WO2010096786A2 - Optical tomography system with high-speed scanner - Google Patents
Optical tomography system with high-speed scanner Download PDFInfo
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- WO2010096786A2 WO2010096786A2 PCT/US2010/024959 US2010024959W WO2010096786A2 WO 2010096786 A2 WO2010096786 A2 WO 2010096786A2 US 2010024959 W US2010024959 W US 2010024959W WO 2010096786 A2 WO2010096786 A2 WO 2010096786A2
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- Prior art keywords
- lens
- scan
- interest
- power element
- variable power
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- 238000003325 tomography Methods 0.000 title claims description 29
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Classifications
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/02—Objectives
- G02B21/025—Objectives with variable magnification
-
- 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
- G01N21/47—Scattering, i.e. diffuse reflection
- G01N21/4795—Scattering, i.e. diffuse reflection spatially resolved investigating of object in scattering medium
-
- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/0004—Microscopes specially adapted for specific applications
- G02B21/002—Scanning microscopes
-
- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B27/00—Optical systems or apparatus not provided for by any of the groups G02B1/00 - G02B26/00, G02B30/00
- G02B27/0075—Optical systems or apparatus not provided for by any of the groups G02B1/00 - G02B26/00, G02B30/00 with means for altering, e.g. increasing, the depth of field or depth of focus
Definitions
- the present invention relates to optical tomographic imaging systems in general, and, more particularly, to a high-speed focal plane scanner for optical projection tomography, in which a small object, such as a biological cell, is imaged by a microscope.
- Background Recently advances in imaging biological cells using optical tomography invented by A. C. Nelson have been disclosed, for example, in U.S. Patent No. 6,522,775, issued 2/18/2003, and entitled "Apparatus And Method For Imaging Small Objects In A Flow Stream Using Optical Tomography," the full disclosure of which is incorporated by reference. Further developments in the field are taught in Fauver et al., US Patent application number 10/716,744, filed 1 1/18/2003 and published as U.S. Publication No.
- Processing in such an optical tomography system begins with specimen preparation.
- specimens taken from a patient are received from a hospital or clinic and processed to remove non-diagnostic elements, fixed and then stained.
- Stained specimens are then mixed with an optical fluid, inserted into a micro- capillary tube and images of objects, such as cells, in the specimen are produced using an optical tomography system.
- the resultant images comprise a set of extended depth of field images from differing perspectives called "pseudo-projection images.”
- the set of pseudo-projection images can be reconstructed using backprojection and filtering techniques to yield a 3D reconstruction of a cell of interest.
- the 3D reconstruction then remains available for analysis in order to enable the quantification and the determination of the location of structures, molecules or molecular probes of interest.
- An object such as a biological cell may be labeled with a stain or tagged molecular probe, and the measured amount and location of this probe may yield important information about the disease state of the cell, including, but not limited to, various cancers such as lung, breast, prostate, cervical and ovarian cancers.
- the depth of field of the imaging optics is extended by scanning an objective lens transverse to a capillary tube containing a specimen.
- a piezoelectric transducer (PZT) actuator moves the objective lens sinusoidally several times per second in order to scan a series of focal planes though a specimen.
- PZT actuator moves the objective lens sinusoidally several times per second in order to scan a series of focal planes though a specimen.
- a focal plane moving through the specimen has its speed limited by inertia inherent in moving the objective lens rapidly along the optical axis through the specimen. Because the objective lens is housed in a relatively massive assembly, the scan rate is typically, about 10 Hz with a theoretical upper limit of roughly 60 cycles per second.
- an image sensor acquires at least one pseudo-projection image.
- an image can be acquired on the down-stroke as well as the up-stroke of the PZT actuator, allowing up to 120 images per second to be acquired. While this is a useful acquisition rate, it can be significantly improved through the apparatus, systems and methods disclosed herein.
- An optical projection tomography system and method for imaging an object of interest An object of interest is illuminated within the field of view of a microscope objective lens located to receive light passing through the object of interest. Light transmitted through the microscope objective lens impinges upon a variable power element. The variable power element is driven to scan through multiple focal planes in the object of interest. Light transmitted from the variable power element is sensed by a sensing element or array.
- FIG. 1 schematically shows an example of a design for an optical tomography image acquisition system with a variable power element for high speed scanning.
- FIG. 2 schematically shows a more detailed example of a design for a variable power element and imaging optics.
- FIG. 3 schematically shows another example of a design for an optical tomography image acquisition system with a variable power element for high speed scanning.
- FIG. 4 schematically shows another example of a design for a variable power element.
- FIG. 5 schematically shows another example of a design for an optical tomography image acquisition system with a variable power element and optical relay.
- FIG. 6 schematically shows an alternate example of a design for an optical tomography image acquisition system with a liquid lens variable power element.
- FIG. 7 schematically shows another example of a design for an optical tomography image acquisition system incorporating a quarter wave plate.
- identical reference numbers identify similar elements or components.
- the sizes and relative positions of elements in the drawings are not necessarily drawn to scale.
- the shapes of various elements and angles are not drawn to scale, and some of these elements are arbitrarily enlarged and positioned to improve drawing legibility.
- the particular shapes of the elements as drawn are not intended to convey any information regarding the actual shape of the particular elements, and have been solely selected for ease of recognition in the drawings. Detailed Description of the Preferred Embodiments
- Capillary tube has its generally accepted meaning and is intended to include transparent microcapillary tubes and equivalent items with an inside diameter generally of 500 microns or less.
- Delivery of field is the length along the optical axis within which the focal plane may be shifted before an unacceptable image blur for a specified feature is produced.
- Object means an individual cell, particle, item, thing or other entity.
- Pulseudoprojection includes a single image representing a sampled volume of extent larger than the native depth of field of the optics.
- One concept of a pseudoprojection is taught in Fauver 744.
- specimen means a complete product obtained from a single test or procedure from an individual patient (e.g., sputum submitted for analysis, a biopsy, or a nasal swab).
- a specimen may be composed of one or more objects. The result of the specimen diagnosis becomes part of the case diagnosis.
- Sample means a finished cellular preparation that is ready for analysis, including all or part of an aliquot or specimen.
- An optical tomography system 100 includes an illuminator 15, a carrier 1 1 , a variable power element 33, and a microscope objective assembly 10.
- a light sensor 30 is located in the focal plane of a tube lens 23 that, in turn, is located to receive light beams 13 transmitted from the variable power element 33.
- image signals acquired by the light sensor 30 are transmitted to a computer or like device for storage and image processing including 3D reconstruction.
- the illuminator 15 is located to illuminate a portion of carrier 1 1.
- the microscope objective assembly 10 is located to receive incident light 5 passing through the carrier 1 1 and objects residing in the carrier 1 1.
- the carrier 1 1 may be coupled to a rotational motor as signified by reference arrow 17. Under control of the rotational motor, the carrier 1 1 may be rotated to present various views of a transported object to the microscope objective assembly 10.
- the carrier 1 1 may advantageously comprise an optically transparent capillary tube or equivalent filled with an optical fluid selected for matching the refractive properties of the optics.
- the variable power element 33 and microscope objective assembly 10 may be aligned along an optical path 12. Light 21 transmitted through the objective lens 10 impinges on the variable power element 33.
- the light sensor 30 may advantageously comprise a photosensor array, a CCD array or equivalents.
- the illuminator 15 may comprise any suitable illumination source or combination of a plurality of illumination sources including sources producing visible light, near infrared, ultraviolet, and other light frequencies useful for optical tomography of fixed or live biological cells and equivalents.
- a variable power element 33 includes a beamsplitter 18, a scan lens 16 and a scan mirror 14 having a reflecting surface 19.
- a light sensor 30 is located in the focal plane of a tube lens 23 that, in turn, is located to receive light beams 13 reflected from one surface of the beam splitter 18.
- the scan mirror 14 is coupled to be moved in a controlled manner by drive 1 14.
- the beam splitter 18, scan lens 16 and scan mirror 14 are aligned along an optical path 12.
- the beam splitter 18 is also tilted at an angle for directing reflected light 13 from the scan mirror 14 through the tube lens 23, ultimately transmitted onto the light sensor 30 for acquiring images of the tube contents.
- Operating the scan mirror 14 comprises oscillating the scan mirror 14 along the optical axis while the reflecting surface 19 of the mirror maintains a substantially perpendicular relationship to the optical axis. As the scan mirror oscillates along the optical axis 12, a plurality of focal planes in the carrier 1 1 (shown in FIG. 1 ) are traversed and the resultant pseudo-projection is imaged on the light sensor 30.
- FIG. 3 another example of a design for an optical tomography image acquisition system with a variable power element for high speed scanning is schematically shown.
- An illuminator 15, a carrier 1 1 , a variable power element 33A, and a microscope objective assembly 10 are aligned along an optical path 12, where the optical path 12 may be the optical axis of the microscope objective assembly 10.
- the microscope objective assembly 10 includes an exit pupil E.
- the variable power element 33A includes a first lens 312 and a second lens 314.
- the second lens 314 may advantageously comprise a scanning lens that is driven along the optical axis 12 by drive 1 14A.
- the exit pupil E, first lens 312 and second lens 314 are each separated by a focal length f. Movement of the second lens 314 operates to translate the second lens by a scanning distance ⁇ to scan focal plans in the carrier 1 1.
- the scanning distance ⁇ will oscillate between a positive and negative value represented here as f + ⁇ .
- variable power element 33B includes a variable focal lens 412.
- the variable focal lens 412 may be comprised of a transparent cell filled with two non-miscible fluids made according to "Varioptic" technology.
- Varioptic lenses are based on an electrowetting principle and are controllable using a variable voltage source 1 14V.
- Varioptic lenses are commercially available from Varioptic SA, Lyon, France. Such lenses can present large optical power variations.
- a 5mm diameter Varioptic lens can have an inverse focal length that varies up to 50 dioptries. (See, e.g., Gabay, et al. "Dynamic Study of a Varioptic variable focal lens," Current Developments in Lens Design and Optical Engineering III, Proceeding of SPIE Vol. 4767 (2002)).
- FIG. 5 another example of a design for an optical tomography image acquisition system with a variable power element and optical relay is schematically shown.
- a microscope objective assembly 10, an optical relay 510 and a variable power element 33C are aligned along an optical path.
- the microscope objective assembly 10 includes an exit pupil E.
- the variable power element 33B may include a beamsplitter 18 and a scan lens 16.
- the optical elements are each separated by a focal length f.
- E represents the entrance pupil of the scan lens 16 which is coincident with the entrance pupil of the variable power element 33C.
- Broken line 516 indicates the light path for focusing the exit pupil E to be coincident with the entrance pupil E'.
- Light rays 521 represent light transmitted through the objective lens 10.
- Light rays 523 represent light relayed by optical relay 510.
- the beam splitter 18 is configured for directing reflected light 13 from the scan lens 16 and scan mirror 14 through the tube lens 23, ultimately transmitted onto the light sensor 30 for acquiring images of the tube contents.
- the optical relay 510 includes a first focal lens 512 aligned with a second lens 514, where the first and second lenses are spaced by a distance of twice the focal length, 2f. Having described the structure of example embodiments, a description of the operation of example embodiments should prove helpful to further understanding of the principles of the disclosure. Translating the scan mirror 14 a distance Az' produces a focus shift ⁇ z at the object equal to 2 Az' n I (n !
- n is the object space index of refraction
- n' is the scanner space index of refraction
- m r is the transverse magnification between the object and scanner.
- the factor of 2 is due to the fact that the optical path length changes by twice the mirror translation.
- the mirror need be scanned by only 1.5 microns.
- the scan mirror need only be as large as the scanner space field of view, which is typically less than 1 mm.
- a scan mirror 14 having a 1 mm diameter would be sufficient to reflect the entire image.
- Such a mirror weighs much less than a typical objective lens, and scanning such a mirror takes much less force and energy than does scanning the objective lens.
- the lower required force and shorter scan distance allow scan frequencies that are thousands of times greater than are feasible with a scanning objective.
- the scan mirror 14 and drive 1 14 may be made from a wide variety of available fabrication technologies.
- One example is a conventional first surface mirror attached to a piezoelectric actuator.
- Another example is a micro-electro-mechanical- systems (MEMS) flexure with a reflective coating.
- MEMS micro-electro-mechanical- systems
- Yet another example is a metal mirror with a solenoid actuator.
- An acoustic wave generator driving a reflective elastic diaphragm may also be used.
- One skilled in the art will recognize that many other combinations of conventional actuators and reflectors may also be used.
- the scan magnification AzI ⁇ z' is independent of the numerical aperture (NA) of either the objective lens 10 or scan lens 16, but the NAs of the two lenses are indirectly tied to the scan magnification.
- NA numerical aperture
- the entrance pupil E', of the scan lens 16 must be large enough to avoid vignetting in order to realize the resolving power of the objective lens 10 over the entire field of view.
- the scan lens 16 NA be equal to or larger than the objective lens 10 NA divided by the transverse magnification m ⁇ , as follows from the optical invariant.
- the factor m r ties the NAs to the scan magnification Az I Az'. Equality is sufficient when the exit pupil E of the objective lens 10, or its image, is coincident with the entrance pupil E' of the scan lens 16, otherwise the scan lens 16 NA must be larger.
- An optical relay may be used to image the exit pupil E of objective 10 onto the entrance pupil E' of scan lens 16. This is necessary if the lens pupils are virtual pupils, as is the case with most high performance high magnification microscope objectives. Moreover, a relay magnification m E of less than one, may be used to reduce the size of the image of the exit pupil of objective lens 10 to the point where it is the same size as the entrance pupil of scan lens 16. Using this technique has the side effect of also reducing the scan magnification by m E .
- the image magnification must remain constant while scanning to obtain a good pseudoprojection image. This is accomplished by making the system telecentric in scanner space. If the scan lens 16 is a telecentric lens, wherein its entrance pupil lies in its front focal plane, then the system will be telecentric in scanner space if and only if the exit pupil of objective lens 10, or its image, is coincident with its entrance pupil. If the scan lens 16 is not telecentric, then placing the exit pupil of objective lens 10, or its image, at the front focal plane of the scan lens 16 will make the system telecentric in scanner space, so long as the objective lens 10 exit pupil is not vignetted by scan lens 16.
- a relay system as described in the previous paragraph, may be used to keep the magnification constant while scanning if either of the pupils is a virtual pupil.
- one advantageous embodiment includes (1 ) A scan lens having as high an NA as possible,
- FIG. 6 an alternate example of a design for an optical tomography image acquisition system with a liquid lens variable power element is schematically shown.
- a microscope objective assembly 10, a variable power element 33B are aligned along an optical path.
- the variable power element 33B includes a variable focal lens 412, for example a Varioptic lens or equivalent, as discussed above with reference to FIG. 4.
- An intermediate lens 611 is located in the optical path between the microscope objective assembly 10 and the variable power element 33B.
- the tube lens 23 is located to receive light beams 13 transmitted from the variable power element 33B and transmit the light beams to the light sensor 30.
- the variable focal lens 412 may be located a distance 2f from the intermediate lens 61 1.
- the intermediate lens 61 1 may advantageously be located at a distance 2ffrom the microscope objective assembly 10.
- An alternate fast scan mirror design 200 employs a polarizing beam splitter 218 optical aligned with a quarter wave plate 220 so as to transmit substantially circularly polarized light.
- the reflected light 13P from the scan mirror is then substantially circularly polarized with opposite handedness.
- the quarter wave plate 220 converts this to linear polarization light 13P' substantially orthogonal to the incident light 5.
- the beam splitter 218 is configured for directing the linear polarization light 13P' from the scan mirror 14 through the tube lens 23, ultimately transmitted onto the light sensor 30 for acquiring images of the tube contents.
- the other elements are similar in type, configuration and construction to those as described above with reference to FIG.1.
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Abstract
Description
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Priority Applications (5)
Application Number | Priority Date | Filing Date | Title |
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CN201080017727.0A CN102428343B (en) | 2009-02-23 | 2010-02-22 | Optical tomography system with high-speed scanner |
EP10744448.1A EP2406580A4 (en) | 2009-02-23 | 2010-02-22 | Optical tomography system with high-speed scanner |
AU2010215794A AU2010215794B2 (en) | 2009-02-23 | 2010-02-22 | Optical tomography system with high-speed scanner |
JP2011551280A JP2012518794A (en) | 2009-02-23 | 2010-02-22 | Optical tomography system with high-speed scanner |
CA2752878A CA2752878C (en) | 2009-02-23 | 2010-02-22 | Optical tomography system with high-speed scanner |
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US12/391,096 | 2009-02-23 | ||
US12/391,096 US8254023B2 (en) | 2009-02-23 | 2009-02-23 | Optical tomography system with high-speed scanner |
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WO2010096786A2 true WO2010096786A2 (en) | 2010-08-26 |
WO2010096786A3 WO2010096786A3 (en) | 2011-02-17 |
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PCT/US2010/024959 WO2010096786A2 (en) | 2009-02-23 | 2010-02-22 | Optical tomography system with high-speed scanner |
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EP (1) | EP2406580A4 (en) |
JP (1) | JP2012518794A (en) |
CN (1) | CN102428343B (en) |
AU (1) | AU2010215794B2 (en) |
CA (1) | CA2752878C (en) |
WO (1) | WO2010096786A2 (en) |
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Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
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US10684456B2 (en) | 2015-10-16 | 2020-06-16 | Olympus Corporation | Microscope |
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CN102428343B (en) | 2014-08-20 |
JP2012518794A (en) | 2012-08-16 |
US8254023B2 (en) | 2012-08-28 |
WO2010096786A3 (en) | 2011-02-17 |
US20100214639A1 (en) | 2010-08-26 |
AU2010215794A1 (en) | 2011-09-22 |
CA2752878C (en) | 2015-12-22 |
EP2406580A4 (en) | 2015-05-13 |
CA2752878A1 (en) | 2010-08-26 |
EP2406580A2 (en) | 2012-01-18 |
AU2010215794B2 (en) | 2014-05-01 |
CN102428343A (en) | 2012-04-25 |
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