Classifications

• • G—PHYSICS
  • G02—OPTICS
  • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
  • G02B27/00—Optical systems or apparatus not provided for by any of the groups G02B1/00 - G02B26/00, G02B30/00
  • G02B27/10—Beam splitting or combining systems
  • G02B27/14—Beam splitting or combining systems operating by reflection only
  • G02B27/145—Beam splitting or combining systems operating by reflection only having sequential partially reflecting surfaces
• • G—PHYSICS
  • G02—OPTICS
  • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
  • G02B21/00—Microscopes
  • G02B21/0004—Microscopes specially adapted for specific applications
  • G02B21/002—Scanning microscopes
• • G—PHYSICS
  • G02—OPTICS
  • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
  • 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
  • G02B21/0032—Optical details of illumination, e.g. light-sources, pinholes, beam splitters, slits, fibers
• • 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
  • G02B27/00—Optical systems or apparatus not provided for by any of the groups G02B1/00 - G02B26/00, G02B30/00
  • G02B27/10—Beam splitting or combining systems
  • G02B27/1086—Beam splitting or combining systems operating by diffraction only

Definitions

• the light (excitation light) of two lasers L1, L2 with different wavelengths is coupled into a common beam path via a mirror SP and a beam splitter ST, which on the side S1 of a mirrored prism in the direction of an AOTF (Acousto-OpticalTunable Filter ) is reflected.
• AOTF Acoustic-OpticalTunable Filter
• the excitation light is introduced into the AOTF, with light diffracted in the first order being deflected for the wavelength set via the control frequency of the AOTF exactly in the direction of a pinhole PH with upstream and downstream pinhole optics PHO for setting the beam profiles, while other possible wavelengths are undeflected in zero-order cross the AOTF and do not reach the pinhole.
• the pinhole PH serves here as an excitation and detection pinhole at the same time.
• the excitation light is imaged in the direction of a microscopic beam path MI in the direction of a sample via scanning units SC1, SC2 and a scanning optics SCO.
• the light emitted by the sample consisting of parts of the
• Excitation light and wavelength-shifted fluorescence components pass through the light path in the opposite direction to the AOTF.
• the wavelength components of the excitation light reach the mirror side S1 of the prism PS again via first-order diffraction, while the fluorescence components reach the AOTF cross undeflected in zero order and thereby take an angle to the reflected excitation light.
• the tip between prism surfaces S1 and S2 is now arranged between the returning zeroth and first order rays, as a result of which the fluorescent light strikes side S2 and from there in the direction of a detection unit, here consisting for example of a line filter LF, a color divider NFT and two Detectors for different wavelengths, is reflected.
• a detection unit here consisting for example of a line filter LF, a color divider NFT and two Detectors for different wavelengths, is reflected.
• the AOTF Due to the low bandwidth of the AOTF of approx. 2 nm bandwidth for the excitation light, it acts as an extreme edge filter with clear advantages, for example, against dichroic filters with bandwidths greater than 10 nm. This is particularly important because the distance between the excitation wavelength and fluorescence wavelengths is less than 10 nm can be and a wavelength-dependent separation is still possible due to the arrangement according to the invention.
• the AOTF can be switched from the wavelength of the laser L1 to the wavelength of the laser L2 and in turn the excitation light can be separated from the fluorescent light.
• FIG. 2 shows a similar arrangement with only one scanner SC.
• a mirror S is provided, which deflects the excitation light in the direction of the AOTF analogously to FIG. 1, here the fluorescence light returning in zero order runs through the AOTF next to the mirror S and in this way in the direction of one here detection, not shown.
• the AOTF can serve solely as a separation unit between excitation light and fluorescent light, by the laser light entering the AOTF in the direction of the first order without an upstream element and the detection light leaving the AOTF at an angle to the excitation light and directly entering a detection unit, which only has an effect on the overall length, since the angle of four degrees, for example, is right turns out small and overlaps of the wavelength components should be avoided.
• a separating mirror can also be provided only for the fluorescent light.
• a further advantageous embodiment in the form of an unmirrored prism is provided, which by refraction the light of an excitation laser in the first
• the invention is particularly advantageously applicable in a laser scanning ' microscope with an AOTF.
• AOTF and AOM are advantageously provided in the laser beam path for coupling the laser radiation.
• Several laser lines L1-L3 such as UV / VIS or IR can be coupled in simultaneously with independently adjustable excitation power or individually.

Landscapes

• Physics & Mathematics (AREA)
• General Physics & Mathematics (AREA)
• Optics & Photonics (AREA)
• Chemical & Material Sciences (AREA)
• Analytical Chemistry (AREA)
• Microscoopes, Condenser (AREA)
• Diffracting Gratings Or Hologram Optical Elements (AREA)

Priority Applications (5)

Applications Claiming Priority (4)

Publications (2)

Family

ID=26050938

Family Applications (1)

Country Status (5)

Cited By (5)

Families Citing this family (15)

Family Cites Families (27)

• 1999
  • 1999-08-03 DE DE19936573A patent/DE19936573A1/de active Pending
  • 1999-12-22 JP JP2000589988A patent/JP4532745B2/ja not_active Expired - Fee Related
  • 1999-12-22 EP EP99964647A patent/EP1141763B2/de not_active Expired - Lifetime
  • 1999-12-22 WO PCT/EP1999/010262 patent/WO2000037985A2/de not_active Ceased
  • 1999-12-22 DE DE59912534T patent/DE59912534D1/de not_active Expired - Lifetime
  • 1999-12-22 EP EP05012775.2A patent/EP1591825B2/de not_active Expired - Lifetime
  • 1999-12-22 DE DE59915074T patent/DE59915074D1/de not_active Expired - Lifetime
  • 1999-12-22 US US09/857,205 patent/US7009763B1/en not_active Expired - Fee Related

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