US20020068020A1 - Rapidly changing dichroic beamsplitter - Google Patents
Rapidly changing dichroic beamsplitter Download PDFInfo
- Publication number
- US20020068020A1 US20020068020A1 US09/988,880 US98888001A US2002068020A1 US 20020068020 A1 US20020068020 A1 US 20020068020A1 US 98888001 A US98888001 A US 98888001A US 2002068020 A1 US2002068020 A1 US 2002068020A1
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- United States
- Prior art keywords
- dichroic
- wheel
- microscope
- epi
- fluorescent
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- 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.)
- Abandoned
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- 230000005284 excitation Effects 0.000 claims abstract description 9
- 238000004458 analytical method Methods 0.000 description 2
- 230000003252 repetitive effect Effects 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 238000013459 approach Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 238000000295 emission spectrum Methods 0.000 description 1
- 238000000695 excitation spectrum Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
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Classifications
-
- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/16—Microscopes adapted for ultraviolet illumination ; Fluorescence microscopes
-
- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/06—Means for illuminating specimens
-
- 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
- G02B26/007—Optical devices or arrangements for the control of light using movable or deformable optical elements the movable or deformable optical element controlling the colour, i.e. a spectral characteristic, of the light
Definitions
- FIG. 3 is a top view showing the arrangement of a high speed dichroic beamsplitter wheel 13 constructed according to one embodiment of the invention.
- the wheel 13 includes a support plate 52 , on which a plurality of dichroic beamsplitter lenses 61 - 65 are arranged about a center axis 66 of the support plate 52 .
- Each of the lenses 61 - 65 have distinct filtering properties. In many cases, only two lenses are required, so it is possible to use plano lenses in the remaining three spots, or to leave the remaining three spots empty.
- the wheel is mounted at a 45 degree angle inside the microscope such that when a given dichroic is selected the opening holding that dichroic is rotated into the epi-fluorescent light path, and the selected dichroic beamsplitter is at 45 degree angle to the excitation light (FIG. 1, FIG. 4).
Abstract
An epi-fluorescent microscope is constructed with a quick changing set of filters. A wheel containing a plurality of dichroic beamsplitter mirrors mounted equidistance from a center axis of the wheel is mounted at a 45° angle within the lightpath and provides a beamsplitter function. Light from an excitation source is provided as monochromatic light, so that light from the excitation source is reflected by a selected one of the dichroic mirrors to the specimen, and Stokes shifted light from the specimen is transmitted through the dichroic mirror. By separately mounting the dichroic mirrors, the dichroic mirrors can be quickly moved and a quicker change of sensed fluorophores is achieved.
Description
- This application claims priority from U.S. Provisional Patent Application No. 60/249,626, filed Nov. 17, 2000, and is a continuation International Patent Application PCT/US01/42957. The entire contents of both of the aforementioned patent applications are incorporated herein by reference.
- This invention relates to epi-fluorescent microscopy, and more particularly to the rapid selection of filters for measuring fluorescence at different wavelengths or Stokes shifts.
- Current fluorescent microscope designs employ an incident light or epi-fluorescent design where a dichroic beam splitter (or chromatic mirror) mounted in a filter cube at a 45 degree angle to the excitation light path, is used to reflect shorter excitation wavelengths of light onto the specimen while passing longer emission wavelengths to the eyepieces or camera (FIG. 2).
- Many fluorescence applications require two or more fluorescent labels to be present in the specimen. Each label has its own excitation and emission spectra, and thus requires different excitation and emission filters, as well as a different dichroic beamsplitter.
- To date two approaches to using multiple fluorophores have been employed:
- 1) Use dichroic beamsplitters that have multiple cutoff wavelengths. Thus a single dichroic can be used with multiple fluorophores. However because of bandwidth restrictions, total light throughput is reduced, thus creating longer exposure times when working with a camera. Longer exposure times translate into longer acquisition duty cycles which is problematic in paradigms requiring repetitive high-speed data acquisition (screening applications and applications using living cells).
- 2) Motorized filter cube changers. Several commercially available microscopes employ motorized filter cube changes. These allow the use of single dichroics for each fluorophore. However switching time is slow (1-2 seconds) which creates problems when using multiple fluorophores in paradigms requiring repetitive high-speed data acquisition (screening applications and applications using living cells).
- Disclosed below is a device for rapidly changing dichroic beamsplitters in epi-fluorescent microscopes. The device is a high speed wheel in which dichroic beamsplitters are mounted. The high speed dichroic changer is mounted in an epi-fluorescent microscope, and the changer is under computer control. Computer software can command the changer to rotate different dichroic beamsplitters into the epifluorescent lightpath so that the appropriate dichroic is in position when a particular fluorophore is imaged. The present invention provides a microscope system in which the device is commanded by the software to change dichroic beamsplitters (FIG. 1).
- FIG. 1 is a system schematic showing some key components of an analysis microscope constructed according to the present invention.
- FIG. 2 is a schematic drawing showing epi-fluorescent light paths occurring in the system of FIG. 1.
- FIG. 3 is a top view showing the arrangement of a high speed dichroic beamsplitter wheel constructed according the present invention.
- FIG. 4 is a schematic drawing showing control of a microscope according to one aspect of the invention.
- FIG. 1 is a system schematic showing some key components of an
analysis microscope 11 constructed according to one aspect of the invention. The microscope includes alight source 12, adichroic beamsplitter wheel 13, and an objective 14 which is aligned with aspecimen 15. light from thelight source 12 is reflected at thedichroic wheel 13, and passes through the objective 14. Light reflected or fluorescing from thespecimen 15 again passes through the objective 14, and that light which passes through thedichroic wheel 13 is received by thecamera 16. Still referring to FIG. 1, thedichroic wheel 13 is rotationally positioned by astepper motor 17, which is controlled bymotor controller 18. Themotor controller 18 is responsive to acomputer 19, and thecomputer 19 also receives images from thecamera 16. - FIG. 2 is a schematic drawing showing epi-fluorescent light paths. Light from the
light source 12 is reflected by thedichroic beamsplitter wheel 13 which is at a reflection angle which determines the lightpath. In the exemplary embodiment, a 45 degree angle is used as the reflection angle, although since standard dichroic cubes with the 45 degree angle are not used, it is likely that other reflection angles can be used. The return light, if it is able to pass thedichroic wheel 13, is received by the camera. This sequence generally corresponds to fluorescent microscope designs which employ a dichroic beam splitter or a chromatic mirror mounted in a filter cube at a 45 degree angle to the excitation light path. Significantly the filter cube is not required. - FIG. 3 is a top view showing the arrangement of a high speed
dichroic beamsplitter wheel 13 constructed according to one embodiment of the invention. Thewheel 13 includes asupport plate 52, on which a plurality of dichroic beamsplitter lenses 61-65 are arranged about a center axis 66 of thesupport plate 52. Each of the lenses 61-65 have distinct filtering properties. In many cases, only two lenses are required, so it is possible to use plano lenses in the remaining three spots, or to leave the remaining three spots empty. - The solution described here is to mount round 50 mm dichroic beamsplitters in a high-speed filter wheel (FIG. 3). Any size or shape dichroic beamsplitter that matched the optical path of the microscope or instrument would be acceptable.
- The wheel is capable of switching between adjacent dichroics in 50 msec. The wheel in the current implementation is driven by a DC stepper motor and is under computer control. Switching time is a function of wheel mass and motor speed. Any motor-wheel combination that allowed faster switching times would be acceptable. It is also conceivable that a galvanometer could drive a wheel at much higher speeds, or that a galvanometer could also move dichroic beamsplitters mounted on a spindle.
- The wheel is mounted at a 45 degree angle inside the microscope such that when a given dichroic is selected the opening holding that dichroic is rotated into the epi-fluorescent light path, and the selected dichroic beamsplitter is at 45 degree angle to the excitation light (FIG. 1, FIG. 4).
- This design this allows optimized throughput for each fluorophore while at the same time being able to switch the dichroics rapidly. Optimized throughput is advantageous as it allows for shorter camera exposure times, which decreases duty cycle as well as helps reduce photobleaching. Being able to rapidly switch the dichroic beamsplitter reduces the interval between image acquisitions, thus reducing duty cycle.
- FIG. 4 is a schematic drawing showing control for a microscope using a fast dichroic beamsplitter changer according to one aspect of the invention. The figure depicts a
loop 80, which represents one complete duty cycle with respect to a predetermined wavelength. Application software sends amove command 82 to a dichroic beamsplitter wheel controller such ascontroller 18 in FIG. 1. The controller firmware then interprets 83 the move command and rotates 83 the wheel (13, FIG. 1) to a correct position. Thecontroller 18 then signals 84 application software thatwheel 13 has finished moving. In response, the computer (19, FIG. 1) signals 85 the camera (16, FIG. 1) to acquire 86 an image. The image is then acquired 87. - This sequence repeated by executing a
loop 80 for every wavelength required.
Claims (1)
1. A microscope with apparatus for rapidly changing dichroic beamsplitters in epi_fluorescent microscopes, the microscope comprising:
a source of excitation light;
a high speed wheel having a center axis bisecting a predetermined reflection angle within an epi-fluorescent lightpath of the microscope optics and rotable about the center axis driven by a changer;
a plurality of dichroic beamsplitter mirrors mounted to the wheel;
the wheel mounted in an epi-fluorescent microscope; and
a computer control, the changer responsive to the changer computer control, wherein computer software can command the changer to rotate different ones of the plurality of dichroic beamsplitters into the epi_fluorescent lightpath to position a selected one of the dichroic beamsplitter mirrors in the lightpath to image a particular fluorophore, thereby providing a microscope system in which the device is commanded by the software to change dichroic beamsplitters.
Priority Applications (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US09/988,880 US20020068020A1 (en) | 2000-11-17 | 2001-11-19 | Rapidly changing dichroic beamsplitter |
US10/825,134 US6927903B2 (en) | 2000-11-17 | 2004-04-16 | Rapidly changing dichroic beamsplitter |
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US24962600P | 2000-11-17 | 2000-11-17 | |
USPCT/US01/42957 | 2001-11-16 | ||
PCT/US2001/042957 WO2002041064A1 (en) | 2000-11-17 | 2001-11-16 | Rapidly changing dichroic beamsplitter |
US09/988,880 US20020068020A1 (en) | 2000-11-17 | 2001-11-19 | Rapidly changing dichroic beamsplitter |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/US2001/042957 Continuation WO2002041064A1 (en) | 2000-11-17 | 2001-11-16 | Rapidly changing dichroic beamsplitter |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US10/825,134 Continuation US6927903B2 (en) | 2000-11-17 | 2004-04-16 | Rapidly changing dichroic beamsplitter |
Publications (1)
Publication Number | Publication Date |
---|---|
US20020068020A1 true US20020068020A1 (en) | 2002-06-06 |
Family
ID=22944311
Family Applications (3)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US09/988,889 Expired - Lifetime US6683735B2 (en) | 2000-11-17 | 2001-11-19 | Rapidly changing dichroic beamsplitter in epifluorescent microscopes |
US09/988,880 Abandoned US20020068020A1 (en) | 2000-11-17 | 2001-11-19 | Rapidly changing dichroic beamsplitter |
US10/825,134 Expired - Lifetime US6927903B2 (en) | 2000-11-17 | 2004-04-16 | Rapidly changing dichroic beamsplitter |
Family Applications Before (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US09/988,889 Expired - Lifetime US6683735B2 (en) | 2000-11-17 | 2001-11-19 | Rapidly changing dichroic beamsplitter in epifluorescent microscopes |
Family Applications After (1)
Application Number | Title | Priority Date | Filing Date |
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US10/825,134 Expired - Lifetime US6927903B2 (en) | 2000-11-17 | 2004-04-16 | Rapidly changing dichroic beamsplitter |
Country Status (3)
Country | Link |
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US (3) | US6683735B2 (en) |
AU (1) | AU2002230415A1 (en) |
WO (1) | WO2002041064A1 (en) |
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Cited By (6)
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Also Published As
Publication number | Publication date |
---|---|
WO2002041064A1 (en) | 2002-05-23 |
US20020085293A1 (en) | 2002-07-04 |
US6927903B2 (en) | 2005-08-09 |
AU2002230415A1 (en) | 2002-05-27 |
US20040196547A1 (en) | 2004-10-07 |
US6683735B2 (en) | 2004-01-27 |
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Owner name: UNIVERSAL IMAGING CORPORATION, PENNSYLVANIA Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:STUCKEY, JEFFREY A.;REEL/FRAME:012591/0035 Effective date: 20020408 |
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STCB | Information on status: application discontinuation |
Free format text: EXPRESSLY ABANDONED -- DURING EXAMINATION |