WO2020153996A1 - Dynamic focus and zoom system for use wide-field, confocal and multiphoton microscopes - Google Patents

Dynamic focus and zoom system for use wide-field, confocal and multiphoton microscopes Download PDF

Info

Publication number
WO2020153996A1
WO2020153996A1 PCT/US2019/050976 US2019050976W WO2020153996A1 WO 2020153996 A1 WO2020153996 A1 WO 2020153996A1 US 2019050976 W US2019050976 W US 2019050976W WO 2020153996 A1 WO2020153996 A1 WO 2020153996A1
Authority
WO
WIPO (PCT)
Prior art keywords
mems mirror
fixed lens
mems
mirror
lens
Prior art date
Application number
PCT/US2019/050976
Other languages
English (en)
French (fr)
Inventor
Sarah LUKES
Original Assignee
Agile Focus Designs, LLC
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Priority claimed from US16/258,455 external-priority patent/US10466461B2/en
Application filed by Agile Focus Designs, LLC filed Critical Agile Focus Designs, LLC
Priority to JP2020513316A priority Critical patent/JP6978592B2/ja
Priority to KR1020207010628A priority patent/KR20210119278A/ko
Priority to CN201980005037.4A priority patent/CN111344620B/zh
Publication of WO2020153996A1 publication Critical patent/WO2020153996A1/en

Links

Classifications

    • GPHYSICS
    • G02OPTICS
    • G02BOPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00Microscopes
    • G02B21/24Base structure
    • G02B21/241Devices for focusing
    • GPHYSICS
    • G02OPTICS
    • G02BOPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B17/00Systems with reflecting surfaces, with or without refracting elements
    • G02B17/08Catadioptric systems
    • G02B17/0896Catadioptric systems with variable magnification or multiple imaging planes, including multispectral systems
    • GPHYSICS
    • G02OPTICS
    • G02BOPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00Microscopes
    • G02B21/02Objectives
    • G02B21/025Objectives with variable magnification
    • GPHYSICS
    • G02OPTICS
    • G02BOPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B26/00Optical devices or arrangements for the control of light using movable or deformable optical elements
    • G02B26/08Optical devices or arrangements for the control of light using movable or deformable optical elements for controlling the direction of light
    • G02B26/0816Optical devices or arrangements for the control of light using movable or deformable optical elements for controlling the direction of light by means of one or more reflecting elements
    • G02B26/0825Optical devices or arrangements for the control of light using movable or deformable optical elements for controlling the direction of light by means of one or more reflecting elements the reflecting element being a flexible sheet or membrane, e.g. for varying the focus
    • GPHYSICS
    • G02OPTICS
    • G02BOPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B26/00Optical devices or arrangements for the control of light using movable or deformable optical elements
    • G02B26/08Optical devices or arrangements for the control of light using movable or deformable optical elements for controlling the direction of light
    • G02B26/0816Optical devices or arrangements for the control of light using movable or deformable optical elements for controlling the direction of light by means of one or more reflecting elements
    • G02B26/0833Optical devices or arrangements for the control of light using movable or deformable optical elements for controlling the direction of light by means of one or more reflecting elements the reflecting element being a micromechanical device, e.g. a MEMS mirror, DMD
    • GPHYSICS
    • G02OPTICS
    • G02BOPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B15/00Optical objectives with means for varying the magnification
    • G02B15/14Optical objectives with means for varying the magnification by axial movement of one or more lenses or groups of lenses relative to the image plane for continuously varying the equivalent focal length of the objective

Definitions

  • the present invention relates generally to the field of microscope technology and, more specifically, to a dynamic focus and zoom system for use with wide-field, confocal and multiphoton microscopes.
  • variable-power/variable-focus/varifocus (these terms are used synonymously here) optical elements perform fast focusing like the human eye. They remain stationary as their focal length changes.
  • Three varifocus lenses allow both magnification and focus to be changed independently of one another, and they also allow for a greater range of focus than use of a single varifocus lens.
  • Varifocus lenses include both transmissive and reflective lenses (mirrors), which behave the same except for the direction in which the light travels after hitting their surfaces.
  • the present invention incorporates varifocus lenses into a device that can be used with or attached to a microscope to overcome the disadvantages of conventional microscopes.
  • Micro-electro-mechanical systems (MEMS) deformable mirrors are reflective lenses with variable focal length. With electrostatic MEMS mirrors, voltage is applied directly to them for fine control of their surface shape. MEMS mirrors are commonly, but not exclusively, made of a photosensitive polymer, SU-8 2002, and a metal such as aluminum, gold or silver (which provides the reflective surface). In the context of the present invention, the MEMS mirrors may be made of silicon nitride in lieu of SU-8 2002, with a titanium dioxide or a metal/oxide reflective surface. As the radius of the curvature of the mirror decreases with increasing voltage, the location of the object (that is being viewed) changes. The capability for these types of MEMS mirrors to replace magnifiers or perform significant focusing has only become possible in approximately the last five years. These advancements came from improved microfabrication techniques and structural changes to the mirrors.
  • Disadvantages of traditional focusing mechanisms include agitation of the sample with translation of the stage (which can result in difficulty finding focus when observing solid particulates in liquids, damage to samples when they accidentally hit the objective lens, slow focus control (typically less than 100 Hz speed even with automated stages), inaccurate focusing, excessive photodamage from overexposure, early termination of experiments due to reaction completion prior to observation (due to the excessive time it takes to find a focal plane of interest), and the inability to observe biological phenomenon at small timescales due to speed-of-imaging limitations, such as observation of neurons propagating in real-time in living samples. Automated sample stages are limited to discrete focus step locations and traditional zoom systems to discrete magnification changes.
  • a resonant speed objective lens scanner can quickly translate the objective lens up and down.
  • Resonant objective lens scanners do not maintain optical resolution over the focusing range, operate only at a single (relatively fast) speed, and have a moving mass, which causes them to vibrate or shake the system as they resonate.
  • varifocus lenses including MEMS mirrors
  • MEMS mirrors have been used in connection with some confocal and multiphoton microscopes to achieve greater focus, they have not been used for optical zoom in microscopy, nor have they been incorporated into a small and portable unit that is user-friendly for the common biologist.
  • varifocus lenses are difficult to use for the average biologist; for example, most dynamic focusing mechanisms that are situated at the beginning of the optical train in a microscope require installation by an engineer. These are usually custom jobs with a single varifocus element done at elite universities for one experiment only, thereby precluding widespread use. Liquid lenses, which are another form of varifocus lens, do not maintain optical resolution over the focusing range, are not achromatic, and cannot effectively control attendant spherical aberration.
  • the present invention would allow a scientist to observe the entire cell membrane at both low and high magnification while keeping his or her hands free (through the use of voice activation or a foot step button).
  • the present invention maintains the resolution of the system over the focus range, allows for variable speed focusing, and allows the sample to remain stationary.
  • U.S. Patent No. 7864996 discloses a tissue attachment device in which the macroscopic imager and confocal imager are each individually presented to the tissue in a predefined alignment such that imaging locations of the confocal imager with respect to the tissue surface spatially correlate with the macroscopic image.
  • a turret rotates and switches between macroscopic and microscopic objective lenses, whereas in the present invention, magnification is changed by altering voltages across the stationary MEMS mirrors.
  • the present invention has no translation or moveable parts or motor to drive moveable parts, which constitutes a significant structural difference as compared to Hemmer and other similar microscopes. Furthermore, as opposed to switching between two separate instruments to switch from macroscopic to microscopic (confocal) imaging, the present invention has one stationary unit that changes magnification by altering voltages on MEMS mirrors.
  • U.S. Patent Nos. 8425037 (Uhlhorn et al, 2013) and 9492080 (Uhlhorn et al. , 2016) provide an optical coherence tomography (OCT) system that may be coupled to an operation microscope so that a surgeon can visualize ocular structures like the human crystalline lens and other ocular structures such as the cornea and/or vitreous while surgical instruments are in the field of view.
  • OCT optical coherence tomography
  • this particular invention does not attempt to solve the same problems as the present invention, is not structurally similar to the present invention, and does not have the same application as the present invention.
  • Both the OCT and spectrometer described in these patents have an interferometer.
  • An interferometer has two paths of light that recombine and interfere.
  • the present invention has one path of light going forward and one path returning. There is no recombination of the two paths.
  • the Ulhorn invention is not a MEMS device that operated under an applied voltage. Rather, the surgical microscope has a fixed focal length and is used to visually find a specific location in the eye, at which point the entire instrument is locked into place, and the imaging is done via OCT. The change in OCT focusing is accomplished by altering the optical pathlength of the interferometer.
  • U.S. Patent No. 9256009 discloses a microscope that incorporates a tunable acoustic gradient index of refraction (TAG) lens, which is a type of variable focal length lens, proximate to the microscope stage and a pulsed illuminator capable of illuminating the stage and synchronously pulsed with an operating frequency of the TAG lens.
  • TAG acoustic gradient index of refraction
  • One of the objects of this invention is to overcome the slow switching speeds of spatial light modulators.
  • the invention is directed primarily toward improving throughput for materials processing applications.
  • the lens is
  • Theriault s lens consists of two transparent windows and is filled with a refractive material (gas, solid, liquid, plasma, etc.). Light transmits through the lens and is not reflected as with a MEMS mirror. Theriault uses a single lens to accomplish both magnification and focusing, whereas the present invention uses three MEMS mirrors to perform these functions.
  • One drawback of Theriault’s single lens is that it requires a pulsed laser or camera and an iris for focusing, none of which is necessary in the present invention.
  • U.S. Patent No. 9602715 discloses an imaging system that is comprised of a variable magnification lens portion and a variable focal length lens (TAG lens) portion.
  • TAG lens variable focal length lens
  • This particular invention is intended to be used in a precision machine vision inspection or metrology system. It is not structurally or functionally similar to the present invention but does represent an improvement in magnification technology.
  • the Gladnick invention uses interchangeable lenses to change magnification states, similar to Hemmer (discussed above). Interchangeable lenses for magnification results in discrete magnification states.
  • the electronically controlled variable focus MEMS mirrors of the present invention allow for continuous magnification changes from one (minimum) magnification state to another (maximum).
  • his transmissive TAG lens operates at two discrete constant resonant frequencies that correspond with the two magnification states.
  • the MEMS mirrors of the present invention operate over a broad and continuous range of frequencies from DC to 1 kHz.
  • the present invention fills the back aperture of the objective lens for all magnifications and focusing.
  • Gladnick’s invention does not fill the back aperture of the objective lens, as an iris alters the size of the back aperture of the objective lens for each magni fication state of their design.
  • MEMS mirrors to provide dynamic focusing and zoom in wide-field, confocal and multiphoton microscopes.
  • the present invention there is no such system that is configured as a single, portable unit and designed for ease of use by a scientist in the lab.
  • the benefits of MEMS mirrors include the fact that they typically have significantly more focus range and higher bandwidth, they are less sensitive to temperature, and they are achromatic for fluorescence applications.
  • variable- power/variable-focus optical elements in the form of a single MEMS mirror
  • the details of the dynamic focus and zoom system provided herein have not heretofore been disclosed.
  • the 2014 paper did not address the use of three MEMS mirrors, aligned as described herein, to accomplish independent adjustment of focusing and zoom.
  • the basic structure of the MEMS mirror itself is discussed in the 2014 article.
  • the present invention is a dynamic focus and zoom system for use with wide- field, confocal and multiphoton microscopes comprising: a first MEMS mirror, a second MEMS mirror, and a third MEMS mirror situated within a housing and mounted onto an interior platform; a right-angle prism; an optical relay comprised of a first fixed lens and a second fixed lens; and a third fixed lens; wherein the first MEMS mirror, the second MEMS mirror, and the third MEMS mirror are elliptical in shape; wherein the right-angle prism, the first fixed lens, the second fixed lens, and the first MEMS mirror are aligned linearly along a longitudinal axis of the optical relay; wherein the second MEMS mirror, the third fixed lens, and the third MEMS mirror are linearly aligned with one another parallel to the longitudinal axis of the optical relay; wherein the right-angle prism is situated proximate to a first portal in the housing and at a forty-five degree angle relative to the first fixed lens; wherein the first MEMS mirror
  • the present invention is a dynamic focus and zoom system for use with wide-field, confocal and multiphoton microscopes comprising: a first MEMS mirror, a second MEMS mirror, and a third MEMS mirror situated within a housing and mounted onto an interior platform; a right-angle flat mirror; an optical relay comprised of a first fixed lens and a second fixed lens; and a third fixed lens; wherein the first MEMS mirror, the second MEMS mirror, and the third MEMS mirror are elliptical in shape; wherein the right-angle flat mirror, the fixed lens, the second fixed lens, and the first MEMS mirror are aligned linearly along a longitudinal axis of the optical relay; wherein the second MEMS mirror, the third fixed lens, and the third MEMS mirror are linearly aligned with one another parallel to the longitudinal axis of the optical relay; wherein the right-angle flat mirror is situated proximate to a first portal in the housing and at a forty-five degree angle relative to the first fixed lens; wherein the first MEMS mirror,
  • the first and second MEMS mirrors each has a minor axis with a diameter, and wherein the diameter of the minor axis of the second MEMS mirror is the same as the diameter of the minor axis of the first MEMS mirror.
  • the first and third MEMS mirrors each has a minor axis with a diameter, and the diameter of the minor axis of the third MEMS mirror is at least twice the diameter of the minor axis of the first MEMS mirror.
  • the first MEMS mirror has a minor axis with a diameter
  • the system is used in connection with an objective lens having a back aperture with a diameter
  • the diameter of the minor axis of the first MEMS mirror is the same as the diameter of the back aperture of the objective lens.
  • the first fixed lens, the second fixed lens, and the third fixed lens are preferably achromatic.
  • the right-angle prism is preferably a dielectric reflective prism.
  • Figure 1 is a first perspective view of a first embodiment of the present invention.
  • Figure 2 is a second perspective view of a first embodiment of the present invention.
  • FIG 3 is the same view as that shown in Figure 1 except with the top cover of the housing removed.
  • Figure 4 is the same view as that shown in Figure 3 except with the printed circuit boards removed.
  • FIG 5 is the same view as that shown in Figure 4 except with the spacers and mount plates removed.
  • FIG. 5A is the same view as that shown in Figure 5 except with the side walls of the housing and universal serial buses (USBs) removed.
  • USBs universal serial buses
  • Figure 5B is a bottom perspective view of the first embodiment of the present invention.
  • Figure 6 is a top view of the embodiment shown in Figure 5A.
  • Figure 7 is a third perspective view of the same embodiment shown in Figures 1 and 2.
  • Figure 8 is a first perspective view of a second embodiment of the present invention in which the objective lens is attached directly to the housing.
  • Figure 9 is a second perspecti ve view of the embodiment shown in Figure 9 in which the objective lens turret is attached directly to the housing.
  • Figure 10 is a diagram of the optical layout of the first and second embodiments of the present invention.
  • Figure 1 1 is a diagram of the optical train of the first and second embodiments of the present invention.
  • Figure 12 is a first top perspective view of a third embodiment of the present invention.
  • Figure 13 is a bottom perspective view of a third embodiment of the present invention.
  • Figure 14 is the same view as that shown in Figure 12 except with the top cover of the housing removed.
  • Figure 15 is a second top perspective view of a third embodiment of the present invention except with the top cover of the housing, printed circuit board and USB removed.
  • Figure 16 is the same view as that shown in Figure 15 except with the spacers and mount plate removed.
  • Figure 17 is the same view as that shown in Figure 16 except with the side walls of the housing removed.
  • Figure 18 is a top view of a third embodiment of the present invention.
  • Figure 19 is a diagram of the optical layout of a third embodiment of the present invention.
  • Figure 20 is a diagram of the optical train of a third embodiment of the present invention.
  • the present invention is comprised of a mechanical enclosure or housing that easily inserts into or attaches to existing widefield, confocal or multiphoton microscopes at an infinite conjugate image plane.
  • the invention is specifically designed to be situated between the objective lens turret and the objective lens, but it could also be placed near the eyepiece and function equivalently.
  • three electronically controlled variable focus lenses and one fixed lens perform significant defocusing and zoom.
  • the inventor determined that one MEMS mirror allows focusing but no zoom, two MEMS mirrors allow focusing and zoom but are dependent on each other (focusing cannot be changed without changing the zoom/magnification properties and vice versa), and three mirrors allow independent focusing and zoom (that is, either can be changed without affecting the other).
  • the present invention allows for a change in magnification of the microscope without turning the microscope objective lens turret to switch objectives and the sample remains stationary and un agitated.
  • the invention alters the focus position within the sample, the back focal plane of the objective lens remains filled.
  • the numerical aperture or resolution of the system remains constant, unlike other fast focusing technologies available.
  • the invention alters defocus independently of zoom.
  • the invention can alter optical (as opposed to digital that does not improve resolution) magnification independently of focus.
  • FIG. 1 is a first perspective view of a first embodiment of the present invention.
  • the present invention is comprised of a housing that forms an enclosure around the other components of the invention.
  • the housing has a top cover 2, side walls 3 and floor (not shown).
  • the top cover 2 may be attached, secured or connected to or joined with the side walls 3 in any manner; although shown here as attached to the side walls 3 with screws, the present invention is not limited to this particular manner of attaching the top cover 2 to the side walls 3.
  • the top cover 2 and side walls 3 may be integrally molded (part of the same part) or welded together.
  • the housing 1 may be made of any suitable, durable and rigid material (such as aluminum).
  • the top cover 2 is comprised of one or more air vents 5 for heat dissipation.
  • Figure 1 also shows the first USB 6.
  • the housing 1 is preferably configured to form a first alcove 7 sufficient in size and shape to allow the objective lens 8 to access a first side of a main window 9 and a second alcove 10 sufficient in size and shape to allow the objective lens turret or camera/eyepiece (not shown) to access a second side of the main window 9.
  • the top cover 2 and floor 4 (see Figure 5B) of the housing 1 are preferably flat, and the side walls are situated between the top cover 2 and floor 4 and perpendicular to each of them.
  • the two alcoves 7, 10 form an extension 1 1 of the housing 1 in which the main window 9 is situated.
  • the main window 9 extends across the entire front end of the extension 1 1 and wraps around both sides of the extension 1 1 to allow the objective lens 8 and objective lens turret (not shown) to access the main window 9 from opposite sides of the window (so that the objective lens and objective lens turret are longitudinally aligned with one another).
  • FIG. 2 is a second perspective view of a first embodiment of the present invention. This figure shows the second and third USBs 12, 13, respectively,
  • USBs 6, 12, 13 extend through cut-outs in the side walls 3 of the housing 1.
  • Each USB is situated proximate to a MEMS mirror inside of the housing ( ee Figure 5) and provides the drive signal for one of the three MEMS mirrors.
  • a printed circuit board 14 corresponds to each of the three MEMS mirrors ( see Figure 3), and the power for each printed circuit board is supplied through a hole in the housing below the USB (see, e.g., Figure 7).
  • FIG 3 is tire same view as that shown in Figure 1 except with the top cover of the housing removed.
  • This figure shows three printed circuit boards 14, each of which controls a MEMS mirror (see Figure 5). In future embodiments, it may be possible to control all three MEMS mirrors with a single printed circuit board.
  • Figure 4 is the same view as that shown in Figure 3 except with the printed circuit boards removed.
  • a plurality of spacers 15 are preferably situated on top of one or more mount boards 16 between the mount boards 16 and the printed circuit boards 14.
  • the mount boards 16 are situated beneath the printed circuit boards 14, and the purpose of the spacers 15 is to ensure that the printed circuit boards 14 do not come into electrical contact with the mount boards 16.
  • Holes 17 in the mount board 16 allow electrical wiring to pass through the mount board 16 to the printed circuit boards 14 from the USBs 13 below.
  • Figure 5 is the same view as that shown in Figure 4 except with the spacers and mount plates removed.
  • the invention comprises three MEMS mirrors 18, 19, 20, all of which are preferably circular.
  • part numbers 18, 19 and 20 are shown as small, square printed circuit boards with the circular mirrors adhered to them with tape or epoxy.
  • the invention preferably comprises an interior platform 21 that allows the various optical components to be aligned properly.
  • Figure 5 A is the same view as that shown in Figure 5 except with the side w'alls of the housing and USBs removed. Note that one or more holes 22 in the interior platform 21 allow the invention to be secured to an optical bench with screws.
  • a first prism 23 and a second prism 24, which is adjacent to the first prism 23, are situated just inside of the main window 9.
  • the main window 9 may be covered with glass or similar transparent material.
  • an optical relay comprised of a first fixed lens 25 and a second fixed lens 26.
  • the first and second fixed lenses are preferably slidably mounted within a slot 27 in the floor 4 of the housing 1 and fixed in place with set screws (not shown) so that the distance between them can be adjusted.
  • first beam splitter 28 Situated directly behind the second fixed lens 26 is a first beam splitter 28.
  • the first beam splitter 28 is situated so that the second prism 24, first fixed lens 25, second fixed lens 26, and first beam splitter 28 are all aligned linearly along the longitudinal axis of the optical relay.
  • a wave plate 29 is situated between the first beam splitter 28 and the first MEMS mirror 18.
  • the first and second MEMS mirrors 18, 19 are linearly aligned with one another at a ninety-degree angle to the longitudinal axis of the optical relay.
  • a second beam splitter 30 is situated in front of the second MEMS mirror 19, and a wave plate 29 is situated between the second beam splitter 30 and the second MEMS mirror 19.
  • the third MEMS mirror 20 is situated proximate to the main window 9.
  • a third fixed lens 31 is situated directly in front of the third MEMS mirror 20, and a wave plate 29 is situated directly in front of the third fixed lens 31.
  • the present invention was designed to be as small as possible so that it could be attached to existing microscopes without significant bulk.
  • the addition of the third fixed lens 31 made it possible to keep the length of the primary optical elements under four inches.
  • the third fixed lens 31 is positioned between the first prism 23 and the third prism 33.
  • the third fixed lens 31 is positioned between the second beam splitter 30 and the third beam splitter 32.
  • the third fixed lens 31 would have a different focal length depending on where it is placed, but this focal length may be calculated using the equations set forth below.
  • a third beam splitter 32 is situated directly in front of the wave plate 29 that is in front of the third fixed lens 31
  • a third prism 33 is situated directly in front of the third beam splitter 32.
  • the third MEMS mirror 20, third fixed lens 31 , wave plate 29, third beam splitter 32, and third prism 33 are all linearly aligned with one another at a ninety-degree angle to the longitudinal axis of the optical relay.
  • the third prism 33 abuts up against a center section of the optical relay (between the two fixed lenses 25, 26) and is also linearly aligned with the first prism 23 such that the linear alignment of these two prisms is parallel to the longitudinal axis of the optical relay.
  • the first, second and third beam splitter 28, 30 and 32 are all preferably polarizing beam splitters.
  • the distance between the first and second MEMS mirrors 18, 19 is less than the distance between the second and third MEMS mirrors 19, 20. This configuration is also shown in Figure 6.
  • FIG 6 is a top view of the embodiment shown in Figure 5A.
  • Microscopes can be used with transmissive illumination and reflective illumination (where light travels forwards through the system and reflects back through it to the camera/eyepiece for viewing).
  • the camera/eyepiece and the objective lens can be placed in a few different locations.
  • the camera/eyepiece (or objective lens turret) is placed at position“X” on Figure 6, and the objective lens (which provides a source of illumination for transmission) is situated at position“Y” ( see also Figure 1).
  • the camera/eyepiece may be placed at position“Z” with the objective Jens at position“Y”; in this configuration, the first prism 23 is removed.
  • the camera/eyepiece is at position“X,” and the objective lens is at position“Z”; in this configuration, the second prism 24 is removed.
  • the camera/eyepiece may provide a source of illumination for reflection.
  • the first MEMS mirror 18 is mapped directly to the back focal plane of the objective lens with a 1 : 1 correction ratio.
  • the third MEMS mirror 20 has a larger beam diameter than the other two MEMS mirrors 18, 19 to effectuate greater magnification.
  • Configuration A except that with the removal of the first prism 23, light exits the device at position“Z” on Figure 6.
  • the camera/eyepiece is perpendicular to the objective lens 8.
  • Configuration C the camera/eyepiece is also perpendicular to the objective lens 8, but the second prism 24 is removed rather than the first prism 23.
  • the path of light is the same as described above for Configuration A, except that light enters the device through the front of the main window 9 and hits the first fixed lens 25 of the optical relay.
  • Figure 7 is a third perspective view of the same embodiment shown in Figures 1 and 2. This figure provides a clearer view of the main window 9, through which the first 23 and second 24 prisms, as well as the first fixed lens 25, can be seen. Note that the diameter of the first prism 23 is larger than the diameter of the second prism 24 because the light beam is larger in diameter by the time it hits the first prism 23. Similarly, the diameter of the MEMS min or 20 is larger than that of the other two MEMS mirrors 1.8,
  • Figure 8 is a first perspective view of a second embodiment of the present invention in which the objective lens is attached directly to the housing
  • Figure 9 is a second perspective view of the embodiment shown in Figure 8 in which the objective lens turret is attached directly to the housing.
  • the device can be attached directly to an existing microscope.
  • the objective lens 8 screws into the housing at position“Y” ⁇ see Figure 6 for position designations) via a first threaded adapter 34 that is integral to the housing 1.
  • the objective lens turret (not shown) screws into the housing at position“X” via a second threaded adapter 35 that is also integral to the housing 1.
  • the main window 9 has been eliminated (because the objective lens and objective lens turret are screwed directly into the housing), and the extension 1 1 has been lengthened to accommodate the threaded adapters 34, 35, but in all other respects, the invention is as previously described.
  • Figure 10 is a diagram of the optical layout of the first and second embodiments of the present invention. Shown are one fixed lens F (reference number 31) and three varifocus lenses vJ, v2, and vi (reference numbers 20, 19 and 18, respectively). Fixed lens F and varifocus lens vl can be combined and are described as G. Varifocus lenses v2 and vi can be combined and described as H. Distance a represents the separation between fixed focal length lens F and varifocus lens vJ.
  • Distance b defines the separation between varifocus lens vl, and varifocus lens v2.
  • Distances b and c are positive.
  • Distance a can be positive as shown, where it is in front of varifocus lens vl.
  • Distance a can also be negative and behind varifocus lens vl. It is expected that the easiest and best place for F will be close to vl.
  • Distances a and c are assumed to be sufficiently small to consider the system a two-lens relay.
  • the equivalent focal lengths for G and H.fc and fy, respectively, are: where fr, ft, f z and fs are focal lengths for lenses F, vl, v2, and vi, respectively.
  • magnification of the sample observed at the second alcove 10 is:
  • a 6-mm-diameter for the first MEMS mirror 18 (vi) matches the diameter of the back aperture of most 0.8 NA objective lenses on a scale of 1 : 1 and acts as the aperture stop to the invention. This allows for easy correction of spherical and other aberration in die system by fine control of the MEMS mirror shape. It also ensures that the aperture stop of the entire system is the full aperture of the objective lens so that resolution is maintained over the dynamic focus range.
  • the 6 m MEMS aiirrors can have a focal length from 60 mm to infinity.
  • the space c between them is at least 20 ram due to the size of beam splitters and waveplates.
  • M -fc /36 2 or fc - 72 mm.
  • the third MEMS mirror 20 must have twice the diameter of MEMS mirrors 18, 19 for 2x magnification; therefore, its diameter is 12 mm.
  • MEMS mirror 20 has a focal length range of 200 m to infinity.
  • the value of 600 m for focal length/. / also allows the magnification of the system to be reduced by further decreasing/; ..
  • the distance between / and ft is less than 100 mm to keep the overall size of the unit relatively small.
  • the optical train shown in Figure 1 1 includes one 12 mm (vl) and two 6 mm (v2 and v3) MEMS mirrors; with reference to the above discussion of the figures, the larger MEMS mirror is the third MEMS mirror 20, and the two smaller MEMS mirrors are the first and second MEMS mirrors 18, 19.
  • Two relay lenses (the first and second fixed lenses 25, 26) image MEMS mirror 18 (v3) onto the back focal plane of the objective lens at 1 : 1 scale, which allows for easy correction of spherical aberration.
  • the back focal plane normally lies within the objective lens, and an optical relay allows for precise placement of a conjugate image plane there.
  • the placement of the two fixed lenses along this relay depends upon the microscope with which the device is being interfaced and the power of the two relay lenses.
  • Table 1 shows a few focus locations over a continuous 207- micron range in an aqueous sample at 0.8 NA. The table also shows a few
  • the present invention not only enables fine control over small amounts of defocus but also provides more than 200 pm focus range at 0.8 numerical aperture or more than 800 pm at 0.2 NA.
  • MEMS mirrors require around 100-150 volts to activate electronically.
  • the MEMS mirrors may be driven with bench-top high-voltage supplies.
  • a high-voltage boost converter is used to drive the MEMS mirrors from USB power.
  • the present invention is a vast improvement over the prior art in that it allows a lay person to perform fast focusing and zoom with a wide-field, confocal or multiphoton microscope without movement of the sample stage or objective lens.
  • Advantages of the present invention include, but are not limited to: (a) the sample remains stationary while focusing and zoom are all done optically within the instrument; (b) there is no translation of a large mass, so systemic vibration is minimized; (c) focus and zoom can be independently controlled; (d) relatively gross (in comparison with prior art) focusing and zoom is achieved in a small form factor: (e) the back aperture of the objective lens remains filled throughout the focal and zoom range, thus maintaining high resolution imaging; (f) variable and continuous focus speed is enabled; and (g) MEMS mirrors can operate at greater than 1 kHz speed for focusing and provide fine, continuous focus control.
  • Future embodiments of the present invention may include voice-activated control of focus and/or zoom, a foot step button for control of zoom so that the user can keep his or her hands free, software for controlling low-order and high-order spherical aberrations with concentric rings on the MEMS mirrors, software for controlling other aberration with variable lenses that have fine asymmetric shape control, and software for autofocus during time-lapse experiments.
  • Figures 12-20 depict a third embodiment of the present invention that is more compact than the first and second embodiments.
  • Figure 12 is a first top perspective view of a third embodiment of the present invention.
  • this embodiment comprises a housing 36 with a top cover 37, side walls 38 and floor 39.
  • the top cover 37 preferably comprises one or more air vents 40.
  • a single USB 41 extends through the top cover and provides the drive signal for the MEMS mirrors in this embodiment.
  • An objective lens 42 is attached to a first side wall 38 of the housing 36 via first threaded adapter 43 that is preferably integral to the housing 36; this attachment point is referred to herein as the first portal 58.
  • a hole 47 in the top of the housing allows a power cable to be connected to the printed circuit board 48. Other holes in the housing are for assembly purposes.
  • Figure 13 is a bottom perspective view of a third embodiment of the present invention.
  • the apparatus may be secured to an optical bench with screws 44.
  • the objective lens turret (not shown) is attached to a second side wall 38 of the housing 36 via a second threaded adapter 45 that is preferably integral to the housing 36. This attachment point is referred to herein as the second portal 59.
  • the slot 46 discussed further in connection with Figure 16, is also shown.
  • FIG 14 is the same view as that shown in Figure 12 except with the top cover of the housing removed.
  • the USB 41 is connected to a printed circuit board 48 within the housing 36.
  • the printed circuit board 48 controls all three MEMS mirrors 51 , 52, 56.
  • the objective lens 42 and threaded adapters 43, 45 are omitted from subsequent figures for clarity. Note that various threaded adapters may be used to connect either the objective lens 42 or the objective lens turret (not shown) to the housing 36 for the purpose of integrating with equipment from various manufacturers.
  • Figure 15 is a second top perspective view of the third embodiment shown with the printed circuit board and USB removed. As shown in this figure, a plurality of spacers 49 is situated between the printed circuit board 48 and the mount board 50.
  • FIG 16 is the same view as that shown in Figure 15 except with the spacers and mount plate removed.
  • the MEMS mirrors 51, 52, 56, prism/mirror 55, and third fixed lens 57 are all mounted on the base plate 60. which functions as an interior platform.
  • the prims/mirror 55 may be a right-angle flat mirror or a right-angle prism.
  • the slot 46 is also formed in the base plate/interior platform 60.
  • Figure 17 is the same view as that shown in Figure 16 except with the side walls of the housing removed.
  • FIG. 18 is a top view of the third embodiment of the present invention.
  • this embodiment comprises a first MEMS mirror 51 and a second MEMS mirror 52.
  • Each of these MEMS mirrors is preferably elliptical in shape, which eliminates the need for beam splitters and quarter wave plates to steer the light appropriately. The elimination of these components not only decreases light loss but also allows for a more compact design.
  • the use of el liptical MEMS mirrors also rids the system of astigmatic aberration, which occurs when using circular MEMS mirrors at an angle, thereby providing higher optical quality.
  • the astigmatism caused by circular MEMS mirrors and the ratio of axes used to overcome this astigmatism with elliptical MEMS mirrors is explained in the Ph.D. dissertation of the inventor, the contents of which are incorporated herein by reference. 2
  • the third embodiment of the present invention further comprises a slot 46 in the floor of the housing.
  • first and second fixed lenses 53, 54 form an optical relay and are slidably mounted within die slot 46 and fixed in place with set screws (not shown) so that the distance between them can be adjusted.
  • the end of the slot that is closest to the objective lens 42 is referred to herein as the‘proximal” end of the slot, and the end of the slot that is furthest from the objective lens is referred to herein as the“distal” end of the slot.
  • the first MEMS mirror 51 is aligned longitudinally with the first and second lenses 53, 54 at the distal end of the slot 46 so that a beam of light passing through the first and second lenses 53, 54 will hit the center of the first MEMS mirror 51.
  • the center points of the prism/mirror 55, first fixed lens 53, second fixed lens 54, and first MEMS mirror 51 are on the same horizontal plane.
  • the first MEMS mirror 51 is at an angle of forty-five (45) degrees relative to the second lens 54 so that the bea of light passing through the first and second lenses 52, 53 will hit the first MEMS mirror 51 at a forty-five (45)-degree angle.
  • the second MEMS mirror 52 is preferably the same size and shape as the first MEMS mirror 51. It is situated proximate to the first MEMS mirror 51 at an angle of ninety (90) degrees relative to the first MEMS mirror 51 and configured so that a beam of light hitting the first MEMS mirror 51 at an angle of forty-five (45) degrees will hit the center of the second MEMS mirror 52 at an angle of forty -five (45) degrees.
  • the prism/mirror 55 is a 6.25 mm dielectric reflective prism.
  • the first and second MEMS mirrors 51 , 52 are each 5 mm (minor axis) by 7.07 mm (major axis) diameter elliptical mirrors, and the minor axis of the first MEMS mirror 51 has the same diameter as the back aperture of the objective lens 42.
  • the third MEMS mirror 56 is a 10 mm (minor axis) by 14.14 (major axis) diameter elliptical mirror.
  • a reflective prism or mirror 55 that is situated inside the first portal 58 of the housing 36.
  • This prism/mirror 55 is aligned with the first and second lenses 53, 54 and the first MEMS mirror 51 so that they all lie on the same longitudinal axis (that is, the longitudinal axis of the optical relay), with the prism/mirror 55 being situated proximate to the proximal end of the slot 46.
  • the prism/mirror 55 is at a forty- five (45)-degree angle relative to the beam of light from the objective lens; therefore it is also at a forty-five (45)-degree angle relative to the first and second lenses 53, 54, which are oriented laterally on the slot 46.
  • the light ray is directed by the prism/miiTor 55 through the first and second lenses 53, 54, to the first MEMS mirror 51, to the second MEMS mirror 52, and then parallel to the slot 46 (or the longitudinal axis of the optical relay) until it hits a third MEMS mirror 56 that is situated adjacent to the prism/mirror 55 (preferably abutting the prism/mirror 55).
  • the center points of the second MEMS mirror 52, the third fixed lens 57, and third MEMS mirror 56 are on the same horizontal plane.
  • the third MEMS mirror 56 is at a forty-five (45)-degree angle to the beam of light coming from the second MEMS mirror 52, thereby directing the light through the second portal 59.
  • the light beam passes through a third fixed lens 57, the purpose of which is to shorten the optical path length and decrease the overall size of the invention.
  • the third fixed lens 57 is situated between the second and third MEMS minors 52, 56.
  • the third fixed lens 57 is achromatic with a diameter of 10mm and a focal length of 150 mm.
  • the third fixed lens 57 is shown as position-adjustable in this figure, it may or may not be movable in the final product.
  • Hitting the second MEMS mirror 52 at a forty-five (45)-degree angle the light is redirected at a ninety (90)-degree angle to the first MEMS mirror 51.
  • the first MEMS mirror 51 Once the light hits the first MEMS mirror 51, it is again redirected at a ninety (90)-degree angle and passes through the second lens 54 and then the first lens 53.
  • the light beam hits the prism/mirror 55, where it is redirected at a ninety (90)-degree angle and passes through the first portal 58 in the housing, through the objective lens, and to the sample, where it is reflected back through the objective lens, to the prism/mirror, etc.
  • the light retraces its initial path and is directed out through the second portal 59 in the housing, which, in this configuration, is the same portal through which the light entered the present invention before hitting the third MEMS minor 56.
  • the third MEMS mirror 56 is at least two times larger than the first MEMS mirror 51 for a 2x magnification change.
  • the third MEMS mirror 56 must be at least 1 .5 larger than the first MEMS mirror 51.
  • the third MEMS mirror 56 must be at least three times larger than the first MEMS mirror 51.
  • Figure 19 is a diagram of the optical layout of the third embodiment of the present invention. Shown are one fixed lens F (reference number 57) and three varifocus lenses vJ, v2, and v3 (reference numbers 56. 52, 51, respectively). Fixed lens and varifocus lens v/ can be combined and are described as G. Varifocus lenses v2 and v3 can be combined and described as H. Distance a represents the separation between fixed focal length lens and varifocus lens vl. Distance b defines the separation between fixed focal length lens F, and varifocus lens v2. Distances b and c are positive. Distance a can be positive as shown, where the fixed focal length lens is behind varifocus lens vl.
  • Distance a can also be negative and in front of varifocus lens vl. It is expected that the easiest and best place for F will be close to vl. Distances a and c are assumed to be sufficiently small to consider the syste a two-lens relay.
  • the equivalent focal lengths for G and H, fc and fa, respectively, are: where fa fa, fa, and fa are focal lengths for lenses F, vl, v2, and vi, respectively.
  • M - f r-
  • a 5-mm-diameter for the first MEMS mirror 51 matches the diameter of the back aperture of high NA air objective lenses on a scale of 1 : 1 and acts as the aperture stop to the invention. This allows for easy correction of spherical and other aberration in the system by fine control of the MEMS mirror shape.
  • the 5 mm MEMS mirrors can have a focal length from 60 mm to infinity.
  • distance b is limited to 190 mm to keep the invention small for ease of insertion and use between the objective lens turret and objective lenses in table top microscopes.
  • the third MEMS mirror 56 must have a minimum of twice the diameter (minor axis) of the first MEMS mirror 51 for 2x magnification; therefore, its diameter is 10 mm.
  • MEMS mirror 56 has a focal length range of 200 mm to infinity.
  • Figure 20 is a diagram of the optical train of the third embodiment of the present invention.
  • the optical train shown in Figure 20 includes one 10 m (v7) and two 5 mm (v2 and vJ) MEMS mirrors; with reference to the above discussion of the figures, the larger MEMS mirror is the third MEMS mirror 56, and the two smaller MEMS mirrors are the first and second MEMS mirrors 51 , 52.
  • Two relay lenses (the first and second fixed lenses 53, 54), which are not shown in this figure, image MEMS mirror 56 (v5) onto the back focal plane of the objective lens at 1 :1 scale, which allows for easy correction of spherical aberration.
  • the back focal plane normally lies within the objective lens, and an optical relay allows for precise placement of a conjugate image plane there.
  • the placement of the two relay lenses along this relay depends upon the microscope objective lens with which the device is being interfaced, as well as the focal length of each relay lens.
  • f F is the third fixed lens 57.
  • Table 2 shows a few focal lengths for the invention at lx magnification over a continuous range from -1367.8 mm to 109.1 mm in air without an attached objective lens. It also shows a few focal lengths for the invention at 2x magnification over a continuous range from -194.8 m to 98.5 mm in air without an attached objective lens.
  • Infinite-conjugate, off-the-shelf objective lenses are very precisely designed to minimize aberration for incoming light rays at infinite-conjugate.
  • the invention minimizes unwanted aberrations by keeping its own focal length centered about infinite-conjugate by having a negative to positive range.
  • the table shows the capability of the invention to maintain a focal distance of 200 n while changing magnification over a continuous range of lx-2x. If the invention is used with a 20x objective lens, then magnification of the imaged sample would range from 20x-40x.

Landscapes

  • Physics & Mathematics (AREA)
  • General Physics & Mathematics (AREA)
  • Optics & Photonics (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Spectroscopy & Molecular Physics (AREA)
  • Microscoopes, Condenser (AREA)
PCT/US2019/050976 2019-01-25 2019-09-13 Dynamic focus and zoom system for use wide-field, confocal and multiphoton microscopes WO2020153996A1 (en)

Priority Applications (3)

Application Number Priority Date Filing Date Title
JP2020513316A JP6978592B2 (ja) 2019-01-25 2019-09-13 広領域の共焦点及び多光子顕微鏡で用いる動的フォーカス・ズームシステム
KR1020207010628A KR20210119278A (ko) 2019-01-25 2019-09-13 광시야, 공초점 및 다광자 현미경과 함께 사용하기 위한 동적 초점 및 줌 시스템
CN201980005037.4A CN111344620B (zh) 2019-01-25 2019-09-13 用于宽场、共焦和多光子显微镜的动态聚焦和变焦***

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US16/258,455 2019-01-25
US16/258,455 US10466461B2 (en) 2017-09-15 2019-01-25 Dynamic focus and zoom system for use with wide-field, confocal and multiphoton microscopes

Publications (1)

Publication Number Publication Date
WO2020153996A1 true WO2020153996A1 (en) 2020-07-30

Family

ID=71736373

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2019/050976 WO2020153996A1 (en) 2019-01-25 2019-09-13 Dynamic focus and zoom system for use wide-field, confocal and multiphoton microscopes

Country Status (3)

Country Link
JP (1) JP6978592B2 (ja)
KR (1) KR20210119278A (ja)
WO (1) WO2020153996A1 (ja)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE102020212209A1 (de) 2020-09-28 2022-03-31 Robert Bosch Gesellschaft mit beschränkter Haftung Optikvorrichtung und Datenbrillensystem

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7002739B2 (en) * 2003-08-11 2006-02-21 Ohkura Industry Co. Device for generating line-shaped light beam and laser microscope
US20170199362A1 (en) * 2014-05-28 2017-07-13 Carl Zeiss Microscopy Gmbh Functionally integrated laser scanning microscope

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7057826B2 (en) * 2004-03-22 2006-06-06 Angstrom Inc. Small and fast zoom system
US7215882B2 (en) * 2004-07-21 2007-05-08 Angatrom, Inc. High-speed automatic focusing system
US7593156B2 (en) * 2005-08-26 2009-09-22 Leica Microsystems (Schweiz) Ag Microscope with micro-mirrors for optional deflection and/or beam splitting
DE102008041819A1 (de) * 2008-09-04 2010-03-11 Leica Microsystems (Schweiz) Ag Optisches Abbildungssystem

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7002739B2 (en) * 2003-08-11 2006-02-21 Ohkura Industry Co. Device for generating line-shaped light beam and laser microscope
US20170199362A1 (en) * 2014-05-28 2017-07-13 Carl Zeiss Microscopy Gmbh Functionally integrated laser scanning microscope

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE102020212209A1 (de) 2020-09-28 2022-03-31 Robert Bosch Gesellschaft mit beschränkter Haftung Optikvorrichtung und Datenbrillensystem

Also Published As

Publication number Publication date
JP6978592B2 (ja) 2021-12-08
JP2021513668A (ja) 2021-05-27
KR20210119278A (ko) 2021-10-05

Similar Documents

Publication Publication Date Title
US10168519B2 (en) Light sheet generator
JP5923783B2 (ja) 照明−および観察装置
US5282085A (en) Stereoscopic microscope including a field-magnifying lens in front of the objective lens
US11327288B2 (en) Method for generating an overview image using a large aperture objective
EP3682285B1 (en) Dynamic focus and zoom system for use with wide-field, confocal and multiphoton microscopes
JP2010061141A (ja) 顕微鏡カメラ用ビデオアダプタ
US10466461B2 (en) Dynamic focus and zoom system for use with wide-field, confocal and multiphoton microscopes
JP3534733B2 (ja) 固定高倍率切換型顕微鏡
US10842374B2 (en) Operating microscope
WO2020153996A1 (en) Dynamic focus and zoom system for use wide-field, confocal and multiphoton microscopes
US11327284B2 (en) Microscope system
JP2000284184A (ja) 平行系実体顕微鏡及び対物レンズ
US8529064B2 (en) Attachment module for a microscope for observing the fundus of the eye
CN111344620B (zh) 用于宽场、共焦和多光子显微镜的动态聚焦和变焦***
JPH11218681A (ja) 高解像マクロスコープ
JP2019076329A (ja) 前置レンズ装置及び眼科用顕微鏡
JP2003215472A (ja) 距離調節用光学拡大装置
JPH05107447A (ja) 双眼実体顕微鏡
JP2007256414A (ja) 顕微鏡装置
Haynes et al. The mechanical construction of the microscope
JPH05127070A (ja) 双眼実体顕微鏡
JP2009069295A (ja) ズーム顕微鏡

Legal Events

Date Code Title Description
ENP Entry into the national phase

Ref document number: 2020513316

Country of ref document: JP

Kind code of ref document: A

121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 19911444

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

32PN Ep: public notification in the ep bulletin as address of the adressee cannot be established

Free format text: NOTING OF LOSS OF RIGHTS PURSUANT TO RULE 112(1) EPC (EPO FORM 1205N DATED 15/09/2021)

122 Ep: pct application non-entry in european phase

Ref document number: 19911444

Country of ref document: EP

Kind code of ref document: A1