drainage container 6 through the tube 4 is measured by the flow rate measuring device 13. The integrated flow rate 101 is controlled by the operation of the solenoid valve 9.

The difference 103 between the integrated flow rate 101 and the target integrated flow rate 102 measured by the flow rate measuring device 13 is input to the calculator 104 as the integrated flow rate deviation ε, and the output ΔT1 = f (ε) is obtained using the following equation 1. The graph of Equation 1 is shown in FIG.
f (ε) = ΔT1'+ p × [ε / a]… (Equation 1)
However, ΔT1'is the initial value of ΔT1, a is the reference value of deviation, and p is the control coefficient. [] Is a Gaussian symbol indicating the floor function, which is defined by Equation 2 below.
[x] = max {n ∈ Z | n ≤ x}… (Equation 2)
However, Z is a set of integers, n is an arbitrary integer, and x is an arbitrary real number.

The output ΔT1 of the arithmetic unit 104 is input to the PWM 105. PWM105 means Pulse Width Modulation and controls the operating time of the solenoid valve 9. Further, in order to stabilize the output ΔT1 of the arithmetic unit 104, a dead band, a filter or the like is applied to the arithmetic unit 104.

FIG. 3 shows a time chart of processing of the control device 8 and the image input device 7. In the control device 8, the PWM 105 switches between the On state and the Off state by using the execution cycle ΔT1 (201) and the duration of the On state ΔT2 (202) as parameters. The solenoid valve 9 operates during the On state.

A command signal is passed to the image input device 7 at the same time as the PWM 105 is started in the On state of the control device, and the image input process 203 is executed after a certain period of time ΔT3 (205) has elapsed. After the image input process 203 is completed, the image storage process 204 is executed.

As a result of executing a series of processes starting from the PWM 105, an image is taken by the image input process 203 in a state where the flow of the sample water is stopped by the operation of the solenoid valve 9, and then the image data is taken by the image storage process 204. Is saved as an image file. When the PWM 105 is in the Off state, the flow of the sample water is restarted. By repeating a series of operations, a device for automatically supplying sample water and taking an image can be realized.

ΔT1 (201), ΔT2 (202), and ΔT3 (205), which are parameters for processing by the control device 8 and the image input device 7, are shown below. The execution cycle of PWM 105 is ΔT1 second. ΔT1 is adjusted by the arithmetic unit 104 based on the integrated flow rate deviation 103 with the initial state ΔT1 ′ as a reference value. The duration of the ON state of the PWM 105 is ΔT2 seconds. The waiting time from when the solenoid valve operation is turned on to when the image input process is turned on is ΔT3 seconds. ΔT1', ΔT2, and ΔT3 can be manually adjusted as appropriate according to actual operations such as the flow velocity of the sample water and the exposure time of the camera. ΔT1 is automatically adjusted by the flow rate at run time.

The flow rate monitoring unit 106 of FIG. 2 has two functions. The first function is the completion judgment. The total flow rate of the integrated flow rate 101 from the start of water flow is calculated and used as the total flow rate. When the total flow rate exceeds the specified value, it is determined to be completed, and a completion command is issued to the control device 8 and the image input device 7. Upon the completion command, the control device 8 stops the operation of the PWM 105 and a series of control processes, and the image input device 7 stops the image input process 203 and the image storage process 204.

The second function is an alarm display. After the start of water flow, the flow rate is monitored, and if the flow rate is below the specified value for a certain period of time, an alarm is displayed on the display device 14 in FIG. 1 to the effect that the flow is stopped, and in other cases, the alarm display is canceled.

In the image input device 7 of FIG. 1, the total volume of the sample water for imaging is calculated, and the state is displayed on the display device 14. The total volume of the imaged portion can be calculated by defining the volume of sample water per imaging and multiplying by the number of imaging times. The number of times of imaging is equal to the number of observation parts of the sample water, and is equal to the number of times of operation of the solenoid valve 9. That is, even when a plurality of images are continuously imaged while the sample water is stationary by one operation of the solenoid valve 9, the number of imaging times is treated as one. The total volume of the imaged portion can be used for determining the completion of imaging and adjusting the flow rate control of the sample water by the solenoid valve 9.

By using the present invention, it is possible to save an image file in which plankton in seawater is imaged. Image files can be used for various processing by a computer. For example, an image recognition process can be performed by inputting an image file to classify the types of plankton in the image. Furthermore, the density can be calculated by counting plankton for each type. This usage method can be used in fields such as grasping the state of plankton generation during red tide in the ocean, investigating marine ecosystems, and grasping the state of plankton culture.

1 Sample water container 2 Sample water 3 Filter 4 Tube 5 Micro flow path 6 Drainage container 7 Image input device 8 Control device 9 Solenoid valve 10 Microscope 11 Camera 12 External storage device 13 Flow measurement device 14 Display device 101 Integrated flow rate 102 Target integrated flow rate 103 Integrated flow deviation 104 Computing unit 105 PWM (Pulse Width Modulation)
106 Flow rate monitoring unit 201 Solenoid valve operation execution cycle ΔT1 [sec]
202 Solenoid valve operation On state Duration ΔT2 [sec]
203 Image input processing standby time ΔT3 [seconds]

Claims (3)

A supply means for automatically supplying the sample water to the observation field of the microscope, a control means for controlling the flow rate of the sample water supplied by the supply means, and an imaging means for imaging the plankton contained in the sample water with the microscope. An automatic sample sampling device characterized by being provided. The automatic sample sampling device according to claim 1, wherein the control means automatically controls the operating time interval of the solenoid valve in order to adjust the integrated flow rate of the sample water to the target integrated flow rate. A claim that defines the volume of sample water per imaging, calculates the total volume of the imaged portion by multiplying the number of imaging times, and uses the total volume of the imaged portion to control the flow rate of the sample water. Item 2. The automatic sample sampling apparatus according to Item 1 or 2.

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