JP2008136415A - Observation device - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an observation apparatus for improving the precision of an agent irritation observation while controlling a cost. <P>SOLUTION: The observation apparatus comprises a sensor (151) for monitoring the presence of administration to a material to be observed and a recording means (170) for recording an administration timing based on the output of the sensor (151). The labor of a user is reduced by the automatic control of an administration timing in this manner even without using an electrically operated administration apparatus. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to an observation apparatus applicable to observation of drug stimulation such as cultured cells.

  A microscope system has been put into practical use for enlarging and photographing cultured cells growing in a culture medium while controlling the environment of the culture medium in the culture vessel (see Patent Document 1, etc.). In this microscope system, time-lapse imaging is effective in order to visually grasp a gradual time change that occurs in cultured cells (see Patent Document 2 and the like). If this time-lapse imaging interval is set to a short period (for example, 100 msec), it is possible to capture a rapid drug reaction that occurs when drug stimulation is applied to cultured cells.

A large number of image data acquired by time-lapse photography is managed by a computer. When a user performs medication in order to observe a drug reaction, it is desirable that information indicating the timing of medication (medication information) is also managed together with image data.
JP 2005-326495 A JP 2006-220904 A

  However, the task of inputting medication information to the computer is time-consuming, and even if medication information can be entered with a single touch, the user's consciousness cannot be concentrated only on medication during medication, so medication may fail. There is sex. For example, vibration may be applied to the medium, and the cultured cells may float in the medium and be out of the field of view of the objective lens. In order to prevent such a failure, an electric medication apparatus has also been proposed, but the cost is high.

  Therefore, an object of the present invention is to provide an observation apparatus that can improve the accuracy of drug stimulus observation while suppressing costs.

The observation device of the present invention is an observation device for observing a change when a medicine is administered to an object to be observed, a sensor for monitoring the presence or absence of the medicine for the object to be observed, and a dosing timing based on an output of the sensor. And recording means for recording.
In addition, it is desirable to further provide a tube for guiding the medicine to a container that houses the object to be observed.

The sensor may detect a change in pressure of the tube that is a path of the medicine.
The sensor may detect a change in the weight of the object to be observed.
Further, the sensor may detect a change in optical characteristics of the object to be observed caused by the medicine that has been dispensed.
Further, the sensor may detect a change in optical characteristics of the tube that is a path of the medicine.

In addition, any one of the observation apparatuses according to the present invention may further include an imaging unit that performs time-lapse imaging of the object to be observed.
In addition, any one of the observation apparatuses of the present invention may further include a display unit that displays information on the medication timing recorded by the recording unit.

  ADVANTAGE OF THE INVENTION According to this invention, the observation apparatus which can improve the precision of chemical | medical agent stimulus observation is realized, suppressing cost.

[Embodiment]
An embodiment of the present invention will be described. This embodiment is an embodiment of a microscope system.
First, the configuration of the microscope system will be described.
FIG. 1 is a configuration diagram of a microscope system. As shown in FIG. 1, the microscope system includes an apparatus main body 10, a computer 170, a monitor 160, and an input device 180. The apparatus body 10 includes a microscope unit 150, a transmission illumination unit 40, a cooling camera 300, an excitation light source 70, and an optical fiber 7.

The microscope unit 150 includes a stage 23, an objective lens unit 27, a fluorescence filter unit 34, an imaging lens 38, a deflection mirror 452, a field lens 411, and a collector lens 41, and the transmission illumination unit 40. Includes a transmission illumination light source 47, a field lens 44, and a deflection mirror 45.
On the stage 23, a chamber 100 containing a transparent culture vessel 20 is placed. The culture vessel 20 is filled with a medium, and cultured cells labeled with a fluorescent substance are grown in the medium. In order to observe the cultured cells from the outside of the chamber 100, a part a of the bottom surface of the chamber 100 and a part b of the top surface of the chamber 100 are transparent portions. Here, for the sake of simplicity, the case where the upper surface of the culture vessel 20 is opened will be described. However, if necessary, the upper surface of the culture vessel 20 is covered with a lid of the same quality as the culture vessel 20.

A plurality of types of objective lenses are attached to the objective lens unit 27 in the X direction of FIG. 1. When the objective lens unit 27 is driven in the X direction by a mechanism (not shown), the objective lens unit 27 is arranged in the optical path of the apparatus main body 10. The type of objective lens to be switched is switched. This switching is performed under the control of the computer 170.
A plurality of types of filter blocks are mounted on the fluorescent filter unit 34 in the X direction of FIG. 1. When the fluorescent filter unit 34 is driven in the X direction by a mechanism (not shown), it is arranged in the optical path of the apparatus main body 10. The type of filter block to be switched is switched. This switching is also performed under the control of the computer 170.

The computer 170 switches the combination of the type of objective lens arranged in the optical path and the type of filter block arranged in the optical path according to the observation method to be set in the apparatus main body 10. Hereinafter, it is assumed that the observation method of the apparatus main body 10 is switched between phase difference observation and two types of fluorescence observation by this switching.
Of these, between the phase difference observation and the fluorescence observation, both the type of filter block arranged in the optical path and the type of the objective lens arranged in the optical path are different, and between the two types of fluorescence observation, there is no difference in the optical path. Only the type of filter block to be arranged is different. Further, the illumination method is different between the phase difference observation and the fluorescence observation.

  During phase difference observation, the computer 170 turns on the transmission illumination light source 47 to make the optical path of the transmission illumination unit 40 effective, and during fluorescence observation, the optical path of the epi-illumination unit (excitation light source 70, optical fiber 7, collector lens). 41, an optical path passing through the field lens 411, the deflecting mirror 452, the fluorescent filter unit 34, and the objective lens unit 27 in this order) is turned on. The excitation light source 70 is turned off when the transmission illumination light source 47 is turned on, and the transmission illumination light source 47 is turned off when the excitation light source 70 is turned on.

  During phase difference observation, the light emitted from the transmitted illumination light source 47 illuminates the observation point c in the culture vessel 20 through the field lens 44, the deflection mirror 45, and the transparent part b of the chamber 100. The light transmitted through the observation point c reaches the light receiving surface of the cooling camera 300 through the bottom surface of the culture vessel 20, the transparent part a of the chamber 100, the objective lens part 27, the fluorescent filter part 34, and the imaging lens 38, and is observed. A phase difference image of point c is formed. When the cooling camera 300 is driven in this state, a phase difference image is captured and image data is generated. This image data (image data of the phase difference image) is taken into the computer 170.

  During fluorescence observation, the light emitted from the excitation light source 70 is the optical fiber 7, the collector lens 41, the field lens 411, the deflection mirror 452, the fluorescence filter unit 34, the objective lens unit 27, the transparent part a of the chamber 100, and the culture vessel 20. The observation point c in the culture vessel 20 is illuminated through the bottom surface of the. As a result, the fluorescent material present at the observation point c is excited and emits fluorescence. This fluorescence reaches the light receiving surface of the cooling camera 300 via the bottom surface of the culture vessel 20, the transparent part a of the chamber 100, the objective lens part 27, the fluorescent filter part 34, and the imaging lens 38, and a fluorescent image at the observation point c is obtained. Form. When the cooling camera 300 is driven in this state, a fluorescent image is captured and image data is generated. This image data (fluorescent image data) is taken into the computer 170.

The computer 170 controls the XYZ coordinates of the observation point c in the culture vessel 20 by controlling the XY coordinates of the stage 23 and the Z coordinates of the objective lens unit 27.
Further, a humidifier (not shown) is connected to the chamber 100 via a silicon tube (not shown), and the humidity and CO ₂ concentration inside the chamber 100 are controlled in the vicinity of predetermined values, respectively. . In addition, the atmosphere around the chamber 100 is appropriately circulated by a heat exchanger (not shown), whereby the internal temperature of the chamber 100 is also controlled in the vicinity of a predetermined value. The computer 170 monitors the humidity, CO ₂ concentration, and temperature inside the chamber 100 through sensors (not shown). On the other hand, the cooling camera 300 is housed in a housing separate from other parts of the apparatus main body 10, and is maintained at the same level as the outside air temperature of the apparatus main body 10 regardless of the internal temperature of the chamber 100.

The chamber 100 is provided with two glass tubes 119 and 120 as shown in FIG. Each of the glass tubes 119 and 120 penetrates the side wall of the chamber 100 and is fixed in position and posture with respect to the chamber 100.
One end of each of the glass tubes 119 and 120 is located outside the casing in which the apparatus main body 10 is housed, and the other end of each of the glass tubes 119 and 120 is a peripheral portion of the culture medium (from the observation point c) in the culture vessel 20. Soaked in the detached part). These glass tubes 119 and 120 are appropriately piped so as not to obstruct the optical path of the apparatus main body 10.

  When a user gives drug stimulation to the cultured cells in the culture container 20, the medium is discharged from one end of one glass tube 120, and the drug is injected into one end of the other glass tube 119 immediately after that. In this case, the glass tube 119 serves as an injection port for a drug to the medium, and the glass tube 120 serves as a discharge port for the medium. In addition, since pipettes (or syringes) are usually used for injecting drugs and discharging media, the inner diameter of each of the glass tubes 119 and 120 is slightly wider than the outer diameter of the tip of the pipette (or syringe). Set to

As described above, since the positional relationship and the posture relationship between the glass tubes 119, 120 and the chamber 100 are fixed, even if a drug is injected or the medium is discharged through these glass tubes 119, 120, the medium There is little fluctuation and the possibility that the cultured cells will float is low.
Furthermore, a dosing sensor 151 is attached to the surface of the glass tube 119 used as an inlet. The medication sensor 151 is a pressure sensor. When the medicine passes through the hollow portion of the glass tube 119, it reacts at the passage timing to generate a signal and sends it to the computer 170. Therefore, the computer 170 can monitor the presence or absence of medication via the medication sensor 151. Therefore, it is not necessary for the user to grasp the timing (medication timing) at which medication is performed manually or to input the medication timing to the computer 170.

Next, the operation of the computer 170 relating to time-lapse shooting will be described.
An observation program is installed in the computer 170, and the computer 170 operates according to the program. Information input from the user to the computer 170 is assumed to be performed via the input device 180.
FIG. 2 is an operation flowchart of the computer 170. As shown in FIG. 2, first, the computer 170 sets the observation method of the apparatus main body 10 to phase difference observation (precisely, low-magnification phase difference observation), acquires image data of a bird view image in that state, It is displayed on the monitor 160 (step S11). The bird view image is an image of a relatively wide area of the culture vessel 20.

  In acquiring the image data of the bird view image, the computer 170 repeatedly acquires the image data of the phase difference image while moving the observation point c in the culture vessel 20 in the XY direction, and acquires the acquired plurality of image data as one sheet. To the image data of the image. Each piece of image data is so-called tile image image data, and the combined image data is bird view image data.

While observing the bird view image displayed on the monitor 160, the user determines time-lapse shooting conditions (interval, number of rounds, observation point, observation method, etc.).
When a condition is input from the user (YES in step S12), the computer 170 creates a recipe in which the condition is written (step S13). The recipe storage destination is, for example, the hard disk of the computer 170. Hereinafter, the interval written in the recipe, the number of rounds, the observation point, and the observation method are referred to as “designated interval”, “designated round number”, “designated point”, and “designated observation method”, respectively.

Thereafter, when an instruction to start time-lapse shooting is input from the user (step S14), the computer 170 starts time-lapse shooting (step S15).
In time-lapse photography, the computer 170 matches the observation point c of the culture vessel 20 with the designated point, sets the observation method of the apparatus body 10 to the designated observation method, and acquires image data in that state. When there are three designated observation methods, three types of image data are continuously acquired while switching the observation method of the apparatus main body 10 between the three designated observation methods. This completes the first round of shooting.

Thereafter, the computer 170 waits for a specified interval from the shooting start time of the first round, and starts shooting of the second round. The shooting method for the second round is the same as that for the first round.
Further, the above photographing is repeated until the number of executed rounds reaches the designated number of rounds (until YES in step S18).

Now, during the time-lapse shooting period (during step S18 NO), the computer 170 sequentially stores the image data captured from the apparatus main body 10 in the execution progress file as shown in FIG. The storage location of the implementation progress file is, for example, the hard disk of the computer 170.
Further, during the time lapse photography period (during step S18 NO), the computer 170 monitors the environment of the culture vessel 20 (humidity, CO ₂ concentration, temperature inside the chamber 100) and information about the environment (environment information). ) Is written in the header of the implementation progress file (see FIG. 3).

Further, during the time lapse imaging period (during step S18 NO), the computer 170 monitors the presence / absence of medication for the culture container 20 (step S16), and if there is medication (step S16 YES), the timing of medication Is written in the header of the implementation progress file (see FIG. 3) (step S17).
Furthermore, when a confirmation instruction is input from the user during the time lapse shooting period or after the end of the time lapse shooting, the computer 170 refers to the contents of the execution progress file at that time, and based on this, the progress check screen described below Is displayed on the monitor 160.

FIG. 4 is a diagram showing an implementation progress confirmation screen. As shown in FIG. 4, display areas 101, 102, 103, 104 and a playback control unit 105 are arranged on the execution progress confirmation screen.
The display area 101 is an area where a moving image of the phase difference image is to be displayed, and the display area 102 is an area where one of the two types of fluorescent images (first fluorescent image) is to be displayed. The display area 103 is an area in which the moving image of the other of the two types of fluorescent images (second fluorescent image) is to be displayed. The display area 104 is an area in which a moving image of a composite image of the phase difference image and the fluorescence image is to be displayed.

  Prior to the display of the moving image, the computer 170 reads the three types of image data acquired in each round from the execution progress file (see FIG. 3), and concatenates each frame of the phase difference image data in time series. To create a moving image file of the phase difference image, connect the frames of the image data of the first fluorescence image in time series, create a moving image file of the first fluorescence image, and A moving image file of the second fluorescent image is created by connecting the frames of the image data in chronological order. Further, the computer 170 synthesizes the image data of the phase difference image and the image data of the fluorescent image for each frame, and creates a moving image file of the synthesized image by connecting the synthesized frames in time series. The storage destination of these four types of moving image files is, for example, the hard disk of the computer 170.

  When displaying moving images, the computer 170 reads the created four types of moving image files simultaneously and in parallel, and displays the four types of moving images on the four display areas 101, 102, 103, and 104 individually. A moving image signal is generated and sent to the monitor 160 in the order of generation. Hereinafter, a series of processing of the computer 170 including reading of the moving image file, generation of the moving image signal, and transmission of the moving image signal is referred to as “reproduction of moving image file”.

The reproduction control unit 105 shown in FIG. 4 is a GUI image for the user to input an instruction regarding reproduction of a moving image file to the computer 170.
FIG. 5 is an enlarged view of the playback control unit 105. As shown in FIG. 5, the playback control unit 105 includes a stop button 52, a skip button 53, a playback button 54, a fast-forward button 55, a selection playback button 56, a section designation button 57, a timeline 50, and the like.

When the user selects the playback button 54, the playback of the moving image file is started, and the display of the moving image on the display areas 101, 102, 103, 104 is started. When the user selects the stop button 52, the moving images displayed in the display areas 101, 102, 103, and 104 are stopped.
The playback location in the moving image file is reflected in the timeline 50. The left end (1) of the time line 50 indicates the beginning of the moving image file (time lapse shooting start time), and the right end (2) of the time line 50 indicates the tail of the moving image file (time lapse shooting end time). When time-lapse shooting is not completed, the right end (2) of the timeline 50 indicates the current time. The slurder bar 60 is arranged on the timeline 50, and the playback position in the moving image file is represented in real time by the position of the slurder bar 60 in the horizontal direction.

The position of the slurder bar 60 in the left-right direction can be freely changed by the user. When the horizontal position of the slurder bar 60 changes, the playback location in the moving image file also changes accordingly.
In addition, on the timeline 50, medication marks 51-1, 51-2, 51-3,... Each medication mark 51-1, 51-2, 51-3,... Is an icon of a medication device such as a pipette or a syringe. Each of the arrangement positions of the medication marks 51-1, 51-2, 51-3,... Indicates the medication timing that occurred during the time-lapse imaging. Prior to the display of these medication marks 51-1, 51-2, 51-3,..., The computer 170 refers to the header of the execution progress file (see FIG. 3), and all medications generated during the time-lapse imaging period. Recognize timing.

When the user selects the selection / reproduction button 56 in this state, the slurder bar 60 jumps to the medication mark (the medication mark 51-3 in FIG. 5) adjacent to the right side thereof. Then, the computer 170 reproduces a predetermined section (ten or more frames) before and after the medication mark 51-3. As a result, the user can observe the drug reaction of the cultured cells with one touch.
If the medication mark is not arranged on the right side of the slurder bar 60 when the user selects the selection reproduction button 56, the jump destination of the slurder bar 60 is the arrangement of the medication mark adjacent to the left side of the slurder bar 60. The playback section is a predetermined section (ten frames or more) before and after the medication mark.

In addition, it is desirable that the number of frames in the playback section when the user selects the selection playback button 56 can be set in advance by the user.
Further, as shown in FIG. 5, it is desirable that the selection reproduction button 56 is given the same mark as the medication marks 51-1, 51-2, 51-3,.
As described above, the microscope system of the present embodiment includes the medication sensor 151 (see FIG. 1), and the computer 170 automatically manages the medication timing using the output of the medication sensor 151. Therefore, the user's consciousness at the time of medication can be concentrated only on medication.

In addition, since the microscope system of the present embodiment is equipped with the glass tubes 119 and 120 for medication, the possibility that the user will fail in medication can be further reduced.
[First Modification]
In the microscope system of the above-described embodiment, the attachment destination of the dosing sensor 151 is the glass tube 119. However, even if the dosing sensor 151 is changed between the position indicated by the reference symbol A or B in FIG. Good. In this case, a change in the weight of the chamber 100 (that is, a change in the weight of the culture vessel 20) is detected. In addition, it is desirable to form a recess for providing the dosing sensor 151 on the surface of the stage 23 so that the posture of the chamber 100 does not become unstable. Also, it is desirable that the timing at which the medication sensor 151 generates a signal is limited only when the weight of the chamber 100 increases.

[Second Modification]
In the microscope system of the above-described embodiment, the medication sensor 151 including the pressure sensor is used. However, a medication sensor 151 as illustrated in FIG. 7 may be used. In FIG. 7, reference numeral 151A indicates a light projecting / receiving unit of the dosing sensor 151, reference numeral 151B indicates an indicator of the dosing sensor 151, and reference numeral 26 indicates an objective lens in use.

[Third Modification]
In the microscope system of the above-described embodiment, the medication sensor 151 including the pressure sensor is used. However, a medication sensor 151 as illustrated in FIG. 8 may be used. In FIG. 8, reference numeral 151 </ b> D indicates a light projecting unit of the medication sensor 151, and reference numeral 151 </ b> C indicates a light receiving unit of the medication sensor 151. The light projecting unit 151D and the light receiving unit 151C of the medication sensor are disposed to face each other with the glass tube 119 interposed therebetween.

  The light projecting unit 151D of the medication sensor 151 emits detection light that does not affect the medicine, and the detection light enters the light receiving unit 151C through the side wall of the glass tube 119, the hollow portion of the glass tube 119, and the side wall of the glass tube 119. To do. If there is no medicine in the hollow part of the glass tube 119, most of the detection light is incident on the light receiving part 151C. However, if there is a medicine in the hollow part of the glass tube 119, the amount of detection light incident on the light receiving part 151C is Lower. The light receiving unit 151C of the medication sensor 151 monitors the amount of incident light and reacts to generate a signal when the amount of light becomes less than a threshold. Such a dosing sensor 151 has the same function as the dosing sensor 151 shown in FIG.

[Others]
In the microscope system of the embodiment or the modification described above, the glass tubes 119 and 120 are used for injecting / extracting the drug to / from the culture container 20, but a tubular member made of another material that does not react with the drug may be used. Good.
However, in the third modified example (see FIG. 8), since the presence or absence of the drug present in the hollow portion of the tubular member is detected from the outside of the tubular member, the material of the tubular member must be at least transparent.

It is a block diagram of a microscope system. 6 is an operation flowchart of the computer 170. It is a figure explaining an implementation progress file. It is a figure which shows the screen for implementation progress confirmation. 3 is an enlarged view of a playback control unit 105. FIG. It is a figure which shows a 1st modification. It is a figure which shows a 2nd modification. It is a figure which shows a 3rd modification.

Explanation of symbols

DESCRIPTION OF SYMBOLS 10 ... Apparatus main body, 170 ... Computer, 160 ... Monitor, 180 ... Input device, 150 ... Microscope part, 40 ... Transmission illumination part, 300 ... Cooling camera, 70 ... Light source for excitation, 7 ... Optical fiber, 119, 120 ... Glass Tube, 151 ... Dosing sensor

Claims (8)

In an observation device for observing changes when a drug is administered to an object to be observed,
A sensor for monitoring the presence or absence of the medication for the object to be observed;
An observation apparatus comprising: recording means for recording medication timing based on the output of the sensor. The observation apparatus according to claim 1,
An observation apparatus, further comprising a tube for guiding the medicine to a container that accommodates the object to be observed. The observation apparatus according to claim 2,
The sensor is
An observation apparatus that detects a pressure change of the tube that is a path of the medicine. In the observation device according to claim 1 or 2,
The sensor is
An observation apparatus for detecting a change in weight of the object to be observed. In the observation device according to claim 1 or 2,
The sensor is
A change in optical characteristics of the object to be observed caused by the dispensed medicine is detected. The observation apparatus according to claim 2,
The sensor is
A change in optical properties of the tube, which is a path of the medicine, is detected. In the observation apparatus as described in any one of Claims 1-6,
An observation apparatus further comprising an imaging unit that performs time-lapse imaging of the object to be observed. In the observation apparatus according to any one of claims 1 to 7,
An observation apparatus, further comprising display means for displaying information on medication timing recorded by the recording means.

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