US20080304146A1 - Microscope System for Fcs Measurements - Google Patents
Microscope System for Fcs Measurements Download PDFInfo
- Publication number
- US20080304146A1 US20080304146A1 US12/088,639 US8863906A US2008304146A1 US 20080304146 A1 US20080304146 A1 US 20080304146A1 US 8863906 A US8863906 A US 8863906A US 2008304146 A1 US2008304146 A1 US 2008304146A1
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- United States
- Prior art keywords
- target
- light
- illuminating
- microscope system
- light source
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- 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
- G02B21/0052—Optical details of the image generation
- G02B21/0076—Optical details of the image generation arrangements using fluorescence or luminescence
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N21/6456—Spatial resolved fluorescence measurements; Imaging
- G01N21/6458—Fluorescence microscopy
-
- 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
- G02B21/0052—Optical details of the image generation
- G02B21/0064—Optical details of the image generation multi-spectral or wavelength-selective arrangements, e.g. wavelength fan-out, chromatic profiling
Definitions
- the invention relates to a microscope system for Fluorescence Correlation Spectroscopy (FCS) measurements, and in particular, to a microscope system for conducting FCS measurements.
- FCS Fluorescence Correlation Spectroscopy
- European Patent EP 0 941 470 describes a fluorescence correlation spectroscopy module for a microscope.
- the FCS module can additionally be connected to a microscope of any desired design. Fluorescence correlation spectroscopy allows the investigation of molecular dynamic processes to be studied. For this purpose, the particles contained in solution are doped with fluorescent dyes, and these dyes are then excited by light of a particular wavelength. The excitation light coming from a laser is coupled into the module via a flange joint for an optical waveguide. In the FCS module known from prior art, it is difficult to align the FCS detection volume with the sample area, which is to be investigated.
- a microscope system for conducting FCS measurements includes an illuminating light source configured to emit an illuminating and a target light source configured to emit a target light for marking an FCS volume in a sample volume.
- the wavelength of the illuminating light differs from the wavelength of the target light.
- the system also include a plurality of optical elements configured to direct the illuminating light and the target light onto the sample volume.
- FIG. 1 shows a schematic illustration of a first embodiment of the invention
- FIG. 2 shows a schematic illustration of a second embodiment of the invention.
- the present invention is directed to creating a microscope system which can be used to reliably perform the alignment with the sample volume to be investigated. This can be achieved by a microscope system comprising the features described below.
- the microscope system for conducting Fluorescence Correlation Spectroscopy (FCS) measurements can be provided with a target light source for marking an FCS volume.
- the light of the target light source can be also directed onto the sample volume via the plurality of optical elements.
- the wavelength of the first light source preferably differs from the wavelength of the target light source.
- a combining element can be provided which combines the illuminating light of the first light source with the light of the target light source to form a common beam path.
- the light of the target light source preferably has a longer wavelength than the illuminating light of the first light source.
- the light of the target light source can have a wavelength that is in the region of red light.
- the light of the target light source can have a wavelength that is in the region of IR light.
- a camera is provided which registers the IR light and converts it into an image visible to the user.
- the target light source is preferably provided with a correcting optics in order to compensate chromatic aberrations due to the different wavelengths of the first light source and the target light source.
- the first light source and/or the target light source can include a laser.
- the microscope is provided with an optical fiber into which the illuminating light of the at least first light source and the light of the target light source can be coupled in order to achieve the collinearity of the illuminating light and the light of the target light source.
- the combining element can include a beam splitter.
- the combining element can include an AOTF, an AOBS or an AOM.
- FIG. 1 schematically describes a microscope system 1 for conducting FCS measurements.
- the microscope system 1 is provided with at least one first light source 3 which emits illuminating light which is directed onto a sample volume 5 or a sample.
- a target light source 7 for marking the FCS volume 5 is provided.
- the target light source 7 emits light 6 , which is also directed onto the FCS volume.
- the wavelength of the illuminating light 2 of the first light source 3 differs from the wavelength of the light 6 of the target light source 7 .
- the light 2 from the illuminating light source 3 and the light 6 from the target light source 7 are combined by a combining element 9 to form a common, collinear beam path.
- the combining element 9 can be designed to include a beam splitter.
- the combining element 9 can include an AOTF, an AOBS or an AOM.
- a correcting optics 10 is provided between the target light source 7 and the combining element 9 , in order to compensate chromatic aberrations due to the different wavelengths of the light 2 of the first light source 3 and the light 6 of the target light source 7 .
- the light 2 of the first light source 3 and the light 6 of the target light source 7 is directed onto the sample volume 5 or the volume via a plurality of optical elements 12 and a microscope optics 14 .
- the sample volume 5 or sample is preferably provided at least on an X-Y table 16 , in order thereby to change the sample volume with respect to the position of the illuminating light.
- the sample volume 5 is excited to fluoresce due to the illumination by the first light source 3 , so that the sample volume 5 emits a detection light 15 , which is also directed onto the detector 18 via the microscope optics 14 and the optical elements.
- the light 6 of the target light source 7 has a longer wavelength than the illuminating light 2 of the first light source 3 .
- the light 2 of the target light source 3 has a wavelength lying in the region of red light. The location of the light 6 of the target light source 7 on the sample volume 5 can therefore be observed directly and visually by a user 24 . If the light 6 of the target light source 7 lies in the wavelength region of IR light, a camera 22 is provided which produces an image for the user 24 , so that the latter can recognize the location of the illuminating light 25 in the sample volume 5 .
- FIG. 2 shows a further embodiment of the microscope system 1 .
- an optical fiber 30 Arranged downstream of the combining element 9 is an optical fiber 30 into which the illuminating light 2 of the at least first light source 3 and the light 6 of the target light source are coupled.
- the collinearity of the illuminating light is achieved by coupling the illuminating light 2 of the light 6 of the target light source 7 into the optical fiber 30 . This ensures that the light 6 of the target light source 7 and the illuminating light 2 of the at least first light source 3 impinge on a shared impingement location 25 in the sample volume 5 or in the sample.
- the optical fiber 30 can be provided with a coupling-in optics 31 and a coupling-out optics 32 .
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- Physics & Mathematics (AREA)
- General Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Optics & Photonics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Microscoopes, Condenser (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
A microscope system for conducting FCS measurements. The system includes an illuminating light source configured to emit an illuminating light at an illuminating wavelength. A target light source is provided and configured to emit a target light for marking an FCS volume in a sample volume at a target wavelength. The target wavelength differs from the illuminating wavelength. The system further includes a plurality of optical elements configured to direct the illuminating light and the target light onto the sample volume.
Description
- This is a U.S. national phase application under 35 U.S.C. §371 of International Patent Application No. PCT/EP2006/066861, filed Sep. 28, 2006, and claims benefit of German Patent Application No. 10 2005 046 510.2, filed Sep. 29, 2005, which is incorporated by reference herein. The International Application was published in German on Apr. 5, 2007 as WO 2007/036559 A1 under PCT Article 21(2).
- The invention relates to a microscope system for Fluorescence Correlation Spectroscopy (FCS) measurements, and in particular, to a microscope system for conducting FCS measurements.
- European Patent EP 0 941 470 describes a fluorescence correlation spectroscopy module for a microscope. The FCS module can additionally be connected to a microscope of any desired design. Fluorescence correlation spectroscopy allows the investigation of molecular dynamic processes to be studied. For this purpose, the particles contained in solution are doped with fluorescent dyes, and these dyes are then excited by light of a particular wavelength. The excitation light coming from a laser is coupled into the module via a flange joint for an optical waveguide. In the FCS module known from prior art, it is difficult to align the FCS detection volume with the sample area, which is to be investigated.
- In accordance with the present invention, a microscope system for conducting FCS measurements is provided. The microscope system includes an illuminating light source configured to emit an illuminating and a target light source configured to emit a target light for marking an FCS volume in a sample volume. The wavelength of the illuminating light differs from the wavelength of the target light. The system also include a plurality of optical elements configured to direct the illuminating light and the target light onto the sample volume.
- In the drawings, the subject matter of the invention is illustrated schematically, and will be described in the following with the aid of the figures, in which:
-
FIG. 1 shows a schematic illustration of a first embodiment of the invention; and -
FIG. 2 shows a schematic illustration of a second embodiment of the invention. - The present invention is directed to creating a microscope system which can be used to reliably perform the alignment with the sample volume to be investigated. This can be achieved by a microscope system comprising the features described below.
- In accordance with one embodiment of the invention the microscope system for conducting Fluorescence Correlation Spectroscopy (FCS) measurements can be provided with a target light source for marking an FCS volume. Here, the light of the target light source can be also directed onto the sample volume via the plurality of optical elements. The wavelength of the first light source preferably differs from the wavelength of the target light source.
- A combining element can be provided which combines the illuminating light of the first light source with the light of the target light source to form a common beam path. The light of the target light source preferably has a longer wavelength than the illuminating light of the first light source.
- In accordance with a further aspect of the present invention, the light of the target light source can have a wavelength that is in the region of red light. In the same way, the light of the target light source can have a wavelength that is in the region of IR light. In the case of IR light, a camera is provided which registers the IR light and converts it into an image visible to the user. Furthermore, the target light source is preferably provided with a correcting optics in order to compensate chromatic aberrations due to the different wavelengths of the first light source and the target light source.
- In accordance with yet a further feature of the one embodiment of the present invention, the first light source and/or the target light source can include a laser.
- In a further embodiment, the microscope is provided with an optical fiber into which the illuminating light of the at least first light source and the light of the target light source can be coupled in order to achieve the collinearity of the illuminating light and the light of the target light source. In this case, the combining element can include a beam splitter. Alternatively, the combining element can include an AOTF, an AOBS or an AOM.
- Further advantageous refinements of the invention can be found in the discussion below.
-
FIG. 1 schematically describes amicroscope system 1 for conducting FCS measurements. Themicroscope system 1 is provided with at least one first light source 3 which emits illuminating light which is directed onto a sample volume 5 or a sample. Additionally, a target light source 7 for marking the FCS volume 5 is provided. The target light source 7 emitslight 6, which is also directed onto the FCS volume. The wavelength of theilluminating light 2 of the first light source 3 differs from the wavelength of thelight 6 of the target light source 7. Thelight 2 from the illuminating light source 3 and thelight 6 from the target light source 7 are combined by a combiningelement 9 to form a common, collinear beam path. In this case, the combiningelement 9 can be designed to include a beam splitter. Optionally, the combiningelement 9 can include an AOTF, an AOBS or an AOM. A correctingoptics 10 is provided between the target light source 7 and the combiningelement 9, in order to compensate chromatic aberrations due to the different wavelengths of thelight 2 of the first light source 3 and thelight 6 of the target light source 7. Thelight 2 of the first light source 3 and thelight 6 of the target light source 7 is directed onto the sample volume 5 or the volume via a plurality ofoptical elements 12 and amicroscope optics 14. The sample volume 5 or sample is preferably provided at least on an X-Y table 16, in order thereby to change the sample volume with respect to the position of the illuminating light. The sample volume 5 is excited to fluoresce due to the illumination by the first light source 3, so that the sample volume 5 emits adetection light 15, which is also directed onto thedetector 18 via themicroscope optics 14 and the optical elements. Thelight 6 of the target light source 7 has a longer wavelength than theilluminating light 2 of the first light source 3. In a first embodiment, thelight 2 of the target light source 3 has a wavelength lying in the region of red light. The location of thelight 6 of the target light source 7 on the sample volume 5 can therefore be observed directly and visually by auser 24. If thelight 6 of the target light source 7 lies in the wavelength region of IR light, acamera 22 is provided which produces an image for theuser 24, so that the latter can recognize the location of theilluminating light 25 in the sample volume 5. -
FIG. 2 shows a further embodiment of themicroscope system 1. Arranged downstream of the combiningelement 9 is anoptical fiber 30 into which theilluminating light 2 of the at least first light source 3 and thelight 6 of the target light source are coupled. The collinearity of the illuminating light is achieved by coupling theilluminating light 2 of thelight 6 of the target light source 7 into theoptical fiber 30. This ensures that thelight 6 of the target light source 7 and theilluminating light 2 of the at least first light source 3 impinge on a sharedimpingement location 25 in the sample volume 5 or in the sample. Theoptical fiber 30 can be provided with a coupling-inoptics 31 and a coupling-outoptics 32.
Claims (20)
1-10. (canceled)
11. A microscope system for conducting FCS measurements comprising:
an illuminating light source configured to emit an illuminating light having an illuminating wavelength;
a target light source configured to emit a target light for marking an FCS volume in a sample volume and having a target wavelength, the target wavelength being different than the illuminating wavelength; and
a plurality of optical elements configured to direct the illuminating light and the target light onto the sample volume.
12. The microscope system of claim 11 , further comprising a combining element configured to combine the illuminating light and the target light to form a common beam path.
13. The microscope system of claim 11 , wherein the target wavelength is longer than the illuminating wavelength.
14. The microscope system of claim 13 , wherein target wavelength is in a region of red light.
15. The microscope system of claim 13 , wherein the target light wavelength is in a region of IR light, the system further comprising a camera configured to detect IR light and convert the IR light into an image visible to a user.
16. The microscope system of claim 11 , wherein the target light source includes correcting optics configured to compensate for chromatic aberrations resulting from a difference between the target wavelength and the illuminating wavelength.
17. The microscope system of claim 11 , wherein at least one of the illuminating light source and the target light source includes a laser.
18. The microscope system of claim 11 , further comprising an optical fiber configured to receive the illuminating light and the target light so as to achieve a collinearity of the illuminating light and target light.
19. The microscope system of claim 12 , wherein the combining element includes a beam splitter.
20. The microscope system of claim 12 , wherein the combining element includes at least one of an AOTF, an AOBS, and an AOM.
21. The microscope system of claim 12 , wherein the target light source includes correcting optics configured to compensate for chromatic aberrations resulting from a difference between the target wavelength and the illuminating wavelength.
22. The microscope system of claim 13 , wherein the target light source includes correcting optics configured to compensate for chromatic aberrations resulting from a difference between the target wavelength and the illuminating wavelength.
23. The microscope system of claim 15 , wherein the target light source includes correcting optics configured to compensate for chromatic aberrations resulting from a difference between the target wavelength and the illuminating wavelength.
24. The microscope system of claim 12 , wherein at least one of the illuminating light source and the target light source includes a laser.
25. The microscope system of claim 15 , wherein at least one of the illuminating light source and the target light source includes a laser.
26. The microscope system of claim 16 , wherein at least one of the illuminating light source and the target light source includes a laser.
27. The microscope system of claim 13 , further comprising an optical fiber configured to receive the illuminating light and the target light so as to achieve a collinearity of the illuminating light and target light.
28. The microscope system of claim 15 , further comprising an optical fiber configured to receive the illuminating light and the target light so as to achieve a collinearity of the illuminating light and target light.
29. The microscope system of claim 16 , further comprising an optical fiber configured to receive the illuminating light and the target light so as to achieve a collinearity of the illuminating light and target light.
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DE102005046510.2A DE102005046510B4 (en) | 2005-09-29 | 2005-09-29 | Microscope system for FCS measurements |
DE102005046510.2 | 2005-09-29 | ||
PCT/EP2006/066861 WO2007036559A1 (en) | 2005-09-29 | 2006-09-28 | Microscope system for fcs measurements |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/EP2006/066861 A-371-Of-International WO2007036559A1 (en) | 2005-09-29 | 2006-09-28 | Microscope system for fcs measurements |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US13/210,554 Continuation US8300310B2 (en) | 2005-09-29 | 2011-08-16 | Method for FCS measurements |
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US20080304146A1 true US20080304146A1 (en) | 2008-12-11 |
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Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
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US12/088,639 Abandoned US20080304146A1 (en) | 2005-09-29 | 2006-09-28 | Microscope System for Fcs Measurements |
US13/210,554 Active US8300310B2 (en) | 2005-09-29 | 2011-08-16 | Method for FCS measurements |
Family Applications After (1)
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US13/210,554 Active US8300310B2 (en) | 2005-09-29 | 2011-08-16 | Method for FCS measurements |
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US (2) | US20080304146A1 (en) |
DE (1) | DE102005046510B4 (en) |
WO (1) | WO2007036559A1 (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US8259383B2 (en) | 2007-06-15 | 2012-09-04 | Leica Microsystems Cms Gmbh | Beam combiner and a light source with such a beam combiner |
Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20020121610A1 (en) * | 1996-11-29 | 2002-09-05 | Michael Tewes | Fluorescence correlation spectroscopy module for a microscope |
US20030071227A1 (en) * | 2001-10-16 | 2003-04-17 | Ralf Wolleschensky | Method for the optical acquisition of characteristic sizes of an illuminated sample |
US6693742B1 (en) * | 1999-10-26 | 2004-02-17 | Carl Zeiss Jena Gmbh | Arrangement for illumination with a plurality of wavelengths in a microscope |
US20040257562A1 (en) * | 2003-06-17 | 2004-12-23 | Leica Microsystems Heidelberg Gmbh | Method for measuring fluorescence correlations in the presence of slow signal fluctuations |
US20070109536A1 (en) * | 2003-06-20 | 2007-05-17 | The Regents Of The University Of California | Modulated excitation fluorescense analysis |
Family Cites Families (7)
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US4734570A (en) | 1985-11-20 | 1988-03-29 | Olympus Optical Co., Ltd. | Active focus detecting device with infrared source |
JPH06160724A (en) * | 1992-11-17 | 1994-06-07 | Nikon Corp | Flash photolysis microscope |
JP3917731B2 (en) | 1996-11-21 | 2007-05-23 | オリンパス株式会社 | Laser scanning microscope |
DE19702753C2 (en) | 1997-01-27 | 2003-04-10 | Zeiss Carl Jena Gmbh | Laser Scanning Microscope |
US7196843B2 (en) * | 2002-03-27 | 2007-03-27 | Olympus Optical Co., Ltd. | Confocal microscope apparatus |
DE502004009281D1 (en) * | 2003-09-25 | 2009-05-14 | Leica Microsystems | MICROSCOPE WITH EVANESCENT SPECIMEN LIGHTING |
DE10356826B4 (en) | 2003-12-05 | 2021-12-02 | Leica Microsystems Cms Gmbh | Scanning microscope |
-
2005
- 2005-09-29 DE DE102005046510.2A patent/DE102005046510B4/en not_active Expired - Fee Related
-
2006
- 2006-09-28 WO PCT/EP2006/066861 patent/WO2007036559A1/en active Application Filing
- 2006-09-28 US US12/088,639 patent/US20080304146A1/en not_active Abandoned
-
2011
- 2011-08-16 US US13/210,554 patent/US8300310B2/en active Active
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20020121610A1 (en) * | 1996-11-29 | 2002-09-05 | Michael Tewes | Fluorescence correlation spectroscopy module for a microscope |
US6693742B1 (en) * | 1999-10-26 | 2004-02-17 | Carl Zeiss Jena Gmbh | Arrangement for illumination with a plurality of wavelengths in a microscope |
US20030071227A1 (en) * | 2001-10-16 | 2003-04-17 | Ralf Wolleschensky | Method for the optical acquisition of characteristic sizes of an illuminated sample |
US20040257562A1 (en) * | 2003-06-17 | 2004-12-23 | Leica Microsystems Heidelberg Gmbh | Method for measuring fluorescence correlations in the presence of slow signal fluctuations |
US20070109536A1 (en) * | 2003-06-20 | 2007-05-17 | The Regents Of The University Of California | Modulated excitation fluorescense analysis |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US8259383B2 (en) | 2007-06-15 | 2012-09-04 | Leica Microsystems Cms Gmbh | Beam combiner and a light source with such a beam combiner |
Also Published As
Publication number | Publication date |
---|---|
US8300310B2 (en) | 2012-10-30 |
DE102005046510B4 (en) | 2022-02-17 |
DE102005046510A1 (en) | 2007-04-05 |
US20110299156A1 (en) | 2011-12-08 |
WO2007036559A1 (en) | 2007-04-05 |
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AS | Assignment |
Owner name: LEICA MICROSYSTEMS CMS GMBH, GERMANY Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:KNEBEL, WERNER;REEL/FRAME:021422/0067 Effective date: 20080402 |
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STCB | Information on status: application discontinuation |
Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION |