WO2005031322A1 - 環境保持装置および環境制御型分析装置 - Google Patents
環境保持装置および環境制御型分析装置 Download PDFInfo
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- WO2005031322A1 WO2005031322A1 PCT/JP2004/013539 JP2004013539W WO2005031322A1 WO 2005031322 A1 WO2005031322 A1 WO 2005031322A1 JP 2004013539 W JP2004013539 W JP 2004013539W WO 2005031322 A1 WO2005031322 A1 WO 2005031322A1
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- chamber
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- environment
- environmental
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Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/27—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands using photo-electric detection ; circuits for computing concentration
Definitions
- the present invention relates to an environment control type analyzer for holding, observing, and analyzing a sample, particularly, a biological sample.
- an environment holding device includes a first chamber that holds a sample and has an open surface, and a moving stage that mounts the first chamber and moves two-dimensionally.
- a first surface provided to cover an open surface of the first chamber, the first chamber having an introduction portion for introducing gas under predetermined conditions into the first chamber via a pipe.
- the first surface is provided so as to be fixed against the movement of the moving stage, and the open surface of the first chamber is provided with a moving surface.
- the tage moves two-dimensionally along the first plane.
- the moving stage moves within a range where the position of the introduction portion of the first surface is located within the open surface. Is preferred.
- the predetermined condition of the gas preferably includes at least one of temperature, humidity, and composition.
- the gas composition preferably includes air and carbon dioxide gas.
- the first surface has an opening / closing portion that can be partially opened / closed.
- the opening / closing section includes a first state in which the first chamber is opened to the outside through an opening, and a second state in which the opening is closed. It is preferable to be able to switch between the states.
- the opening / closing section includes a first state in which the first chamber is opened to the outside through an opening, and a second state in which the opening is closed. It is preferable that the state can be switched between the state described above and the third state in which illumination light is introduced into the first chamber through a transmission window member.
- the apparatus further comprises a second chamber for accommodating the first chamber and the moving stage,
- the surface is preferably one of the surfaces constituting the outer shape of the second chamber.
- the first chamber has a first opening and a shirt that opens and closes the first opening. It is preferable that the first chamber has a second opening and a shutter for opening and closing the second opening at a position facing the first opening.
- the environmental control type analyzer is any one of the first to tenth aspects.
- An environment holding device of the embodiment an analyzer for analyzing a sample held in the first chamber, and a pipe and a first surface for holding the inside of the first chamber in a predetermined environment.
- An environment control device connected to the first chamber.
- an environmental control type analyzer comprises: an environment holding device according to the eighth aspect; an analyzer for analyzing a sample held in the first chamber; A transmission illuminator, which is provided to face the detector of the analyzer and illuminates the sample through a window member, and a pipe and a first surface for maintaining the inside of the first chamber in a predetermined environment. And an environment control device connected to the first chamber through the first chamber.
- an environmental control type analyzer comprises the environment maintaining device of any one of the ninth to tenth aspects, at least a part disposed in the second chamber, and a first chamber.
- the environmental control type analyzer of the thirteenth aspect preferably further comprises a dehumidifier for dehumidifying the inside of the second chamber.
- the present invention which is configured as described above, provides an environment-holding device that can move a sample without moving a pipe and an environment-controlled analyzer that can analyze each part of a sample without moving a pipe. Can be provided.
- FIG. 1 is a configuration diagram schematically showing a configuration of an environment holding device according to an embodiment of the present invention.
- FIG. 2 is a top view of the environment holding device according to the embodiment of the present invention.
- FIG. 3 is a cross-sectional view taken along the line II of FIG. 2, and is a diagram illustrating a configuration of a shutter mechanism.
- FIG. 4 is a configuration diagram for explaining the operation of the environment preservation device according to the embodiment of the present invention.
- FIG. 5 is a configuration diagram schematically showing the entire configuration of the environment-controlled analyzer according to the embodiment of the present invention, in which the sample chamber is set at a reference position.
- FIG. 6 is a configuration diagram schematically showing the entire configuration of the environment-controlled analyzer according to the embodiment of the present invention, in which the sample chamber is set at a position shifted from the reference position.
- FIG. 7 is a configuration diagram schematically showing an overall configuration of an environment control type analysis device using an erect microscope, which is a modification of the embodiment of the present invention.
- FIG. 1 is a configuration diagram schematically showing a configuration of an environment control type analyzer according to the present embodiment.
- FIG. 2 is a top view of the environment-controlled analyzer according to the present embodiment.
- FIG. 3 is a cross-sectional view taken along the line II of FIG. 2, and is a diagram illustrating a configuration of the shirt clutch mechanism.
- FIG. 4 is a configuration diagram for explaining the operation of the environment control type analyzer according to the present embodiment.
- the same components are denoted by the same reference numerals. For convenience of explanation, directions are represented by X, Y, Z rectangular coordinates as shown in the figure.
- the environment control type analyzer includes an environment holding device 1 for holding a sample in a certain environment, a microscope device 30 for analyzing a sample, and an environment for controlling the environment in the environment holding device 1. It comprises a control (adjustment) device 50, a control unit 24 for controlling these devices, a PC 25, and the like.
- the present invention is particularly characterized by the above-mentioned environment preserving device 1.
- an environment preservation apparatus 1 of the present embodiment has a configuration in which a sample chamber 10 as a first chamber is housed in a closed container 20 as a second chamber. Become. The inside of the sample chamber 10 is maintained in a predetermined environment state by the environment control device 50. This environmental control device 50 will be described later with reference to FIGS.
- the sample chamber 10 is opened such that one surface of a ceiling portion is indicated by an open surface 10A, and an opening 11A is formed in the bottom plate 11.
- the culture vessel 12 holding the biological sample S is placed on the bottom plate 11 so as to close the opening 11A.
- the sample chamber 10 is mounted on an XY moving stage 22, and moves in the X and Y directions along a horizontal plane.
- an opening / closing window 13 is provided on a side surface of the sample chamber 10 for inserting and removing the culture container 12 and for performing maintenance inside the sample chamber.
- the closed container 20 includes a sample chamber 10, an XY moving stage 22, and a microscope device serving as an analyzer. This is a container for storing a part of the device 30.
- the open surface 10A of the sample chamber 10 is in parallel and close to the lower surface 21A of the upper plate 21, which is one of the walls constituting the outer shape of the closed vessel 20. Even when the sample chamber 10 moves in the XZY direction along the horizontal plane, the open surface 10A of the sample chamber 10 and the lower surface 21A of the upper plate 21 of the closed container 20 maintain a close parallel state. That is, while maintaining the state in which the end of the side wall surface (such as the side plate 15 in FIG. 5) of the sample chamber 10 forming the open surface 10A is close to the lower surface 21A of the upper plate 21 of the sealed container 20, 10 moves in parallel. Further, the upper plate 21 of the closed container 20 covers the open surface 10A.
- the upper plate 21 of the closed vessel 20 almost completely closes the open surface 10A of the sample chamber 10, and the inside of the sample chamber 10 and the inside of the closed vessel 20 are always independently airtight. Will be retained.
- the sealed container 20 is fixed to the microscope device 30 so that the sample chamber 10 does not move even if the sample chamber 10 moves in the XZY direction by the XY moving stage 22. That is, the upper plate 21 is provided so as to be fixed with respect to the movement of the XY movement stage 22.
- An opening / closing window 23 is provided on the side of the sealed container 20 for taking the culture container 12 in and out, and for maintaining the inside of the sample chamber and the sealed container.
- the upper plate 21 has a shutter mechanism described later. 40 are provided.
- the microscope device 30 has an objective lens 31, a focusing device 32, an excitation light illuminating device 33, a fluorescent filter device 34, a second objective lens 35, a reflecting prism 36, an imaging device 37, and a transmission illuminating device 38.
- the objective lens 31 is held by the focusing device 32 and moves in the direction of the optical axis AX, that is, in the Z direction.
- the transmitted illumination device 38 is arranged above the closed container 20.
- the control unit 24 is electrically connected to the microscope device 30, the shutter mechanism 40, and the XY movement stage 22 via wiring.
- the control unit 24 is connected to a personal computer (PC) 25.
- the control unit 24 acquires various data relating to observation conditions, photographing conditions, stage movement conditions, and the like from the PC 25, and outputs them as control signals to the microscope device 30, the shutter mechanism 40 that is an opening / closing means, and the XY movement stage 22. Further, the control unit 24 outputs various control data and image data to the PC 25.
- the shutter mechanism 40 includes a disk-shaped wheel 41, a rotating shaft 42, a gear 43, a motor 44, and a cover 45.
- the wheel 41 is mounted on the rotating shaft 42, the gear 43 is mounted on the rotating shaft of the motor 44, and the wheel 41 and the gear 43 are connected to each other. I'm in love.
- the wheel 41 is provided with an opening 41a for introducing a biological sample into the first chamber, and a transparent substrate 4 lb as a transmission window member for introducing illumination light into the first chamber.
- Reference numeral 41c which is indicated by an imaginary line, is a light-shielding region having no opening or transparent substrate.
- the wheel 41, the rotating shaft 42, the gear 43, and the motor 44 are covered by a cover 45 having an opening 45A.
- the wheel 41 has a first position that opens the first chamber to the outside through an opening 41a, a second position that closes the opening 41a, and a transmission window member 41b inside the first chamber. It can be switched to the third position for introducing illumination light. That is, the shutter mechanism 40 includes a first state in which the first chamber is opened to the outside through the opening 41a, a second state in which the opening 41a is closed, and a state in which the first chamber is transmitted through the first chamber. The state is switched to the third state in which the illumination light is introduced through the window member 41b.
- FIG. 4 the electrical wiring between the control unit 24 and the PC 25 shown in FIG. 1 is omitted.
- the sample well plate 14 (corresponding to the culture vessel 12 in FIG. 1) is set in the sample chamber 10 through the opening / closing window 23 and the opening / closing window 13 sequentially.
- Each of the opening / closing window 23 and the opening / closing window 13 has a structure in which a shirt is provided at an opening.
- the well plate 14 is a transparent container in which a plurality of wells are arranged.
- the signal from the control unit 24 causes the motor 44 of the shutter mechanism 40 to rotate, which in turn rotates the gear 43, which rotates the wheel 41, and the center of the opening 41a is positioned on the optical axis AX. .
- the inside of the sample chamber 10 is open to the outside world.
- the pipette 47 of the injector 46 is inserted into the opening 4 la, and the tip of the pipette 47 reaches the sample chamber 10.
- the injector 46 can be moved in the X, ⁇ , and Z directions by a drive mechanism (not shown) in response to a signal from the control unit 24, and a culture solution or a reaction reagent containing a biological sample can be transferred via a pipette 47. It is configured so that it can be injected into the well of the plate 14.
- the well plate 14 is moved in the XZY direction by the XY moving stage 22.
- the transmitted illumination device 38 also loses its force on the optical axis AX as shown in the figure.
- the pipette 47 of the injector 46 retreats upward, and the signal from the control unit 24 activates the shutter mechanism 40 to replace the opening 41a with the transparent substrate 41b or light shielding.
- the area 41c is located on the optical axis AX.
- the transparent substrate 4 lb is located on the optical axis AX
- the light shielding area 41c is located on the optical axis AX.
- the transmission illumination device 38 returns to the optical axis AX, and the illumination light irradiates the biological sample S.
- the light transmitted through the biological sample S is incident on the objective lens 31 via the well plate 14.
- the light that has entered the objective lens 31 passes through the fluorescent filter device 34 and the second objective lens 35, is reflected by the reflection prism 36, and forms an image on the imaging device of the imaging device 37.
- observation conditions such as brightness adjustment of the illumination light source, switching of various filters, switching of the observation magnification, and adjustment of the field stop are set by the control signal from the control unit 24.
- shooting conditions such as CCD gain, shutter speed, numerical aperture, and shooting timing in conjunction with a lighting device are set by a control signal from the control unit 24.
- the microscope image data of the biological sample S is sent to the PC 25 via the control unit 24, and is displayed on the display as a microscope image.
- the PC 25 can also process and display a microscope image.
- the PC 25 can also display control data such as the above-mentioned observation conditions and imaging conditions on a display as needed.
- the well plate 14 When observing a biological sample S injected into another well, the well plate 14 is moved in the X / Y direction by the XY moving stage 22 according to a control signal from the control unit 24. ⁇ If the focal position shifts due to the movement of the L-plate 14 or switching of the observation magnification, the objective lens 31 can be moved in the Z direction by the focusing device 32 to adjust the focus on the biological sample S . Control data such as stage movement conditions can also be displayed on the display of the PC 25 as needed. For example, if a numerical value is displayed to indicate how far the XY movement stage 22 is from the reference position, the biological sample injected into each well can be displayed. The fee s can be easily identified.
- light emitted from the excitation light illuminating device 33 passes through a dimming filter (not shown) and a fluorescent filter device 34 and is incident on a lower portion of the objective lens 31.
- the light incident on the objective lens 31 is radiated through the well plate 14 onto the biological sample S to which the fluorescent substance has been added.
- the excitation light causes the biological sample S to emit fluorescence.
- the fluorescent light passes through the mirror plate 14, the objective lens 31, the fluorescent filter device 34, and the second objective lens 35, is reflected by the reflection prism 36, and forms an image on the imaging device of the imaging device 37.
- Microscopic observation, image data processing, stage movement, etc. can be basically the same for transmission image observation and fluorescence image observation.
- FIG. 5 is a configuration diagram schematically showing the overall configuration of an environment control type analyzer composed of the environment holding device 1, the environment control device 50, and the microscope device 30 shown in FIG. 1.
- the sample chamber is located at the reference position. Is set.
- FIG. 6 is a configuration diagram schematically showing the entire configuration of the environmental control type analyzer, in which the sample chamber is set at a position where the reference position force has also moved.
- FIGS. 5 and 6 are diagrams each showing a state in which the environment control device 50 is connected to the environment holding device 1 shown in FIG. 1, and the same components as those in FIGS. The description is omitted.
- the illustration of the control unit 24 and the PC 25 is omitted.
- the environment control device 50 is a device that generates a gas having a desired temperature, humidity, and composition, and circulates the gas into the sample chamber 10. That is, a gas (gas) set under predetermined conditions is sent into the sample chamber 10.
- the environmental control device 50 includes a humidifier 51, a heater 52, a blower 53, a temperature Z humidity sensor 54, a CO gas sensor 55, a reserve tank 61, and a cooler 62.
- the heater 52 is connected to the upper plate 21 of the sealed container 20 by a gas delivery tube 56.
- the cooler 62 is disposed in the humidifier 51, and is provided with the gas suction tube 57a and the CO gas
- a pipe is connected to the upper plate 21 of the sealed container 20 by a gas suction tube 57b in which a sensor 55 is inserted.
- the pipe connection point of the upper plate 21 communicates with the sample chamber 10.
- the caro humidifier 51 is connected to the dehumidifier 60 via a drain tube 58 by piping.
- a solenoid valve 59 is provided in the path where the force between the heater 52 and the blower 53 also branches, and a CO gas cylinder 70
- the temperature Z humidity sensor 54 is attached to the lower surface of the upper plate 21 of the sealed container 20 to monitor the temperature and humidity in the sample chamber 10.
- the CO gas sensor 55 is located inside the sample chamber 10.
- Each tube Being beaten.
- Each tube is heat-insulated.
- the gas supplied to the sample chamber 10 by the gas delivery tube 56 is adjusted in gas composition, humidity and temperature as follows. Adjust the gas composition by measuring with the CO gas sensor 55
- the adjustment of the humidity is performed by feeding back the measured value of the temperature Z humidity sensor 54 and increasing or decreasing the drive voltage to the ultrasonic atomizing element of the humidifier 51.
- the hydraulic power in the humidifier 51 also generates water vapor by ultrasonic vibration. Note that a constant amount of water in the humidifier 51 is always ensured by the supply from the reserve tank 61.
- the temperature is adjusted by feeding back the measured value of the temperature Z humidity sensor 54 and increasing or decreasing the current flowing to the electric heater of the heater 52. Further, the temperature of the gas supplied to the sample chamber 10 can be adjusted by using the heater 52 and the cooler 62 together.
- the gas adjusted in this manner is, for example, 37 ° C-100% RH, 5% CO,
- the gas is supplied from the control device 50 to the sample chamber 10 by the gas delivery tube 56, and is returned from the sample chamber 10 to the environmental control device 50 by the gas suction tubes 57a and 57b.
- a very small amount of the adjusting gas is supplied to the closed container 20 through the gap between the upper plate 21 of the closed container 20 and the side plate 15 of the sample chamber 10 or the gap between the plate 14 and the bottom plate 16 of the sample chamber 10.
- the leaked adjustment gas is separated into moisture by the dehumidifier 60 and the water is discharged to the humidifier 51 by the drain tube 58.
- the inside of the closed container 20 is dried at 37 ° C and 5% CO, for example. Environment is maintained.
- a Peltier device may be used as the dehumidifier 60.
- Silica gel can be used as a simple method.
- the conditioning gas on the circulation path is sent again into the sample chamber 10 by the environment control device 50 as air having a predetermined temperature, humidity, and gas composition.
- the inside of the sample chamber 10 is always maintained in a predetermined environment, and the observation and analysis of the biological sample S can be performed. Since the sample chamber 10 is housed in the sealed container 20, there is no danger of the adjustment gas leaking to the outside!
- an environment-controlled analyzer according to the present embodiment when the sample chamber 10 is moved from the reference position will be described with reference to FIG. In FIG. 6, when the reference position is the optical axis AX of the objective lens, the sample chamber 10 is located on the left side by a distance d from the optical axis AX.
- the mounting positions are as follows. Pl, P2, P3, and P4 are all in the range of length L. That is, the sample chamber 10 moves within a range where the mounting positions Pl, P2, P3, and P4 are located within the open surface 10A of the sample chamber 10 (Fig. 1).
- the circulation path through which the adjustment gas flows remains stationary even during the movement. Therefore, the inside of the sample chamber 10 is always kept in a predetermined environment, and the observation and analysis of the biological sample S can be stably performed for a long time. Further, in the apparatus of this embodiment, since only the sample chamber 10 in the closed vessel 20 moves, the wiring of the XY movement stage device 22 and the shutter mechanism 40 is immovable, and there is no danger of disconnection! / Piping and wiring do not become driving resistance of the driving device.
- a reagent or a culture solution may be added to the sample.
- the addition of such reagents and culture medium is performed by the injector 46 and the pipette 47.
- the size of the opening is desirably small because the opening communicates with the outside air.
- a reagent or the like can be injected into each well by moving the first chamber 10. That is, even when a reagent or the like is injected into a wide range of samples, it is not necessary to provide a large opening corresponding to a wide range of samples.
- the microscope apparatus 30 has been described as an analysis apparatus.
- a photosensor may be provided instead of a microscope to measure the intensity and saturation of the fluorescence from the biological sample S.
- other analyzers may be used. That is, the present invention can be applied to any analyzer that analyzes a sample while holding the sample in a certain environment.
- the entire analyzer may be housed in the closed container 20, or only a part of the analyzer may be housed.
- optical members from the objective lens 31 of the microscope device 30 to the fluorescent filter device 34 may be housed in the closed container 20. According to this configuration, the internal volume of the closed container 20 can be reduced, and compactness can be achieved.
- the inverted microscope according to the present embodiment can be replaced with an upright microscope as shown in FIG. 7, the same components as those in FIG. 5 are denoted by the same reference numerals.
- the optical members from the objective lens 31 to the imaging device 37 are disposed above the sealed container 20, and the transmission illumination device 38 is disposed below the sample chamber 10.
- the lens barrel of the objective lens 31 is inserted into the upper plate of the sealed container 20 via a seal member.
- the sample chamber 10 is moved in the X and Y directions by the XY movement stage 22.
- the light source of the transmission illumination device 38 is turned off.
- the biological sample S can be directly observed without passing through the culture vessel or the bottom plate of the well plate.
- the configuration of the environment preservation device is such that the temperature of the first chamber 110, the humidity, and the introduction part of the pipe in which the concentration of carbon dioxide is controlled are fixed parts (the second chamber one 20). By being provided on the surface (upper plate 21)) close to the first chamber, the environment in the first chamber 110 is controlled. In order to observe a microscope or an analyzer at the same temperature as the sample in the first chamber 110, it is desirable to set the second chamber 120 to the same temperature as the first chamber 110. In the above embodiment, the second chamber 1 does not necessarily need to be sealed as long as the control of the force and temperature illustrated as the closed container 20 is achieved. On the other hand, in order to prevent the controlled gas from leaking around the second chamber 120, it is preferable that the second chamber 120 has a sealed structure.
Abstract
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Priority Applications (2)
Application Number | Priority Date | Filing Date | Title |
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JP2005514171A JP4111223B2 (ja) | 2003-09-26 | 2004-09-16 | 環境保持装置および環境制御型分析装置 |
US11/388,201 US8066962B2 (en) | 2003-09-26 | 2006-03-24 | Environment holding apparatus and environment control type analyzer |
Applications Claiming Priority (2)
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JP2003335630 | 2003-09-26 | ||
JP2003-335630 | 2003-09-26 |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
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US11/388,201 Continuation US8066962B2 (en) | 2003-09-26 | 2006-03-24 | Environment holding apparatus and environment control type analyzer |
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WO2005031322A1 true WO2005031322A1 (ja) | 2005-04-07 |
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PCT/JP2004/013539 WO2005031322A1 (ja) | 2003-09-26 | 2004-09-16 | 環境保持装置および環境制御型分析装置 |
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US (1) | US8066962B2 (ja) |
JP (1) | JP4111223B2 (ja) |
WO (1) | WO2005031322A1 (ja) |
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JP2013076688A (ja) * | 2011-09-29 | 2013-04-25 | Samsung Electro-Mechanics Co Ltd | はんだボール検査装置 |
JP2016212115A (ja) * | 2010-09-08 | 2016-12-15 | テカン・トレーディング・アクチェンゲゼルシャフトTECAN Trading AG | 制御されたガス雰囲気を備えたマイクロプレートリーダーおよび対応する方法 |
JP2017181516A (ja) * | 2010-09-08 | 2017-10-05 | テカン・トレーディング・アクチェンゲゼルシャフトTECAN Trading AG | 制御されたガス雰囲気を備えたマイクロプレートリーダーおよび対応する方法 |
JP2021524911A (ja) * | 2018-05-25 | 2021-09-16 | ファイブ プライム セラピューティクス, インコーポレイテッド | 組織の特性評価とスクリーニングのために改善したサイトメトリー |
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KR100885182B1 (ko) * | 2006-12-20 | 2009-02-23 | 삼성전자주식회사 | 에미션 분석 장치 및 불량 분석 방법 |
JP5026849B2 (ja) * | 2007-04-20 | 2012-09-19 | 株式会社日立製作所 | 化学発光計測装置 |
US8561484B2 (en) | 2009-03-24 | 2013-10-22 | Perkinelmer Health Sciences, Inc. | Sorbent devices with longitudinal diffusion paths and methods of using them |
DE102010019776B4 (de) * | 2010-05-07 | 2015-07-02 | Thermo Electron Led Gmbh | Klimaschrank mit mehreren Ein- und Ausgabestationen |
JP5943375B2 (ja) | 2012-01-13 | 2016-07-05 | 矢崎総業株式会社 | 電気接続箱 |
JP6471853B2 (ja) * | 2015-01-26 | 2019-02-20 | 澁谷工業株式会社 | インキュベータ |
EP3500839B1 (en) * | 2016-08-16 | 2023-06-07 | Reacnostics GmbH | Application profile reactor for operando measurements |
DE102021114565B3 (de) | 2021-06-07 | 2022-09-08 | Leica Microsystems Cms Gmbh | Mikroskop und Verfahren zum Betreiben eines Mikroskops |
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Also Published As
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JPWO2005031322A1 (ja) | 2007-11-15 |
US8066962B2 (en) | 2011-11-29 |
US20060245976A1 (en) | 2006-11-02 |
JP4111223B2 (ja) | 2008-07-02 |
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