WO2005066683A1 - 対物レンズ、光分析装置、光分析装置の運転方法および顕微鏡 - Google Patents
対物レンズ、光分析装置、光分析装置の運転方法および顕微鏡 Download PDFInfo
- Publication number
- WO2005066683A1 WO2005066683A1 PCT/JP2005/000043 JP2005000043W WO2005066683A1 WO 2005066683 A1 WO2005066683 A1 WO 2005066683A1 JP 2005000043 W JP2005000043 W JP 2005000043W WO 2005066683 A1 WO2005066683 A1 WO 2005066683A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- lens
- objective lens
- tip
- container
- plate
- Prior art date
Links
Classifications
-
- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/0004—Microscopes specially adapted for specific applications
- G02B21/0088—Inverse microscopes
-
- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/33—Immersion oils, or microscope systems or objectives for use with immersion fluids
Definitions
- the present invention relates to an objective lens, a microscope and an optical analyzer using the objective lens, and an operation method thereof.
- the distance between the sample and the objective lens becomes a working distance.
- the lens surface or the lens frame of the objective lens comes in contact with the sample or the cover glass, the objective lens or the sample may be damaged.
- the distance between the objective lens and the specimen is set sufficiently in advance so as not to damage the objective lens and the specimen.
- the objective lens may be close to the sample, the cover glass, or the plate at a very small distance.
- the objective lens surface or lens frame force Sample, cover glass, plate Impact, and may damage the objective lens, sample, cover glass, and plate.
- the following confocal scanning optical microscope has been disclosed as a method for preventing damage to a sample or an objective lens due to the lens surface or lens frame of the objective lens coming into contact with the sample ( JP-A-8-190053).
- Sample observation range or measurement range To determine the enclosure, allow movement only in the direction where the objective lens and the sample approach first. Next, if the focal position of the objective lens is moved to the uppermost surface of the sample, then the upper limit is used as the upper limit. Be able to find the position that comes to the lowest surface of the upper surface of the sample. If the amount of movement exceeds the operating distance of the objective lens, stop the movement.
- a tactile sensor that detects contact of the sample with the objective lens
- a microscope system that moves the stage in a direction to move the sample away from the objective lens by a detection signal of the tactile sensor is provided. It is disclosed (see JP-A-2000-199858).
- the inner cylinder holding the tip lens of the objective lens is urged upward by a coil spring, and when the tip of the objective lens hits, for example, a container, the buffering action of the coil spring acts, and A microscope objective lens that prevents damage to the lens has also been disclosed (see JP-A-10-123426).
- Japanese Patent Application Laid-Open No. H8-190053 it is necessary to determine the observation range or measurement range of the sample for each observation site or measurement site of the sample. However, it takes a long time to do so. Also, Japanese Patent Application Laid-Open No. Hei 8-190053 describes that observation or measurement is performed while an objective lens is in contact with a sample, a cover glass, and a plate.
- Japanese Patent Application Laid-Open No. 2000-199858 each time the stage is moved in a direction away from the objective lens by the detection signal of the tactile sensor, the stage is moved or focused. It is necessary to make adjustments, which requires a lot of work time. Also, Japanese Patent Application Laid-Open No. 2000-199858 describes that observation or measurement is performed while an objective lens is in contact with a sample, a power bar glass, and a plate.
- the present invention relates to an objective lens, a microscope, an optical analyzer, and an optical analyzer that can reliably prevent damage to an objective lens, an object to be measured, a plate on which the object to be measured is loaded, or a container that houses the object to be measured. Provide driving method.
- the invention according to a first aspect of the present invention is an objective lens having a front lens and a lens frame supporting the front lens, wherein the lens frame is biased in a direction of a center axis of the front lens.
- the lens frame has an elastic member for supporting, and a front end of the lens frame has a convex portion protruding outside in the central axis direction from a front end surface of the front lens.
- the elastic member when measuring an object to be measured using the objective lens of the present invention, is formed of the object to be measured, a plate on which the object to be measured is loaded, or a container that holds the object to be measured.
- the lens frame is pressed against the surface facing the objective lens. For this reason, even if the object, plate or container moves in a plane perpendicular to the center axis of the tip lens and the position of the contact point in the center axis direction changes, the tip of the lens frame and the object Since the lens or the container is always in contact, measurement can be performed without collision between the lens frame and the object to be measured.
- the elastic member absorbs vibration at the time of contact between the object, the plate or the container and the objective lens, and absorbs a change in the position of the contact point in the central axis direction, so that vibration of the object to be measured can be reduced.
- the tip of the lens frame keeps the distance between the tip lens and the lens to be measured, the plate or the container, and the tip lens almost constant. Prevent contact between lens and object, plate or container.
- the distance from the tip lens to the object, plate, or container is almost constant before and after the object, plate, or container moves, so measurement can be performed without re-focusing at a new measurement point after movement. , A series of measurements can be performed quickly.
- D Working distance
- the distance from the position where the front end of the lens frame is the farthest away from the front end surface force of the front end lens in the direction of the central axis, that is, the convex portion, and the position where the front end lens is supported are zero.
- a self-lubricating resin member is used at the tip of the lens frame.
- the tip of the lens frame is less likely to be damaged when the tip of the lens frame comes into contact with the object, plate or container.
- the friction between the tip of the lens frame and the measurement object, plate or container is small, so that vibration of the measurement object, plate or container can be prevented.
- the stability of the measurement object can be maintained.
- the tip of the lens frame may be coated with fluorine resin.
- the front end of the lens frame has a convex portion protruding outside the front end surface of the front lens in the direction of the center axis of the front lens. That is, when the objective lens is set so that the front lens faces upward, the front end of the lens frame is at a position higher than the front end surface of the front lens. Then, the object to be measured, the plate on which the object to be measured is loaded, or the container accommodating the object to be measured is brought into contact with the tip of the lens frame. Are relatively moved in a plane perpendicular to the central axis of the.
- the tip of the lens frame becomes a spacer between the tip lens and the object to be measured, the plate or the container, and determines the distance from the contacted object to be measured, the plate or the container to the tip lens. Keep it almost constant to prevent contact between the tip lens and the object, plate or container.
- the plate, or the container moves while the tip of the lens frame is in contact with the object, plate, or container, measurement is performed without collision between the lens frame and the object, the plate, or the container. be able to.
- moving the measurement object, plate or container Before and after the measurement the distance from the front lens to the object to be measured, the plate or the container is almost constant, so that measurement can be performed without refocusing at a new measurement point after moving, and a series of measurements can be performed. Can be done quickly.
- D Working distance
- the distance from the tip lens to the object, plate, or container is constant. Measurement can be performed.
- the objective lens moves along the central axis direction and comes into contact with the object to be measured, the plate or the container.
- the movement of the objective lens suppresses the vibration of the object to be measured, makes accurate contact, and allows the objective lens to move quickly.
- the measurement object, plate, or container moves in a plane perpendicular to the center axis of the objective lens, so that the objective lens and the optical system before and after the objective lens during measurement, such as a light source, an optical path, a mirror, and a detector. Since the position is fixed, measurement can be performed with high accuracy.
- the focal position when determining the focal position of the objective lens, the focal position is determined within a range where the measurement target exists. Based on the maximum displacement of the contact surface between the container and the front end of the lens frame in the direction of the central axis, it is determined that the distance from the boundary of the measurement object in the direction of the central axis, for example, the upper and lower limits, is greater than the maximum displacement. is there. According to the sixth aspect, even if the contact surface is displaced in the direction of the central axis, the focal point exists in the measurement target, and once the focal position is determined, even if the measurement target, plate, or container is moved, The measurement can be started immediately without focusing each time.
- the distal end of the lens frame has a convex portion protruding outward from the distal end surface of the distal lens in the direction of the center axis of the distal lens. That is, when the objective lens is set so that the front lens faces upward, the front end of the lens frame is located higher than the front end surface of the front lens. Further, the contact means makes the tip of the lens frame come into contact with the object to be measured, the plate on which the object to be measured is loaded, or the container accommodating the object to be measured. Container and objective lens Are relatively moved within a plane perpendicular to the center axis of the objective lens.
- the tip of the lens frame forms a spacer between the tip lens and the object to be measured, the plate or the container, and determines the distance from the contacting object, plate or container to the tip lens. Keep it almost constant to prevent contact between the tip lens and the object, plate or container.
- the plate, or the container moves while the tip of the lens frame is in contact with the object, plate, or container, measurement is performed without collision between the lens frame and the object, the plate, or the container. be able to.
- the plate or the container is almost constant, so that measurement can be performed without refocusing at a new measurement point after the movement. A series of measurements can be performed quickly.
- the distance WD working distance from the front lens to the focal point is constant.
- the distance from the tip lens to the object, plate, or container is constant, even if the object, plate, or container moves, the focal point is in the sample, so that the measurement can be performed reliably.
- the distal lens and the lens frame supporting the distal lens are supported by the elastic member by applying a biasing force in the direction of the center axis of the lens frame.
- the elastic member presses the lens frame against the surface of the object to be measured, the plate, or the container facing the objective lens, the contact portion moves in a plane perpendicular to the center axis of the tip lens and the contact member moves. Even if the position in the direction of the central axis changes, the tip of the lens frame is always in contact with the object to be measured, the plate or the container.
- the elastic member absorbs vibration at the time of contact between the object to be measured, the plate or the container, and the objective lens, and absorbs a change in the position of the contact point in the central axis direction, thereby reducing vibration of the object to be measured.
- the objective lens moving device moves the objective lens in the direction of the central axis to bring the measurement object, plate or container into contact with the tip of the lens frame. According to the ninth surface, the movement for contact can be performed accurately and quickly by moving the objective lens.
- the objective lens frame is supported by the elastic member exerting a biasing force in the direction of the center axis of the objective lens frame. According to the tenth aspect, the elastic member presses the objective lens against a surface of the measurement object, a plate on which the measurement object is loaded, or a container that houses the measurement object, facing the objective lens.
- the tip of the lens frame always contacts the object to be measured, the plate or the container. I have.
- the elastic member can absorb a change in the position of the contact point in the direction of the central axis, thereby reducing the vibration of the object to be measured.
- the stage supports the object to be measured, the plate or the container, and regulates movement in the direction of the central axis.
- the vibration of the measurement target can be reduced, and the stability of the measurement target can be maintained.
- the stage supports the object to be measured, the plate or the container, and is powered in a plane perpendicular to the direction of the central axis.
- the movement of the stage does not affect the optical system before and after the objective lens and the objective lens, for example, a light source, an optical path, a mirror, a detector, and the like.
- the movement of the objective lens in a plane perpendicular to the central axis is restricted.
- the positions of the optical systems before and after the objective lens for example, the light source, the optical path, the mirror, and the detector are fixed. Therefore, the measurement can be performed with higher accuracy.
- the tip lens and the lens frame supporting the tip lens are supported by the elastic member by applying a biasing force in the direction of the center axis of the lens frame. Further, when the front end of the lens frame is located outside the front end surface of the front lens in the direction of the center axis of the front lens, that is, when the objective lens is set so that the front lens is directed upward, the front end of the lens frame becomes the front lens.
- the objective lens moving device moves the objective lens in the direction of the central axis to bring the object to be measured, the plate or the container into contact with the tip of the lens frame, the stage supports the object to be measured, the plate or the container, and It moves in a plane perpendicular to the central axis while being restricted from moving in the axial direction.
- the elastic member pairs with the objective lens of the object to be measured, the plate or the container. Since the lens frame is pressed against the facing surface, even if the contact point moves in a plane perpendicular to the center axis of the tip lens and the position of the contact point in the center axis direction changes, the tip of the lens frame and the target Always in contact with plates or containers.
- the elastic member absorbs vibration at the time of contact between the object to be measured, the plate or container, and the objective lens, and absorbs changes in the position of the contact point in the central axis direction, thereby reducing vibration of the object to be measured. Can be.
- the tip of the lens frame acts as a spacer between the tip lens and the object to be measured, the plate or container, and keeps the distance between the object to be measured, the plate or the container, and the tip lens almost constant. Prevents contact with measurement object, plate or container.
- the measurement object, plate, or container moves while the tip of the lens frame is in contact with the measurement object, plate, or container.Measurements must be made without collision between the lens frame and the measurement object, plate, or container. Can be.
- the distance from the front lens to the measurement object, plate, or container is almost constant, so measurement can be performed at a new measurement point after movement without refocusing.
- D Working distance
- the distance from the tip lens to the object, plate, or container is constant. Measurement can be performed.
- the movement for contact can be performed accurately and quickly by moving the objective lens.
- the movement of the stage does not affect the objective lens and the optical system before and after the objective lens, for example, a light source, an optical path, a mirror, a detector, etc., so that the measurement can be performed with high accuracy. Since the movement of the object, the plate or the container in the direction of the central axis is restricted, the vibration of the object to be measured can be reduced, and the stability of the object to be measured can be maintained.
- the distal end of the lens frame has a projection protruding outward in the central axis direction from the distal end surface of the distal lens, and the objective lens is positioned so that the distal lens faces upward.
- the top of the lens frame is higher than the top of the front lens.
- the objective lens moving device moves the objective lens in the direction of the central axis, and brings the measurement object, plate or container into contact with the tip of the lens frame.
- the elastic member supports the objective lens by applying a biasing force in the direction of the central axis, the stage supports the object to be measured, the plate or the container, and the movement in the direction of the central axis is restricted while being perpendicular to the direction of the central axis. To move in a perfect plane.
- the tip of the lens frame measures the distance between the tip lens and the object to be measured, the force of the object to be measured in contact with the plate or the container, and the plate or container. Keep it almost constant to prevent contact between the tip lens and the object, plate or container.
- the elastic member presses the lens frame against the surface of the measurement object, plate or container facing the objective lens. Therefore, even if the contact point moves in a plane perpendicular to the center axis of the tip lens and the position of the contact point in the direction of the center axis changes, the tip of the lens frame is always in contact with the measurement object, plate, or container. .
- the elastic member absorbs vibration at the time of contact between the object to be measured, the plate or the container, and the objective lens, and absorbs a change in the position of the contact point in the central axis direction, thereby reducing vibration of the object to be measured.
- the plate, or the container moves while the tip of the lens frame is in contact with the object, plate, or container, measurement is performed without collision between the lens frame and the object, the plate, or the container. be able to. Also, before and after the movement, the distance from the tip lens to the object to be measured, the plate or the container is almost constant, so that measurement can be performed at a new measurement point after the movement without refocusing. In addition, a series of measurements can be performed quickly.
- the distance WD working distance from the front lens to the focal point is constant.
- the distance from the tip lens to the object, plate, or container is constant, even if the object, plate, or container moves, the focal point is in the sample, so that the measurement can be performed reliably.
- the movement for contact can be performed accurately and quickly by moving the objective lens.
- the movement of the stage is controlled by the objective lens and the optical system before and after the objective lens. It does not affect the system, for example, light source, light path, mirror, detector, etc., so that the measurement can be performed with high accuracy.Also, since the movement of the measurement object, plate or container in the central axis direction is regulated, Vibration can be reduced, and the stability of the measurement object can be maintained.
- the front end of the lens frame has a convex portion protruding outward from the front end surface of the front end lens in the direction of the central axis of the front end lens, and the front end lens faces upward.
- the tip of the lens frame is located higher than the tip surface of the tip lens, and the adjustment device contacts the observation target, plate or container with the tip of the lens frame.
- the front end of the lens frame forms a spacer between the front lens and the object to be measured, the plate or the container, and the force of the object to be measured, the plate or the container that has come into contact with the front lens is almost equal to the distance to the front lens. Keep the tip constant to prevent contact between the tip lens and the object, plate or container.
- the adjustment device will adjust the front end of the lens frame and the observation object. , Always in contact with plates or containers. Therefore, when the observation target, the plate, or the container is moved, observation can be performed without collision between the lens frame and the observation target.
- the distance WD working distance from the front lens to the focal point is reduced. Since the distance from the tip lens to the observation target, plate, or container is constant, even if the observation target, plate, or container moves, the focal point is in the sample, so that observation can be reliably performed.
- FIG. 1 is a diagram showing a schematic configuration of a fluorescence analyzer according to a first embodiment of the present invention.
- FIG. 2 is a cross-sectional view in the lens axis direction of an immersion objective lens used in the first embodiment.
- FIG. 3 is a diagram showing a tip of a lens frame of a liquid immersion objective lens used in the first embodiment.
- FIG. 4 is a diagram showing a schematic configuration of a control unit used in the first embodiment.
- FIG. 5 is a flowchart showing an operation procedure of the fluorescence analyzer according to the first embodiment.
- FIG. 6 is a view for explaining the relationship between the position of the focal point of the immersion objective lens according to the first embodiment and the displacement of the bottom surface of the microplate.
- FIG. 7A is a diagram illustrating the relationship between the position of the focal point of the liquid immersion objective lens according to the first embodiment and the displacement of the bottom surface of the microplate.
- FIG. 7B is a view for explaining the relationship between the position of the focal point of the immersion objective lens according to the first embodiment and the displacement of the bottom surface of the microplate.
- FIG. 8 is a diagram showing a schematic configuration of an objective lens moving device used in a second embodiment of the present invention.
- FIG. 1 is a diagram showing a schematic configuration of a fluorescence analyzer which is an optical analyzer to which the present invention is applied.
- the fluorescence analyzer main body 10 includes a light source 5, an inverted fluorescence microscope 1 using confocal optics, and an electric signal converter that acquires fluorescence emitted from a fluorescently labeled sample and converts it into an electrical signal.
- the photoelectric conversion unit 2 is a means
- the data processing unit 3 is a data processing unit that determines the characteristics of the sample based on the measurement data obtained by the photoelectric conversion unit 2, and the measurement data is obtained by the data processing unit 3.
- a display device 4 for displaying various characteristics of the sample based on the information.
- the fluorescence analyzer main body 10 further includes a control unit 6 (see FIG. 3 described later) for controlling the above-described units.
- the light source 5 includes, for example, a laser light generator.
- the photoelectric conversion unit 2 includes a photomultiplier and an avalanche photodiode.
- the inverted fluorescence microscope 1 includes a stage 13 that supports a microplate 12 on which a measurement target is mounted.
- the stage 13 has a moving device 21 and a drive control device 22.
- the drive unit 15 is provided.
- the moving device 21 moves the stage 13 in the XY directions so that the microplate 12 moves in a plane perpendicular to the optical axis 28. That is, the movement of the stage 13 in the direction of the optical axis 28 is restricted, and only movement in a plane perpendicular to the optical axis 28 is allowed.
- the drive control device 22 controls the drive of the moving device 21 based on a command from the control unit 6.
- the microplate 12 is formed with a plurality of recesses for accommodating a sample to be measured and through which light from the light source 5 is transmitted.
- an immersion objective lens 11 is arranged close to the microplate 12.
- the immersion objective lens 11 is provided with an objective lens moving device 23.
- the immersion objective lens 11 can be driven in the direction of the optical axis 28 by the drive control of the objective lens moving device 23 by the drive control device 301 based on a command from the control section 6.
- a liquid supply recovery device 14 for supplying a liquid is provided around the immersion objective lens 11 and the microplate 12.
- the liquid supply / recovery device 14 is attached to a supply nozzle 19 for supplying a liquid 24 between the bottom surface 18 of the microplate 12 and the immersion objective lens 11, and attached to the outer periphery of the immersion objective lens 11.
- the liquid receiving tray 25 that prevents the supplied liquid 24 from flowing into the device below and in the details of the immersion objective lens 11, and the liquid that collects the liquid from the liquid receiving tray 25 by adjusting the amount of the supplied liquid 24. It has an amount adjusting device 20 and the like.
- a dichroic mirror 291 is arranged on the optical axis 28 of the immersion objective lens 11.
- the dichroic mirror 291 has a property of reflecting short-wavelength light and transmitting long-wavelength light, and has a wavelength of excitation light incident from the light source 5 via the illumination optical path 16.
- the light is reflected by the immersion objective lens 11, transmits the light of the fluorescence wavelength incident from the immersion objective lens 11, and is incident on the photoelectric conversion section 2 via the light detection optical path 17.
- the stage 13 supporting the microplate 12 above the immersion objective lens 11 moves the microplate 12 in a plane perpendicular to the optical axis 28.
- the vibration of the sample in the microplate 12 can be reduced, and the state of the sample can be kept stable.
- the microplate 12 moves only in a plane perpendicular to the optical axis 28,
- the positional relationship between the immersion objective lens 11 and the optical system before and after the immersion objective lens 11, for example, the light source 5, the illumination optical path 16, the light detection optical path 17, and the photoelectric conversion unit 2 can be fixed. . Therefore, the optical measurement accuracy can be stably maintained.
- FIG. 2 shows a sectional view of the immersion objective lens 11 in the lens axis direction.
- the immersion objective lens 11 has a cylindrical lens frame 32.
- a front end lens 31 and a lens group 30 are accommodated in the hollow portion of the lens frame 32 along the optical axis 28.
- the lens frame 32 has a lens frame tip 36 that protrudes outward in the direction of the optical axis 28 (the center axis of the tip lens 31) from the tip face 31a of the tip lens 31.
- the front end 36 of the lens frame is positioned higher than the front end surface 31 a of the front lens 31.
- the front end 36 of the lens frame is located farthest from the front end surface 31a of the front end lens 31 in the direction of the center axis and has the maximum distance (convex portion 36a), and the position where the front end lens 31 is supported. These distances are equal to or greater than zero and smaller than the maximum distance! / ⁇ (recess 36b) is formed.
- the concave portion 36b is formed so that when the liquid 24 is supplied onto the immersion objective lens 11, the liquid 24 spreads inside and outside the lens frame front end 36.
- a resin having self-lubricating properties is used as the material of the lens frame tip 36.
- a soft material such as polyacetal is preferable as the material of the lens frame tip 36.
- the front end 36 of the lens frame contacts the bottom surface 18 of the microplate 12
- the bottom surface 18 is scratched.
- the microplate 12 moves, the friction between the front end 36 and the bottom surface 18 of the lens frame is small, so that the vibration to the microplate 12 can be prevented, and the stability of the sample can be maintained. Can be.
- a series of measurements can be performed quickly.
- the end of the lens frame 32 opposite to the front end 36 of the lens frame is movably inserted into the hollow portion of the outer cylinder 34.
- the outer cylinder 34 is fixed to an objective lens mount 35 of the objective lens moving device 23.
- an elastic member 33 is disposed between the lens frame 32 and the objective lens mount 35. The elastic member 33 exerts a biasing force such that the lens frame 32 is parallel to the optical axis 28 (in the direction of the center axis of the front lens 31) and is directed toward the microphone opening plate 12 (upward on the paper) with respect to the objective lens mount 35.
- the elastic member 33 Has a role of absorbing vibration generated when the lens frame tip 36 of the lens frame 32 contacts the bottom surface 18 of the microplate 12, and a role of absorbing vibration while maintaining these contacts.
- a force using a metal spring, an air spring, or the like may be used.
- a material having a restoring force or a repulsive force of a magnet may be used.
- a liquid receiving plate 25 used in the liquid supply and recovery device 14 described above is provided around the lens frame 32.
- the objective lens moving device 23 has a cylindrical hollow portion 23a formed along the optical axis 28.
- An objective lens mount 35 is provided movably along the optical axis 28 in the hollow portion 23a.
- the objective lens moving device 23 includes a feed mechanism 38 for moving the objective lens mount 35 in the direction of the optical axis 28, and a guide 39 for maintaining the objective lens mount 35 in the hollow portion 23a in the horizontal direction.
- the microplate 12 moves in the X-Y direction, that is, in the horizontal direction (left / right / front of the paper in FIG. 1), and the position of the bottom surface 18 of the microplate 12 with respect to the optical axis 28. Even if changes, the front end 36 of the lens frame always comes into contact with the bottom surface 18 stably.
- the immersion objective lens 11 can be moved quickly, and the immersion objective lens 11 can be moved quickly.
- FIG. 4 shows details of the control unit 6.
- control unit 6 has a central control unit (CPU) 29, and the central control unit 29 controls the light source 5, the illumination optical path 16, and the optical system of the light detection optical path 17 by a light source′optical A system controller 7, a stage controller 8 for controlling the movement of the stage 13, a liquid supply controller 9 for controlling the liquid supply / recovery device 14, and an objective lens for controlling the movement and focusing of the liquid immersion objective lens 11.
- Control unit 281 is connected.
- the central controller 29 controls the light source ⁇ ⁇ optical system controller 7, stage controller 8, liquid supply controller 9, and objective lens controller in accordance with a predetermined operation procedure of the fluorescence analyzer 10 (see FIG. 5). 28 is given a predetermined instruction.
- the microplate 12 is set on the stage 13 (Step 501). Then, the central control unit 29 of the control unit 6 moves the stage 13 to a predetermined position, for example, a position where a specific well of the microplate 12 is on the optical axis 28 of the immersion objective lens 11. To the stage control unit 8.
- the stage control unit 8 sends a stage movement command to the drive control device 22, and the drive control device 22 gives the stage movement amount to the movement device 21. Accordingly, the moving device 21 moves the stage 13 by an amount based on the instruction of the stage control unit 8 (step 502).
- the central control unit 29 instructs the liquid supply control unit 9 to supply the liquid onto the tip lens 31 of the immersion objective lens 11.
- the liquid supply control unit 9 drives the liquid amount adjusting device 20 according to an instruction from the central control unit 29.
- the liquid amount adjusting device 20 supplies a predetermined amount of the liquid 24 from the supply nozzle 19 onto the distal end surface 31a of the distal lens 31 surrounded by the lens frame distal end 36 of the liquid immersion objective lens 11, and supplies the liquid bulb. Make it (Step 503). At this time, the liquid 24 overflowing from the front end 36 of the lens frame around the front end lens 31 is collected in the liquid receiving tray 25 and then collected by the liquid amount adjusting device 20.
- the central controller 29 instructs the objective lens controller 28 to move and focus the immersion objective lens 11, and further supplies light to the light source / optical system controller 7. Command.
- the objective lens control unit 28 sends an instruction for moving the immersion objective lens 11 to the objective lens moving device 23.
- the objective lens moving device 23 drives the feed mechanism 38 to move the objective lens mount 35 in the direction of the microplate 12 (upward on the paper) (step 504).
- the biasing force of the elastic member 33 is applied to the bottom surface 18 of the microplate 12 from the front end 36 of the lens frame at a position where the front end 36 of the lens frame contacts the bottom surface 18 of the microplate 12.
- the distortion of the microplate 12 and the thickness of the bottom are uneven.
- the position of the bottom surface 18 of the microplate 12 in the height direction may change even if the bottom surface 18 of the microplate 12 is undulated even if the thickness of the microplate 12 is uniform. Measurement can be performed without collision between the frame 32 and the bottom surface 18 of the microplate 12.
- the elastic force of the elastic member 33 can absorb the change in the position of the bottom surface 18 of the microplate 12 and reduce the vibration of the sample 231 described later in the microplate 12, so that the sample 231 can be stably maintained. can do.
- the objective lens control unit 28 performs focusing of the immersion objective lens 11 (step 505). ).
- the position of the focal point 27 is determined based on a predetermined maximum displacement At of the bottom surface 18 of the microplate 12 and the bottom surface 26 a of the well 26 and the liquid surface 231 a of the sample 231. Between the bottom 26a and the liquid surface 231a of the well 26 and below the At and a sufficient depth of the well 26 is determined.
- the central control unit 29 instructs the data processing unit 3 to measure the fluorescence and start the data processing.
- the data processing unit 3 performs measurement and data processing according to the instruction of the central control unit 29 (Step 506).
- the photoelectric signal converter 2 acquires the fluorescence emitted from the fluorescently labeled sample 231 and converts it into an electric signal to obtain measurement data.
- the data processing unit 3 obtains the characteristics of the sample 231 based on the obtained measurement data.
- the central control unit 29 instructs the stage control unit 8 to move the stage 13 so that the next measurement target jewel 26 comes above the immersion objective lens 11.
- the stage control section 8 sends a stage movement command to the drive control device 22.
- the drive control device 22 gives the moving amount to the moving device 21.
- the moving device 21 is The plate 12 is moved horizontally (step 507).
- the fluorescence measurement and the movement of the stage 13 are repeated in steps 506 and 507 until the measurement in the well 26 scheduled as the measurement object is completed (NO in step 508).
- the central control unit 29 instructs the objective lens control unit 28 to move the immersion objective lens 11 away from the microplate 12.
- the objective lens controller 28 commands the objective lens moving device 23 to move the immersion objective lens 11.
- the objective lens moving device 23 drives the feed mechanism 38 to move the objective lens mounting base 35 away from the microplate 12 (downward on the paper) (step 509). Thereafter, the process is terminated by replacing the microplate 12 (step 510).
- the distance d from the tip surface 31a of the tip lens 31 to the focal point 27 is the distance g from the tip surface 31a to the bottom surface 18 of the microplate 12, and the bottom g of the microplate 12 The sum of the distance h from the bottom surface 26a of the hole 26 to the focal point 27.
- the focal length of the immersion objective lens 11 is fixed, the distance h from the bottom surface 26a of the well 26 to the focal point 27 depends on the distance g from the tip surface 31a to the bottom surface 18 of the microplate 12. Since the bottom of the microplate 12 is a solid medium, the distance h does not depend on the thickness t of the bottom of the microplate 12 AtZ2. As a result, as the distance g increases when the immersion objective lens 11 is further away from the microplate 12, the distance h decreases, and finally the inner force of the well 26 is deviated.
- the displacement g of the distance g from the front end surface 31a of the front lens 31 to the bottom surface 18 is changed. Is less than or equal to the displacement of the bottom surface 18 (ie, very small). That is, the distance g is almost constant and Therefore, the distance h is almost constant.
- the position of the focal point 27 of the immersion objective lens 11 is determined between the bottom surface 26a and the liquid surface 231a of the sample 231.
- the maximum displacement ⁇ beam due to non-uniformity of the bottom surface 18 is higher than the beam and the maximum displacement ⁇ beam is lower than the liquid surface 231a, even if the bottom surface 18 is displaced, the focal point 27 exists in the sample 231. I do. Therefore, after the focal position is determined by the operation of step 504, even if the measurement target well 26 is moved, the lens frame tip 36 is in contact with the bottom surface 18 of the microplate 12, so that The measurement can be started immediately without having to focus on the degree. The same applies to the case where the entire microplate 12 is distorted. Since the front end 36 of the lens frame is always in contact with the bottom surface 18 of the microplate 12, even if the well 26 is moved, the focal point 27 exists in the sample, The measurement can be started immediately.
- the front end lens 31 and the lens frame 32 supporting the front end lens 31 are supported by the elastic member 33 exerting a biasing force in the direction of the center axis of the lens frame 32.
- the distal end of the lens frame 32 is protruded from the distal end face 31a of the distal lens 31 to the outside in the direction of the central axis of the distal lens 31, and the objective lens is set so that the distal lens 31 faces upward, the lens The tip of the frame 32 is higher than the tip surface of the tip lens 31 and at a position!
- the objective lens moving device 23 moves the immersion objective lens 11 in the direction of the central axis, and makes contact with the bottom surface 18 of the microphone port plate 12.
- the stage 13 supports the microplate 12 and moves only in a plane perpendicular to the central axis direction while being restricted from moving in the central axis direction.
- the lens frame 32 can be pressed against the surface of the microplate 12 facing the immersion objective lens 11 by the elastic member 33. Even if the microplate 12 is distorted or the thickness of the bottom is not uniform, or the thickness of the bottom plate is undulating even if the thickness is uniform, the displacement of the position of the bottom surface 18 causes the lens frame tip 36 to be displaced. The state in which the microplate 12 is always in contact with the bottom surface 18 can be maintained. Further, by moving the microplate 12 in the horizontal direction, measurement for fluorescence analysis can be performed without the microplate 12 colliding with the immersion objective lens 11.
- the elastic member 33 absorbs vibration at the time of contact between the microplate 12 and the immersion objective lens 11 and absorbs changes in the position of the contact point in the direction of the central axis. Vibration can be reduced.
- the tip of the lens frame 32 plays a role of a spacer between the tip lens 31 and the microplate 12, the distance from the contacting microplate 12 to the tip lens 31 can be kept almost constant. Thus, contact between the tip lens 31 and the microplate 12 can be reliably avoided.
- the microphone opening plate 12 is moved in the horizontal direction in a state where the lens frame tip 36 of the lens frame 32 and the microplate 12 are in contact with each other, the lens frame 32 and the microplate 12 do not collide with each other. A measurement can be made. Furthermore, before and after the movement of the microplate 12, the distance from the tip lens 31 to the microplate 12 can be made substantially constant, so that measurement can be performed at a new measurement point after the movement without refocusing. Therefore, a series of measurements can be performed quickly.
- the distance WD working distance from the tip lens 31 to the focal point.
- the distance from the tip lens 31 to the microplate 12 can be made constant. Therefore, even if the microplate 12 moves, the focus is in the sample, so that the measurement can be reliably performed.
- the immersion objective lens 11 side Is moved in the direction of the microplate 21 along the direction of the optical axis 28 (the center axis of the tip lens 31), so that the movement for the contact during this time can be quickly performed with high accuracy.
- the movement of the stage 13 at the time of measurement can be performed without affecting the immersion objective lens 11 and the optical system before and after the immersion objective lens 11, such as a light source, an optical path, a mirror, and a detector. it can. Therefore, measurement can be performed with high accuracy.
- the vibration of the object to be measured can be reduced, and the stability of the object to be measured can be maintained.
- FIG. 8 is a diagram showing another example of the objective lens moving device 23, and the same parts as those in FIG. 2 are denoted by the same reference numerals.
- the lens frame 32 of the immersion objective lens 11 is fixed to the outer cylinder 34.
- a drive base 40 is provided in the objective lens moving device 23.
- the drive base 40 has a cylindrical hollow portion 40a along the optical axis 28.
- An objective lens mount 35 is provided movably along the optical axis 28 in the hollow portion 40a.
- the drive base 40 is provided with a guide 41 for maintaining the objective lens mounting base 35 in the drive base 40 in a horizontal direction.
- the movement of the objective lens mount 35 to which the immersion objective lens 11 is mounted in a plane perpendicular to the optical axis 28 is restricted, and the optical axis 28 (the tip lens 3 1 Only the movement in the direction of the center axis) is allowed.
- a through hole 40b is formed in the side surface of the drive base 40.
- An operation arm 35a is provided on a side surface of the objective lens mount 35. The operating arm 35a penetrates the through hole 40b on the side surface of the drive base 40 and protrudes to the outside.
- An elastic member 42 is provided between the distal end of the operation arm 35a and the protrusion 40c on the side surface of the drive stand 40.
- the elastic member 42 is biased such that the objective lens mounting base 35 is parallel to the optical axis 28 (in the direction of the center axis of the front end lens 31) and faces the microplate 12 (upward on the paper) with respect to the drive base 40. Acts on bias.
- the elastic member 42 serves to absorb vibration generated when the front end 36 of the lens frame 32 contacts the bottom surface 18 of the microplate 12, and absorbs vibration while maintaining these contacts. Have a role.
- a force using a metal spring, an air spring, or the like is used for the elastic member 42.
- a material having a restoring force or a magnet attracting force is used instead of the elastic member 42. You may.
- the objective lens moving device 23 includes a feed mechanism 38 for moving the drive base 40 in the direction of the optical axis 28, and a guide 39 for maintaining the horizontal position of the drive base 40. This restricts the movement of the objective lens mount 35 on which the immersion objective lens 11 is mounted in a plane perpendicular to the optical axis 28, and allows only movement in the direction of the optical axis 28 (the central axis of the front lens 31). It has been. [0086] Others are the same as those in FIG.
- the drive table 40 is moved in the direction of the microplate 12 (upward on the paper) by the feed mechanism 38, and the lens is moved by the biasing force of the elastic member 42. Set so that the frame tip 36 is pressed against the bottom surface 18 of the microplate 12. As a result, the front end 36 of the lens frame comes into contact with the bottom surface 18 of the microplate 12 at all times, and the operation is performed in the same procedure as described in the first embodiment.
- the elastic member is built in the immersion objective lens 11, and in any case, by mounting the objective lens mounting base 35 to which the immersion objective lens 11 is mounted on the drive base 40, the elastic member 42 Accordingly, a pulling force can be applied to the drive base 40 so that the objective lens mount 35 is directed parallel to the optical axis 28 toward the microplate 12 (upward on the paper).
- the immersion objective lens 11 is moved to the microplate 12 side together with the drive stand 40 by the objective lens moving device 23 from this state, so that the front end 36 of the lens frame is always in contact with the bottom surface 18 of the microplate 12.
- the same operation and effect as those of the first embodiment can be obtained.
- a resin having self-lubricating properties is used as the material of the lens frame tip 36.
- the lens frame 32 is formed using the same material as the lens frame 32.
- the tip 36 may be coated with fluorine resin.
- a rolling member may be provided on the lens frame 32 so that the lens frame 32 rolls and slides while contacting the microplate 12 by a bearing or the like.
- the bottom surface 18 of the microplate 12 may be lubricated.
- an objective lens used in a force-dry state using the immersion objective lens 11 may be used as the objective lens.
- the case where a microscope is used has been described.
- the same operation and effect can be obtained also in the case of observation using a microplate reader.
- the microplate 12 is used in the first and second embodiments, the present invention can be applied to observation using slide glass instead of the microplate 12 as a container for storing the measurement target. is there.
- slide glass instead of the microplate 12 as a container for storing the measurement target.
- the present invention can be applied to observation using slide glass instead of the microplate 12 as a container for storing the measurement target. is there.
- slide glass by bringing the front end 36 of the lens frame into contact with the lower surface of the slide glass, when the slide glass is powered, the focus is maintained in the sample on the slide glass even if the lower surface of the slide glass is distorted. Therefore, the same function and effect can be obtained.
- an upright microscope instead of an inverted microscope When the lens is used, the focal point is maintained in the sample by bringing the front end 36 of the lens frame into contact with the cover glass, and the same operation and effect can be obtained.
- the present invention is not limited to the above-described embodiment, and can be variously modified in a range without changing the gist thereof in an implementation stage.
- an objective lens a microscope, an optical analyzer, and a method of operating an optical analyzer that can reliably prevent damage to an objective lens, a measurement object, a plate on which the measurement object is loaded, or a container that stores the measurement object.
Abstract
Description
Claims
Priority Applications (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP05703315A EP1703312A1 (en) | 2004-01-07 | 2005-01-05 | Objective, optical analyzer, method of driving optical analyzer, and microscope |
US11/482,273 US7274433B2 (en) | 2004-01-07 | 2006-07-07 | Objective, optical analyzer, method of driving optical analyzer, and microscope |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2004002107A JP2005195878A (ja) | 2004-01-07 | 2004-01-07 | 対物レンズ、光分析装置、光分析装置の運転方法および顕微鏡 |
JP2004-002107 | 2004-01-07 |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US11/482,273 Continuation US7274433B2 (en) | 2004-01-07 | 2006-07-07 | Objective, optical analyzer, method of driving optical analyzer, and microscope |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2005066683A1 true WO2005066683A1 (ja) | 2005-07-21 |
Family
ID=34747015
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/JP2005/000043 WO2005066683A1 (ja) | 2004-01-07 | 2005-01-05 | 対物レンズ、光分析装置、光分析装置の運転方法および顕微鏡 |
Country Status (4)
Country | Link |
---|---|
US (1) | US7274433B2 (ja) |
EP (1) | EP1703312A1 (ja) |
JP (1) | JP2005195878A (ja) |
WO (1) | WO2005066683A1 (ja) |
Families Citing this family (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP1717628B1 (en) * | 2004-02-16 | 2012-01-25 | Olympus Corporation | Immersion objective lens, retention mechanism for immersion medium and manufacturing method thereof |
TWM295766U (en) * | 2006-02-14 | 2006-08-11 | Quanta Comp Inc | Input pen storage |
JP5266855B2 (ja) * | 2008-04-21 | 2013-08-21 | 日本電気株式会社 | 光学式外観検査装置 |
US8687180B2 (en) * | 2012-06-07 | 2014-04-01 | Molecular Devices, Llc | System, method, and device for determining a focal position of an objective in a microscopy imaging system |
US9297692B2 (en) * | 2013-02-20 | 2016-03-29 | Applied Materials Israel, Ltd. | System and method for inspecting a sample using landing lens |
DE102013207712A1 (de) * | 2013-04-26 | 2014-10-30 | Hamilton Bonaduz Ag | Mikroskop mit mechanischem Autofokus und Verfahren zum Fokussieren auf ein zu mikroskopierendes Objekt |
US10948707B2 (en) * | 2019-02-05 | 2021-03-16 | Molecular Devices, Llc | Liquid immersion microscope objective assembly and related systems and methods |
JP7254422B2 (ja) * | 2019-04-09 | 2023-04-10 | 株式会社ミツトヨ | 表面形状測定システムおよび表面形状測定器を用いた表面形状測定方法 |
US11747605B2 (en) * | 2019-04-23 | 2023-09-05 | Molecular Devices, Llc | System and method for formation and detection of immersion fluid boluses on a microscope objective |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS59109310U (ja) * | 1983-01-13 | 1984-07-23 | 富士通株式会社 | 顕微鏡 |
JPS59147107U (ja) * | 1982-10-26 | 1984-10-01 | 株式会社富士電機総合研究所 | 先端レンズ損傷防止用保護具 |
JP2003029162A (ja) * | 2001-07-11 | 2003-01-29 | Nikon Corp | 顕微鏡用対物レンズおよび顕微鏡 |
JP2003185926A (ja) * | 2001-12-20 | 2003-07-03 | Nikon Corp | 安全装置付液浸対物レンズ |
Family Cites Families (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS59109310A (ja) | 1982-12-15 | 1984-06-25 | Royal Kogyo Kk | 成形方法及び成形機 |
JPS59147107A (ja) | 1983-02-10 | 1984-08-23 | 三井造船株式会社 | 2個のハニカム板の接続方法 |
JP3573508B2 (ja) | 1995-01-11 | 2004-10-06 | オリンパス株式会社 | 共焦点走査型光学顕微鏡 |
JPH10123426A (ja) | 1996-10-22 | 1998-05-15 | Nikon Corp | 顕微鏡用対物レンズ |
JP4071882B2 (ja) | 1999-01-05 | 2008-04-02 | オリンパス株式会社 | 顕微鏡システム |
US7109464B2 (en) * | 2001-07-06 | 2006-09-19 | Palantyr Research, Llc | Semiconductor imaging system and related methodology |
JP2003156668A (ja) * | 2001-11-20 | 2003-05-30 | Canon Inc | 光学ユニット、露光装置および光学機器 |
EP3352010A1 (en) * | 2003-04-10 | 2018-07-25 | Nikon Corporation | Run-off path to collect liquid for an immersion lithography apparatus |
US6954256B2 (en) * | 2003-08-29 | 2005-10-11 | Asml Netherlands B.V. | Gradient immersion lithography |
US7088422B2 (en) * | 2003-12-31 | 2006-08-08 | International Business Machines Corporation | Moving lens for immersion optical lithography |
JP4253592B2 (ja) * | 2004-01-06 | 2009-04-15 | オリンパス株式会社 | 液浸対物レンズ、蛍光分析装置および倒立型顕微鏡。 |
-
2004
- 2004-01-07 JP JP2004002107A patent/JP2005195878A/ja not_active Withdrawn
-
2005
- 2005-01-05 EP EP05703315A patent/EP1703312A1/en not_active Withdrawn
- 2005-01-05 WO PCT/JP2005/000043 patent/WO2005066683A1/ja not_active Application Discontinuation
-
2006
- 2006-07-07 US US11/482,273 patent/US7274433B2/en not_active Expired - Fee Related
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS59147107U (ja) * | 1982-10-26 | 1984-10-01 | 株式会社富士電機総合研究所 | 先端レンズ損傷防止用保護具 |
JPS59109310U (ja) * | 1983-01-13 | 1984-07-23 | 富士通株式会社 | 顕微鏡 |
JP2003029162A (ja) * | 2001-07-11 | 2003-01-29 | Nikon Corp | 顕微鏡用対物レンズおよび顕微鏡 |
JP2003185926A (ja) * | 2001-12-20 | 2003-07-03 | Nikon Corp | 安全装置付液浸対物レンズ |
Also Published As
Publication number | Publication date |
---|---|
JP2005195878A (ja) | 2005-07-21 |
US20060262419A1 (en) | 2006-11-23 |
US7274433B2 (en) | 2007-09-25 |
EP1703312A1 (en) | 2006-09-20 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US9041940B2 (en) | Three-dimensional shape measuring apparatus | |
JP5779597B2 (ja) | 顕微鏡の構造ユニット、顕微鏡装置及び顕微鏡の構造ユニットの使用 | |
US7274433B2 (en) | Objective, optical analyzer, method of driving optical analyzer, and microscope | |
US20100275334A1 (en) | Modular atomic force microscope | |
KR101488543B1 (ko) | 고침렌즈홀더 | |
JP2003500660A (ja) | レーザスキャナーの走査対象である平面の位置把握方法およびそのためのシステム | |
JP2014507663A (ja) | 統合型顕微鏡ならびに関連する方法および装置 | |
WO2006090593A1 (ja) | 走査型プローブ顕微鏡用変位検出機構およびこれを用いた走査型プローブ顕微鏡 | |
JPH1123219A (ja) | 共焦点光学系による変位計測装置 | |
KR100660112B1 (ko) | 레이저빔의 광폭이 제어되는 레이저와 비전의 동축가공장치 | |
JP4914537B2 (ja) | 倒立型顕微鏡に適用される試料支持装置 | |
JP6985264B2 (ja) | 対物レンズ交換装置を有する顕微鏡 | |
JP5133709B2 (ja) | レーザリペア装置 | |
JP2008134190A (ja) | カンチレバーホルダーおよびこれを備えた走査型プローブ顕微鏡 | |
JPH10267948A (ja) | 走査型プローブ顕微鏡 | |
JP2011112880A (ja) | 顕微鏡 | |
JP2001228071A (ja) | 表面測定器と組み合わされた走査型プローブ顕微鏡 | |
JP2005043892A (ja) | 共焦点ラスタ顕微鏡 | |
CN219574424U (zh) | 透镜耦合设备及半导体激光器生产系统 | |
JP3908178B2 (ja) | 光モジュールの調整装置 | |
CN107247160B (zh) | 一种基于原子力探针的显微镜头与样品台锁定系统 | |
JP2000234994A (ja) | 走査プローブ顕微鏡におけるカンチレバー変位測定方法 | |
KR20160071788A (ko) | 레이저 가공 장치 및 그 구동 방법 | |
CN113588221A (zh) | 一种数值孔径测试系统 | |
CN117092373A (zh) | 紧凑型上扫描式原子力显微镜测头及其使用方法 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AK | Designated states |
Kind code of ref document: A1 Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BW BY BZ CA CH CN CO CR CU CZ DE DK DM DZ EC EE EG ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NA NI NO NZ OM PG PH PL PT RO RU SC SD SE SG SK SL SY TJ TM TN TR TT TZ UA UG US UZ VC VN YU ZA ZM ZW |
|
AL | Designated countries for regional patents |
Kind code of ref document: A1 Designated state(s): BW GH GM KE LS MW MZ NA SD SL SZ TZ UG ZM ZW AM AZ BY KG KZ MD RU TJ TM AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IS IT LT LU MC NL PL PT RO SE SI SK TR BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG |
|
121 | Ep: the epo has been informed by wipo that ep was designated in this application | ||
WWE | Wipo information: entry into national phase |
Ref document number: 2005703315 Country of ref document: EP |
|
WWE | Wipo information: entry into national phase |
Ref document number: 11482273 Country of ref document: US |
|
NENP | Non-entry into the national phase |
Ref country code: DE |
|
WWW | Wipo information: withdrawn in national office |
Country of ref document: DE |
|
WWP | Wipo information: published in national office |
Ref document number: 2005703315 Country of ref document: EP |
|
WWP | Wipo information: published in national office |
Ref document number: 11482273 Country of ref document: US |