EP1842095A2 - Microscope optique de scannage adaptatif - Google Patents

Microscope optique de scannage adaptatif

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Publication number
EP1842095A2
EP1842095A2 EP05855789A EP05855789A EP1842095A2 EP 1842095 A2 EP1842095 A2 EP 1842095A2 EP 05855789 A EP05855789 A EP 05855789A EP 05855789 A EP05855789 A EP 05855789A EP 1842095 A2 EP1842095 A2 EP 1842095A2
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EP
European Patent Office
Prior art keywords
light
image
lens
optics
field
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP05855789A
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German (de)
English (en)
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EP1842095A4 (fr
Inventor
Benjamin Michael Potsaid
Yves Bellouard
John T. Wen
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Rensselaer Polytechnic Institute
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Rensselaer Polytechnic Institute
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Application filed by Rensselaer Polytechnic Institute filed Critical Rensselaer Polytechnic Institute
Publication of EP1842095A2 publication Critical patent/EP1842095A2/fr
Publication of EP1842095A4 publication Critical patent/EP1842095A4/fr
Withdrawn legal-status Critical Current

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Classifications

    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/36—Microscopes arranged for photographic purposes or projection purposes or digital imaging or video purposes including associated control and data processing arrangements
    • G02B21/365—Control or image processing arrangements for digital or video microscopes
    • G02B21/367—Control or image processing arrangements for digital or video microscopes providing an output produced by processing a plurality of individual source images, e.g. image tiling, montage, composite images, depth sectioning, image comparison
    • 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
    • G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B26/00—Optical devices or arrangements for the control of light using movable or deformable optical elements
    • G02B26/06—Optical devices or arrangements for the control of light using movable or deformable optical elements for controlling the phase of light

Definitions

  • the present invention relates generally to the field of optical microscopy and, in particular, to a new and useful adaptive scanning optical microscope which addresses and improves upon, the usual trade-off between resolution and field of view that is common for known optical microscopes.
  • the inventors have disclosed an earlier approach to solving this trade-off problem in U.S. Patent Application No. 10/525,422 filed February 25, 2005.
  • This application claims priority on U.S. Provisional Patent Application No. 60/411 ,038 and International Application No. PCT/US2003/029332, published as WO 2004/025331 , all of which are also incorporated here by reference.
  • For a wide range of applications e.g.
  • the optical microscope remains one of the most important tools for observing below the threshold of the naked human eye.
  • the present invention is a new optical microscope design that combines a scanner lens, a steering mirror, an adaptive optics element, adaptive optics (AO) conditioning optics, and imaging optics to enlarge the field of view while preserving resolving power in the acquired images.
  • This instrument has the ability to operate at high image acquisition rates for increased throughput orto facilitate certain spatial-temporal observations.
  • the present invention achieves an expanded field of view at high resolution by integrating active optical elements, motion control, and image processing techniques with traditional static optical elements in a tightly integrated fashion.
  • SOMS Scanning Optical Mosaic Scope
  • this critical distance occurs when the center of one Airy disk falls on the first minimum of the other and is related to the numerical aperture, NA, of the system and the wavelength of light, ⁇ .
  • NA numerical aperture
  • the NA of the system is a function of the index of refraction of the transmitting medium, n, and the half angle of the cone of light collected from the object.
  • Lithography lenses require near perfect manufacturing and extremely tight assembly tolerances (often requiring an interferometric assembly process), and can. cost in the millions of dollars. Also, negatively powered elements are required and are located at narrow beam regions in both the microscope and lithography lenses and positively powered elements where the beam is wide. This design technique is used to achieve a flat imaging field (small Petzval sum) and results in an increase in the lens count and optical complexity. An additional consideration is the size of the image sensor, given that large commercially available CCD cameras only have approximately 9216 x 9216 pixels (e.g. Fairchild Imaging CCD595).
  • the "basic post-objective scanning” method refers to the commercially available units, which are limited to very low numerical aperture and suffer from considerable off-axis aberration because of the system layout.
  • Dmetrix is covered by several patents, for example: US Patent 6,958,464, for an Equalization for a multi-axis imaging system; US Patent 6,950,241 for a Miniature microscope objective for an array microscope; US Patent 6,905,300 for a Slide feeder with air bearing conveyor; and US Patent 6,842,290 for a Multi-axis imaging system having individually-adjustable elements.
  • This system uses an array of 80 miniature microscopes (each of 3 element aspheric design) working in parallel to rapidly acquire the image.
  • a related technology is the line scanning system, which sweeps a specimen (often projected through a microscope objective) past a linear array of sensor pixels.
  • a major disadvantage of line scanning technology is that images are obtained line by line (n x 1 pixels) as opposed to area by area (n x n pixels), as is the case with a more typical area based image sensor.
  • line scan systems generally require extremely short exposure times and/or bright illumination to obtain high throughput, which is often not possible in biological applications where photo-damage, bleaching, and fluorescence must be considered.
  • the DMetrix excels at static and high fill factor applications.
  • Fill factor is the percentage of the total observable area that is of interest and absolutely must be imaged or sensed for the application at hand.
  • the ASOM of the present invention acquires images serially in time with extremely fast repositioning speeds, the ASOM will excel in dynamic and/or low fill factor applications.
  • Low fill factor applications include biological imaging of rare events over a large cell population, tracking multiple moving organisms, medical diagnostics of tissue sampled by needle extraction which is haphazardly placed on a microscope slide, etc. Most manufacturing applications require a low fill factor as only certain critical regions need to be observed or inspected with dynamic tracking of objects or features often required during assembly.
  • the ASOM of the present invention is particularly suitable for challenging spatial-temporal observation tasks requiring both a wide field of view and high resolution. Consideration of these issues motivated and contributed to the design of the ASOM.
  • deformable mirrors have also been used to correct for off-axis aberrations and sample induced wavefront disturbances in confocal microscopy.
  • a confocal microscope of know design uses a pinhole screen located in a plane that is conjugate to the object plane. This pinhole rejects light that is not at the same depth as the focal plane. The pinhole also rejects light that is not at the center of the field. Thus, the confocal microscope samples the object point by point. The image is built up point by point and there is a means for scanning the location of the imaging point on the specimen.
  • a basic introduction to the confocal microscope can be found at: http://www.physics.emory.edu/ ⁇ weeks/confocal.
  • the ASOM of the present invention acquires images using finite imagery
  • US Patent 6,771 ,417 discloses a non-confocal arrangement that includes adaptive optics. See US Patent 6,555,826 for a confocal arrangement including adaptive optics and US Patent 6,381 ,074 for an adaptive optics element in a scanning confocal microscope to assist in aberration control and precise focusing. US Patent 6,483,641 discloses a spatial light modulator used in a microscope. SUMMARY OF THE INVENTION
  • the ASOM design of the present invention shares the scanning and mosaic construction principle of the SOMS (again see U.S. Patent Application No. 10/525,422), however, the ASOM of the present invention differs from the SOMS and related post-objective systems performing finite imagery in that the ASOM incorporates an adaptive optics element to address off-axis aberrations introduced by a custom designed scanner lens that allows for aberrations not achieving the diffraction limit. Additionally, the ASOM differs from existing technologies in that the scanner lens is simplified by relaxing the flat field requirement and works with the steering mirror to project a significantly curved intermediate image field that rotates about its own center. Using these ideas to simplify the optical complexity and reduce the manufacturing and assembly requirements, the underlying concept of the novel ASOM is to use a low mass and very fast steering mirror located between the scanner lens and the imaging optics to form a post-objective scanning configuration.
  • an image is acquired and the associated light advanced along an optical path that includes an adaptive optics element (e.g. a deformable mirror, a spatial light modulator, an optical phased array, a deformable lens, or similar optical element) to correct for the residual aberrations that are scan position dependant.
  • an adaptive optics element e.g. a deformable mirror, a spatial light modulator, an optical phased array, a deformable lens, or similar optical element
  • the ASOM design of the present invention excels in applications requiring high throughput, relatively low lighting conditions, and/or critical spatial-temporal observations, but will not offer the virtually unlimited field of view associated with a moving stage.
  • Biological applications where the ASOM would be attractive include observing dynamic cellular events (mitosis, viral attachment, motility, cellular response to chemical application) over a large population of living cells or observation of select regions of interest on samples.
  • the ASOM would also be useful for rapidly acquiring images from well plates or for providing vision feedback in micro-injection or manipulation activities. By injecting light into the optical path and installing appropriate filters, epi-illumination modes would allow fluorescent imaging. By installing a phase plate near the aperture, phase contrast imaging could also be achieved for observation of predominately phase objects as is common in biology.
  • the ASOM allows for vision guided micro-assembly, processing, and rapid inspection of parts, with the potential for higher product throughput.
  • the ASOM allows for rapid imaging of biological samples. For example, in the case of biopsies obtained with a needle extraction, the sample is placed haphazardly on the slide and occupies only a small portion of the slide area.
  • the ASOM will be able to perform a very rapid background scan and then a high quality scan of only the biopsy region of interest.
  • the high speed of the rapid background scan will be obtained by imaging without stopping the steering mirror motion. These images will be slightly blurred, but will allow the tissue sample location to be identified.
  • the ASOM will then plan a scanning trajectory to capture the region of interest.
  • a further objective of the present invention is to provide an adaptive scanning optical microscope which comprises a scanner lens assembly for acquiring images from different parts of an object plane and for forming a preferably curved image field having at least some aberration which varies as a function of the part of the object plane from which the image is acquired, a steering mirror for steering light from the image field and along a light path from the object plane to an final image plane, an adaptive optics component for receiving the steered light from the object plane to the image field and for dynamically compensating for the aberration, and additional optics along at least part of the light path for guiding, conditioning, and focusing the light as it moves from the steering mirror, past the adaptive optics element and to the final image plane.
  • Fig. 1 is a schematic conceptual illustration of the adaptive scanning optical microscope or ASOM of the present invention
  • Fig.2 is a composite diagram showing the various conjugate image and aperture planes of the invention.
  • Fig. 3 is a multi-part illustration showing, at (a) the shape of the image field for a thin lens, at (b) the curved surface of the retina (image sensor) that allows for a very simple lens in the human eye, and at (c) the ASOM scanner lens of the present invention which is simplified by allowing a curved image field;
  • Fig.4 is a composite illustration of the curved image field of the scanner lens assembly of the present invention;
  • Fig.5 is a composite illustration of the field curvature of the scanner lens assembly and imaging optics of the invention.
  • Fig.6 is an illustration showing a preliminary design of the ASOM of the present invention.
  • Fig. 7 shows a 40mm virtual field of view of the ASOM of the present invention, as compared to that offered by a traditional microscope using 1024x1024 and 4096x4096 cameras (all systems operating at 0.21 NA), the 0.38 mm size of the ASOM sub-field of view being also shown with a 512x512 camera, requiring many scan movements to cover the entire 40mm field;
  • Fig. 8 is a diagram that shows some of the different operating modes of the ASOM of the present invention.
  • Fig. 9 is a composite view illustrating, at (a), different field positions of the invention, at (b), optimal deformable mirror shape for each specific field position and at (c), Strehl ratios sampled over the selected field of view; and
  • Fig. 10 is an optical layout of the inventors' latest experimental setup for demonstrating the principles of the present invention.
  • Fig. 1 shows the adaptive scanning optical microscope or ASOM 10 which operates by taking a sequence of small spatially displaced images in succession from an object 12, and then assembling a large composite image (mosaic) or several disjoint or possibly overlapping images of the scene.
  • ASOM 10 the adaptive scanning optical microscope or ASOM 10 which operates by taking a sequence of small spatially displaced images in succession from an object 12, and then assembling a large composite image (mosaic) or several disjoint or possibly overlapping images of the scene.
  • the mechanism and scanning principle in the invention include a high speed 2-D steering mirror 14 working in coordination with a specially designed scanner lens assembly 16, an adaptive optics (AO) element 18 (for example, a deformable mirror, a spatial light modulator, an optical phased array, a deformable lens, or similar optical element), and additional imaging optics 20.
  • AO adaptive optics
  • the image is eventually sampled by a sensor, 22 e.g. a digital camera of suitable quality and speed, a spectrometer, or other light sensitive device.
  • the imaging optics 20 include forward AO (adaptive optics) conditioning optics or eye-piece 32, inverted AO conditioning optics or eye-piece 34 and final imaging optics 36, which may each be made up of one or more elements.
  • forward AO adaptive optics
  • inverted AO conditioning optics or eye-piece 34 and final imaging optics 36, which may each be made up of one or more elements.
  • Fig. 1 also illustrates the at least one electronic system 15 for controlling the position of the steering mirror, and 17 for controlling the actuator signals to the adaptive optics element.
  • At least one electronic system 19 is also provided for reading data from the sensor (e.g. camera 22) and at least one of displaying, processing and/or storing the acquired data of the light at the final image plane.
  • Fig. 2 shows the conjugate image and aperture planes of the ASOM of the present invention, and partitions the optical elements into a scanner lens 16, forward eye-piece 32, inverted eye-piece 34, and final imaging optics 36, these last three optical assemblies or elements forming the additional image optics 20.
  • the scanner lens 16 collects light from the object 12 or object plane 1 while the steering mirror 14, located at an image of the pupil, aims a projected real intermediate image.
  • a first image of aperture A1 follows the SLA 16 which is followed in the light path, by a first intermediate image plane 2.
  • the forward eye-piece 32 in the ASOM 10 samples the first intermediate image 2 and projects an external pupil to where the deformable mirror 18 is located.
  • the forward AO conditioning optics 32 in the preliminary design of the invention resembles a Huygens' eyepiece in that the intermediate image 2 is located between a negative field-lens 33 and a positive eye-lens 35.
  • a notable difference is the use of a negative field lens 33. This has the effect of lengthening the adaptive optics element relief (distance between eye-lens 35 and adaptive optics element e.g., the deformable mirror 18), but at a cost of a larger eye-lens 35.
  • a second image of the aperture A2 follows the eye-lens 35.
  • the inverted AO conditioning optics 34 resembles a Kellner eyepiece, but has a negative field lens 37 that follows the positive lens and the second image of the aperture A2.
  • the negative field lens 37 also helps contribute to a negative Petzval sum in the imaging optics and establishes a second intermediate image plane 3.
  • the inventors did use the Huygens' and Kellner eyepiece like configurations. However, there are many configurations of eye-pieces that would work well here. In the experimental system set up in the lab the invention use eye-piece configurations that contain up to seven lens elements. It is not the type of eye-piece that is important, but the function of the eye-piece that is important to define the ASOM. For this reason, the forward and inverted eye-pieces of the present invention are better described as the forward and inverted AO conditioning optics.
  • the forward eye-piece pupil imaging optics there are many different ways to construct the forward eye-piece pupil imaging optics, the inverted eye-piece pupil imaging optics and final imaging optics.
  • the simulated design discussed here uses two lens elements for the eye-pieces and one lens element for the final imaging optics.
  • the experimental setup in the lab uses seven elements in the forward eyepiece, three in the inverted eyepiece, and seven in the final imaging optics.
  • the person of ordinary skill in the art of optics once understanding the principles of the present invention, will be able to assemble other embodiments of the invention.
  • the final imaging optics 36 relay the second intermediate image 3 to the sensor (e.g. the science camera 22 - see Fig. 1) at the final image plane 4 with the proper magnification to prevent aliasing.
  • the system aperture stop defines the boundary of the ray bundles accepted by the imaging system.
  • the active area of the sensor will provide for a field stop, but an additional field stop can be added at the first and second intermediate image fields to reduce stray and unwanted light in the system.
  • Other baffling and stray light reducing mechanisms such as machined grooves in the mechanical housings and a black coating applied to the surfaces would likely be used throughout the ASOM optical path.
  • the present invention also advantageously uses a curved field scanning layout that is different from a microscope objective or lithography lens.
  • the scanner lens 16 of the ASOM 10 of the present invention is designed to exhibit significant field curvature C with a relatively large Petzval sum.
  • This relaxation of the flat field requirement offers the advantage of a greatly simplified optical design with far fewer lens elements, as the "natural" behavior of a lens is to image with a curved image field as shown in Fig. 3, area (a) for thin lenses and in area (b) for the human eye.
  • the shape of the projected image surface is nearly spherical instead of the more typical parabolic surface associated with field curvature. This is achieved through higher order aberration control.
  • This layout is advantageous because it eliminates the need for a large and flat field imaging system. Instead, as shown in Fig. 5, the system exhibits (1) a large positively curved field associated with the scanner lens, and (2) a small negatively curved field associated with the imaging optics, thus avoiding the significant difficulty of designing and manufacturing a large continuous flat field imaging system as discussed above.
  • the imaging optics are low numerical aperture, small field size, and used predominantly on-axis, the inventors have found that off-the-shelf optics can provide sufficient aberration correction for diffraction limited performance when used with medium size sensor arrays (512 x 512 pixels). Larger sensor arrays may require custom imaging optics.
  • Fig.6 the adaptive optics element wavefront correction of the present invention will be described.
  • the present invention circumvents this problem by designing a "good" scanner lens with significant wavefront aberration (up to several waves of optical path difference), and then uses a deformable mirror as the adaptive optics element 18 to compensate for the aberrations over the specific viewing field that is selected. Variation in the aberration is allowed between individual field positions throughout the scanner's range. However, given that the deformable mirror can only achieve one specific shape at a time, the rate of change in the aberration between field positions must be small enough to allow diffraction limited imaging performance over the entire sub-field of view that is selected.
  • Fig. 7 compares the observable field of view of the ASOM to a fixed microscope with a 4096 x 4096 camera (considered a full field camera with standard microscope objectives) and with a 1024 x 1024 camera, which is more common.
  • the ASOM offers diffraction limited (Strehl ratio > 0.8) for all field positions based on high fidelity simulation.
  • Fig. 7 Also shown in Fig. 7 is the sub-field of view offered by the 512 x 512 camera used in this ASOM implementation.
  • the relatively simple imaging optics limit the camera sensor size to be about 6.0 mm in diameter for diffraction limited performance.
  • This also shows the performance of only one specific implementation of the ASOM.
  • the field size and numerical aperture can be tailored to the observation task at hand. However, in general, there will ultimately be a tradeoff between the maximum observable field size and the numerical aperture of the system.
  • Fig. 8 illustrates different operating modes of the ASOM of the present invention, such as rare event detection, tracking moving objects in time, imaging only regions of interest and full area coverage.
  • Fig. 9 shows how the DM corrects for the specific wavefront aberration associated with each field position. Over the entire field and for all field positions, the Strehl ratio is much greater than the diffraction limit of 0.8, resulting in near perfect imaging. Area (a) illustrates five different field positions. Area (b) illustrates the five corresponding optimal deformable mirror shapes for each respective field position and area (c) gives the Strehl ratio sampled over the selected field of view. [0073] All results presented here are based on idealized simulations ignoring the reality that lenses and optical housings are always subject to manufacturing and assembly tolerances.
  • a Micro-assembly demonstration of the invention is based on a shape memory alloy micro-gripper moving between two fixed objects in a workspace.
  • a rudimentary correlation based image matching algorithm and Kalman filter are used to track the motion of the gripper tip.
  • a 3 x 3 tile mosaic images the gripper and the scanning pattern is automatically adjusted to maintain the gripper tip in the center tile.
  • the scan pattern also includes the two stationary objects in the workspace, demonstrating the capability of the SOMS to observe multiple stationary and moving objects in the workspace nearly simultaneously.
  • a sequence of video footage was taken, specifically of living biological cells (Telomerase-lmmortalized hTERT-RPE1 ).
  • a 3 x 3 tile image mosaic monitors a large cell population without disturbing the cells, which are kept alive in a temperature regulated nutrient solution.
  • Several events of mitosis can be seen occurring throughout the viewing field.
  • the ASOM not only offers the possibility of automatically detecting the onset of mitosis and other events, but can be easily programmed to track and record multiple events at the same time. While automated quantitative cell analysis using a moving stage has recently been proposed, the bandwidth of the overall system is still constrained by the response of the stage and the sensitivity of the cell specimen to motion.
  • the inventors have also build a second generation experimental prototype.
  • this experimental apparatus has been carefully designed to demonstrate the critical optical characteristics that define the ASOM, including the curved field optical scanning approach and wavefront correcting optics using a deformable mirror as the adaptive optics element.
  • the steering mirror is manually actuated, limiting the microscope to observing static or slow moving objects.
  • a commercial version of the adaptive scanning optical microscope would likely utilize custom manufactured optics to fully realize the potential of the ASOM concept to achieve higher numerical aperture and a larger workspace as well as incorporate an actuated high speed steering mirror.
  • Fig. 10 shows the optical layout for this experimental system.
  • This initial prototype utilizes a transmitted lighting scheme and because the current design is very sensitive to chromatic aberration, a 510nm wavelength notch filter is used to eliminate much of the light spectra below 500nm and above 520nm.
  • Light transmits through the object contrast pattern and is then collected by the telecentric twelve element scanner lens assembly, which projects an image of the object onto a spherically curved image field.
  • a manually actuated steering mirror with kinematics that pivot the mirror about its silvered front surface is located after the scanner lens assembly, and working in coordination with a field stop in the wavefront correcting optics, selects which portion of the spherically curved image field passes through the system to form an image at the camera.
  • This scanning mechanism effectively allows for the steering of the sub-field of view within the workspace.
  • the light at this point exhibits significant wavefront aberration as a result of the poor optical correction of the scanner lens (note that allowing for poor correction is a characteristic of the ASOM design as it significantly reduces the complexity and lens count of the scanner lens assembly).
  • a MEMS deformable mirror is used in this embodiment of the adaptive scanning optical microscope. By precisely controlling the shape of the reflective surface of the mirror to be opposite the shape of the wavefront error (but at half the amplitude), the deformable mirror can correct for the wavefront aberrations to within the diffraction limit. Thus light leaving the deformable mirror is well corrected and will form an image on the camera that is nearly indistinguishable from a perfect diffraction limited image.
  • a three layer MEMS deformable mirror available from Boston Micromachines Corp. was used in this prototype.
  • This mirror has 32 electrostatic actuators with 400 ⁇ m actuator spacing, a 2.5 ⁇ m actuator stroke, and a 2.0 mm diameter actively controlled area.
  • the 2.5 ⁇ m stroke is capable of correcting for several waves of aberration, which allows for high image quality even for the off-axis field positions, enabling the greatly expanded field of view in the ASOM.
  • the preconditioning stage for the adaptive optics element and the post-conditioning stage for the adaptive optics element are comprised of a suitable combination of positive and negative lens elements to condition the light to match the 2.0 mm active diameter of the adaptive optics element.
  • the pre-conditioning stage for the adaptive optics element forms an image of the aperture such that the steering mirror is located in the vicinity of the aperture image.
  • Placing the steering mirror at or near the aperture image allows for the diameter of the steering mirror to be made small to reduce the inertia of the steering mirror for faster dynamic performance. Additionally, placing the steering mirror at or near the aperture image is optically desirable because doing so facilitates symmetric use of the scanner lens during scanning (i.e. the chief rays for all field positions originate from the same location in the aperture plane).
  • the optical benefits of using a single steering mirror are well know, but for a variety of reasons, including cost and dynamic performance, it is conceivable that the single steering mirror described here would be replaced by two single axis steering mirrors. Doing so is less desirable from an optical standpoint, but is often adequate and is quite often done in practice.
  • An ASOM could be constructed using two or more rotating mirrors.
  • the adaptive optics element is located in the vicinity of an image of the aperture. Locating the adaptive optics element at or in the vicinity of an image of the aperture makes effective use of the active region of the adaptive optics element because all of the ray bundles overlap at this location (i.e. the chief rays cross the optical axis). Locating the adaptive optics element at or near where the chief rays intersect also allows one adaptive optics wavefront correction (e.g. deformable mirror shape) to effectively correct for the aberrations in each ray bundle, as they are similar across each instantaneous field of view.
  • one adaptive optics wavefront correction e.g. deformable mirror shape
  • the ultimate image quality of the adaptive scanning optical microscope depends on the magnitude and shape of the residual wavefront aberrations in the system. Additionally, the scanner lens introduces aberrations specific to each field position (steering mirror angle). Thus, given that the adaptive optics element influence on the wavefront shape is controlled by the control signals and has a direct effect on the wavefront aberrations, an initial calibration of the adaptive scanning optical microscope should be performed. The goal of this calibration is to find a set of control signals that minimize the wavefront aberrations for different field positions. Once calibrated, the optima! control signals can be recalled during normal operation, possibly from a lookup table and using interpolation.
  • This calibration can compensate for manufacturing and assembly errors, tolerances, or other variations in manufacturing, and may be periodically repeated to compensate for changes in environmental temperature, shifting or changing of optical components, or other sources of aberration in the system.
  • the steering mirror and adaptive optics element would likely be coordinated by an electronic system for effective compensation of optical aberrations during imaging.
  • the optimal adaptive optics control signals there are many potential methods to obtain the optimal adaptive optics control signals, including using a wavefront sensor, wavefront estimating experimental methods and algorithms, interferometer based methods, or other image based technique. It is also conceivable that the adaptive scanning optical microscope system would update the adaptive optics element's control signals during runtime using a real-time measurement of the wavefront aberrations and feedback control similar to as is done with adaptive optics telescopes using a guide star as a reference wavefront. [0088] For this experimental prototype, a performance metric and a numerical optimization algorithm were used; In general, the performance metric, Q(u), is a nonlinear function of adaptive optics control signals, u, and Q(u) is defined to decrease with improving image quality.
  • the resulting optimization problem is also subject to upper and lower bounds on the adaptive optics control signals.
  • Combining a metric based on the high frequency image content and the parallel stochastic-gradient-descent (PSGD) optimization algorithm has been demonstrated to effectively calibrate the system.
  • the adaptive optics control signal optimization requires two parts for which there are a variety of possible options and specific combinations:
  • Additional enhancements to the ASOM described above include using a spectrometer in place of the camera. Introducing a phase plate near the aperture and/or using the adaptive optics element to introduce a phase perturbation to the wavefront would allow the ASOM to perform phase contrast imaging. Light could also be injected into the ASOM as a means to illuminate the object, possibly by introducing a beam splitter into the optical path.
  • the present invention is a new microscope concept that can simultaneously achieve high resolution and a large effective field of view that offers several advantages over the current state of the art for observing certain spatial-temporal events.
  • the design draws heavily on the synergy of an optical, mechanical, motion control, and image processing design.
  • ZEMAX optical simulations show diffraction limited imaging performance over a greatly enlarged field of view, while calculations show the. possibility for high speed movement and image acquisition operation.
  • a reduced functionality proof-of- concept prototype has been constructed to demonstrate the basic efficacy of the mirror based scanning approach and we demonstrate with both micro-assembly and biological observation tasks.
  • the scanner lens assembly, the forward and inverted conditioning optics, and the final imaging optics of the invention can each be constructed of one or more glass lens elements; plastic lens elements; GRIN (graduated index of refraction) elements; diffractive lens elements; spherical optical elements; aspherical optical elements; elements or collection of elements exhibiting an external pupil, a telecentric behavior, a non-telecentric behavior; a uniform numerical aperture for all field positions, a non-uniform numerical aperture for different field positions; an element or collection of elements that substantially obeys an f-theta distortion mapping, an f-cosine-theta distortion mapping, an f- sine-theta distortion mapping; an element or collection of elements that projects a curved image field at a first intermediate image plane, a curved image field at the first intermediate image plane that is substantially spherical, and a curved image field at the first intermediate image plane that is substantially parabolic.
  • GRIN graduated index of refraction
  • the image steering mirror can be generalized as any image steering means or equivalent for performing the steering function, and these means may include means for steering including at least one galvanometer, voice coil actuator, piezo electric actuator, electrostatic actuator, gimbal mechanism, a parallel mechanism, a flexure mechanism, or a electro-magnetic levitation.
  • the steering mirror may be at least one of a flat reflecting surface, a curved reflecting surface, a curved reflecting surface that is substantially spherical, a curved reflecting surface that is substantially aspherical, or a rotating prism.
  • the sensor for receiving light at the final image plane can be at least one of: a digital camera, a charged coupled device, a CMOS sensor, a spectrometer, or an eyepiece for view with the human eye.

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  • Lenses (AREA)

Abstract

L'invention concerne un microscope optique de scannage adaptatif qui comprend un ensemble de lentilles de scanneur permettant d'acquérir des images de différentes parties d'un plan d'objet et de former un champ d'image courbé possédant au moins certaine aberration qui varie comme une fonction de la partie du plan d'objet, à partir duquel l'image est acquise. Un miroir de guidage permet de sélectionner le champ de vue et de guider la lumière à partir de l'objet et le long d'une voie de lumière du plan de l'objet à un plan d'image final. Un élément optique adaptatif permet de recevoir la lumière guidée provenant de l'objet et de compenser la position du champ dépendant des aberrations optiques et des dispositifs optiques sont situés le long d'au moins une partie de la voie de lumière de manière à conditionner et focaliser la lumière, à mesure qu'elle se déplace du miroir de guidage, le long de l'élément optique adaptatif, et jusqu'au plan d'image final.
EP05855789A 2005-01-27 2005-12-29 Microscope optique de scannage adaptatif Withdrawn EP1842095A4 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US64757205P 2005-01-27 2005-01-27
PCT/US2005/047287 WO2006081031A2 (fr) 2005-01-27 2005-12-29 Microscope optique de scannage adaptatif

Publications (2)

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EP1842095A2 true EP1842095A2 (fr) 2007-10-10
EP1842095A4 EP1842095A4 (fr) 2010-08-04

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EP05855789A Withdrawn EP1842095A4 (fr) 2005-01-27 2005-12-29 Microscope optique de scannage adaptatif

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EP (1) EP1842095A4 (fr)
JP (1) JP2008529082A (fr)
CN (1) CN100585449C (fr)
IL (1) IL183131A0 (fr)
WO (1) WO2006081031A2 (fr)

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Also Published As

Publication number Publication date
CN100585449C (zh) 2010-01-27
CN101116023A (zh) 2008-01-30
IL183131A0 (en) 2007-09-20
WO2006081031A3 (fr) 2007-03-01
EP1842095A4 (fr) 2010-08-04
WO2006081031A2 (fr) 2006-08-03
JP2008529082A (ja) 2008-07-31

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