EP2181350A2 - Procédé et dispositif d'éclairage d'un échantillon dans un microscope laser - Google Patents

Procédé et dispositif d'éclairage d'un échantillon dans un microscope laser

Info

Publication number
EP2181350A2
EP2181350A2 EP08787083A EP08787083A EP2181350A2 EP 2181350 A2 EP2181350 A2 EP 2181350A2 EP 08787083 A EP08787083 A EP 08787083A EP 08787083 A EP08787083 A EP 08787083A EP 2181350 A2 EP2181350 A2 EP 2181350A2
Authority
EP
European Patent Office
Prior art keywords
laser light
light source
laser
amplifier
source
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
EP08787083A
Other languages
German (de)
English (en)
Inventor
Volker Seyfried
Hilmar Gugel
Carsten L. Thomsen
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Leica Microsystems CMS GmbH
Original Assignee
Leica Microsystems CMS GmbH
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
Application filed by Leica Microsystems CMS GmbH filed Critical Leica Microsystems CMS GmbH
Publication of EP2181350A2 publication Critical patent/EP2181350A2/fr
Ceased legal-status Critical Current

Links

Classifications

    • 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

Definitions

  • the invention relates to a method for illuminating or irradiating an object, a sample or the like for the purpose of image acquisition or analysis, in particular for use in a laser microscope, preferably in a confocal microscope, with a laser light source emitting the illumination light, wherein the laser light directly or via a fiber is coupled into an illumination beam path. Furthermore, the invention relates to a corresponding device.
  • the invention generally relates to a method and a device for illuminating or irradiating an object, a sample or the like, which in concrete terms may be the application in confocal microscopy.
  • Confocal microscopes with conventional gas, solid-state, semiconductor or fiber lasers, in particular with white-light lasers are known, for example, from DE 101 15 589 A1 or from DE 101 15 509 A1.
  • it is already known to produce a so-called supercontinuum in white-light lasers namely with the aid of a photonic glass fiber, a photonic bandgap fiber, a tapered fiber, a Holy fiber, a doped fiber, etc.
  • each laser ages in operation, for example by gas degradation.
  • the optics, the resonator, active media, etc. age due to thermal stress.
  • the photonic glass fiber degenerates, which is manifested by a gradual decrease in the output power over the operating period.
  • the laser illumination light is only used for a fraction of the time while the laser is constantly operating is on or remains on. So you need the laser light regularly only for the actual image capture and not in the usually long pauses between shots.
  • the present invention is therefore based on the object, a method and an apparatus for illuminating or irradiating an object, a sample or the like. indicate for the purpose of image acquisition or analysis, according to which in particular the components which are sensitive with respect to the laser lifetime are spared in particular when the laser is to be ready for operation, but the laser light is temporarily not required.
  • the generic method is characterized in that the laser light source on a
  • the generic device is achieved in that the laser light source in response to a trigger signal immediately before the actual need, for example, immediately before the image acquisition, is switched on very quickly.
  • the laser light source is switched on extremely quickly in response to a trigger signal. So that is guaranteed ⁇ ensure that the laser is actually only switched on if, for example, receives the con ⁇ fokalmikroskop images. If the laser light is not needed, the laser light source can be switched off manually, or preferably automatically after a certain time delay.
  • the trigger signal used to turn on the laser light source can be triggered manually or automatically by hardware or software. If the device using the laser light source is a microscope, the triggering signal required for switching on is then directed to the laser light source when the user starts or starts the preparations for image acquisition before the actual image acquisition. Due to the trigger signal, the laser light source is turned on.
  • the switching on of the laser light source is preferably carried out in less than 1/10 second, so that the user does not perceive the switch-on or the time until trouble-free and stable operation of the laser light source as disturbing.
  • the laser light source is switched on via a pump source of the laser light source, which is preferably designed as a pump diode. This pump source is energized, whereby the laser or the laser light source is activated.
  • a control device with a control loop which regulates the current of the pump source as a function of the output power of the laser light source is used.
  • the control device comprises a photodiode serving for detecting the output power of the laser light source, preferably a so-called monitor photodiode. The signal detected there is evaluated and serves as a controlled variable for the control loop.
  • a permanently operated or energized seed laser with at least one downstream laser amplifier is provided in a further advantageous manner.
  • the laser light source comprises the permanently operated seed laser and the laser amplifier to be switched on as needed. Permanent energization of the seed laser achieves stable conditions in the generation of the laser radiation.
  • the rapid switching of the radiation takes place by switching on or energizing the pump source for the laser amplifier.
  • a trigger or a trigger signal indicates that laser light is required, as a result of which the pump source, which is advantageously a pump diode or pump diodes, is switched on for the amplifier unit.
  • a fast regulation of Output power for this interval operation is again generated by a Montitor photodiode.
  • the laser amplifier is constructed in two or more stages, namely, the laser amplifier comprises at least one preamplifier and at least one power amplifier.
  • the laser amplifier or downstream laser amplifiers are activated by switching on or energizing the respective pump source while the seed laser is constantly operating.
  • the first stage of the amplifier unit is also operated permanently or energized.
  • a control device is provided with a control circuit, according to which the current of the pump source of the amplifier unit is controlled in dependence on the output power of the laser light source.
  • a photodiode preferably a monitor photodiode, is also provided here.
  • the control device can be the same control device that is required to control the pump light source of the seed laser.
  • a single control device can serve both to control the energization of the pump source of the seed laser and the pump sources of preamplifier and power amplifier.
  • the splitting of the laser light source or of the laser system is advantageous in particular because the components which are sensitive with regard to the laser lifetime are in the laser amplifiers or the laser amplifier are downstream. These critical components are operated according to the invention only if they are actually needed. Accordingly, synchronization of the switch-on processes or the current application of the respective pump sources takes place, for example synchronized with the scanning process of the confocal scanning microscope.
  • nonlinear optical elements such as frequency doublers or frequency multipliers, frequency mixers, frequency converters (OPOs, etc.) or spectrally broadening elements such as self-phase modulation or cross-phase modulation materials, photonic crystals, photonic crystal fibers, supercontinuum-producing fibers (photonic, microstructured, suitably doped or with natural negative group velocity dispersion) are arranged.
  • nonlinear optical elements such as frequency doublers or frequency multipliers, frequency mixers, frequency converters (OPOs, etc.) or spectrally broadening elements such as self-phase modulation or cross-phase modulation materials, photonic crystals, photonic crystal fibers, supercontinuum-producing fibers (photonic, microstructured, suitably doped or with natural negative group velocity dispersion) are arranged.
  • OPOs frequency converters
  • spectrally broadening elements such as self-phase modulation or cross-phase modulation materials, photonic crystals, photonic crystal fibers, supercontinuum-producing fibers (photonic
  • the laser light source for use in confocal microscopy wherein the laser light source is a so-called supercontinuum laser is used in the picosecond pulses considerable energy (15 to 20 W cw power at 80 mHz repetition rate) in a photonic crystal fiber for white light generation are coupled.
  • both the amplifier units of the laser and the supercontinuum fiber age relatively quickly.
  • the laser light source is split up into a permanently operated seed laser and downstream into a two-stage amplifier unit.
  • the seed laser is controlled by its own monitor photodiodes.
  • the first stage of the amplifier unit is continuously operated in accordance with the above statements, while the second amplifier stage, namely the actual power level, is energized and regulated via the pump diodes, for example when pressing a scan button of the confocal microscope.
  • both the lifetime of the power amplifier and the optical fiber, in particular a supercontinuum can be increased considerably, since usually the actual scan time in a conventional confocal microscope in the range of less than 5% to a maximum of 20% of the total period of use.
  • sufficiently stable operation is possible even after less than 1/10 second, so that the user of the confocal microscope does not perceive the "Lifetime Save Mode" realized here, or at least not disturbing it.
  • the method and the device according to the invention are not only used in confocal microscopes, but generally in systems which, in comparison to the
  • FIG. 1 shows a schematic view of the arrangement and the beam path in a laser scanning microscope, in which the device according to the invention uses the method according to the invention and
  • Fig. 2 is a schematic view of the basic structure of a device according to the invention for use of the method according to the invention in
  • Beam path of a laser scanning microscope. 1 shows the example of a laser scanning microscope 1, the use of a device according to the invention and the application of a method according to the invention.
  • Illumination light is emitted by a laser light source 2 and passes through a beam converter 3 into the illumination beam path 4 of the laser scanning microscope 1.
  • the illumination light passes focused through a lens 7 to the sample 8.
  • the light returning from the sample 8 (predominantly
  • Detection light passes through the lens 7 and passes through the scanning device 6 and the beam splitter 5 and passes through a lens assembly 9 and through an aperture 10 to the detector 11th
  • FIG. 2 shows an exemplary embodiment of a device according to the invention for illuminating or irradiating the sample 8 in the case of the laser scanning microscope 1 shown in FIG. 1.
  • the laser light source 2 is switched to a trigger signal immediately before the actual need, i. immediately before the image acquisition, quickly turned on.
  • the laser light source 2 comprises a seed laser 12 which is permanent operated or energized.
  • the actual laser amplifier is constructed in two stages, namely comprises a preamplifier 14 and a power amplifier 15. Both the preamplifier 14 and the power amplifier 15 is a pump source 16, 17 assigned to namely the preamplifier 14 and the power amplifier 15 to energize or fast to activate ,
  • the preamplifier 14 and the seed laser 12 are operated permanently. This means that the pump sources 13 and 16 permanently energize the seed laser 12 and the preamplifier 14.
  • the pump sources 13, 16 and 17 are controlled or acted upon by a control device 18.
  • the control device 18 comprises a control loop which regulates the current of the respective pump source 13, 16 and 17 as a function of the output power of the laser light source 2.
  • a photodiode 19 or an array of corresponding photodiodes is provided which is preferably designed as a monitor photodiode (s) is (are).
  • FIG. 2 further shows that the laser light source 2 is utilized by suitable means 20 as supercontinuum laser.
  • This means 20 the photodiode 19 is arranged downstream for power measurement. Determined performance data are supplied to the control device 18.
  • the laser light is selected via an AOTF 21 in the wavelength and coupled into the beam path of the laser scanning microscope 1 ⁇ . From there, a trigger line 22 leads to the control device 18, which in turn acts on the pump sources 13, 16 and 17.

Abstract

La présente invention concerne un procédé et un dispositif d'éclairage ou d'irradiation d'un objet, d'un échantillon (8), ou analogue, aux fins de prise de vue ou d'analyse, en particulier pour utilisation dans un microscope à laser (1), de préférence dans un microscope confocal pourvu d'une source de lumière laser (2) émettant la lumière d'éclairage. Le couplage de la lumière laser dans la trajectoire du faisceau d'éclairage (4) se fait directement ou par l'intermédiaire d'une fibre optique. L'invention se caractérise en ce que la source de lumière laser (2) se commute rapidement au signal de déclenchement, immédiatement avant qu'il y en ait effectivement besoin, par exemple immédiatement avant la prise de vue.
EP08787083A 2007-08-21 2008-08-11 Procédé et dispositif d'éclairage d'un échantillon dans un microscope laser Ceased EP2181350A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE102007039498.7A DE102007039498B4 (de) 2007-08-21 2007-08-21 Verfahren und Vorrichtung zur Beleuchtung und/oder Bestrahlung eines Objekts oder einer Probe
PCT/EP2008/060508 WO2009024490A2 (fr) 2007-08-21 2008-08-11 Procédé et dispositif d'éclairage ou d'irradiation d'un objet, d'un échantillon, ou analogue

Publications (1)

Publication Number Publication Date
EP2181350A2 true EP2181350A2 (fr) 2010-05-05

Family

ID=39791128

Family Applications (1)

Application Number Title Priority Date Filing Date
EP08787083A Ceased EP2181350A2 (fr) 2007-08-21 2008-08-11 Procédé et dispositif d'éclairage d'un échantillon dans un microscope laser

Country Status (4)

Country Link
US (1) US8913317B2 (fr)
EP (1) EP2181350A2 (fr)
DE (1) DE102007039498B4 (fr)
WO (1) WO2009024490A2 (fr)

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE102012016410B9 (de) 2012-08-21 2020-01-09 Toptica Photonics Ag Konfokalmikroskop
JP6378869B2 (ja) * 2013-10-24 2018-08-22 株式会社キーエンス 顕微鏡、顕微鏡システム、制御方法およびプログラム
CN107678104A (zh) * 2016-08-02 2018-02-09 南京理工大学 一种指示激光器耦合装置及其调整方法

Family Cites Families (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3782823A (en) * 1972-03-23 1974-01-01 American Optical Corp Laser microprobe
US4555620A (en) * 1983-01-11 1985-11-26 Bausch & Lomb Incorporated Automatic illumination control for multi-objective optical instruments such as microscopes
YU208988A (en) * 1988-11-10 1990-12-31 Inst Jozef Stefan Device for rasters making
DE69028687T2 (de) * 1989-12-15 1997-04-10 Canon Kk Vorrichtung zur optischen Messung einer Probe
CA2062106C (fr) * 1991-12-23 2002-08-20 Gregg Marwin Detecteur d'objets pour lecteur de codes a barres a laser
GB2290411A (en) * 1994-06-15 1995-12-20 Zeiss Stiftung Laser and adapter for mounting it on a surgical microscope
US5757831A (en) * 1996-07-12 1998-05-26 Lightwave Electronics Corp. Electronic suppression of optical feedback instabilities in a solid-state laser
DE10115577A1 (de) 2000-06-17 2001-12-20 Leica Microsystems Scanmikroskop mit mehrbandiger Beleuchtung und optisches Bauteil für ein Scanmikroskop mit mehrbandiger Beleuchtung
JP4576112B2 (ja) * 2003-11-19 2010-11-04 オリンパス株式会社 共焦点レーザ顕微鏡
DE102004051548A1 (de) * 2004-10-20 2006-05-04 Carl Zeiss Jena Gmbh Beleuchtungseinrichtung für Mikroskope
TWI261959B (en) * 2005-06-24 2006-09-11 Leadlight Technology Inc Power control device of laser module and method thereof
US7391561B2 (en) * 2005-07-29 2008-06-24 Aculight Corporation Fiber- or rod-based optical source featuring a large-core, rare-earth-doped photonic-crystal device for generation of high-power pulsed radiation and method

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO2009024490A2 *

Also Published As

Publication number Publication date
DE102007039498B4 (de) 2017-08-03
US20110279890A1 (en) 2011-11-17
US8913317B2 (en) 2014-12-16
DE102007039498A1 (de) 2009-02-26
WO2009024490A2 (fr) 2009-02-26
WO2009024490A3 (fr) 2009-05-07

Similar Documents

Publication Publication Date Title
DE102008018476B4 (de) Mikroskopievorrichtung
EP1108227B1 (fr) Mode de fonctionnement d'un microscope confocal, a balayage laser
DE10235914B4 (de) Lichtquelle zur Beleuchtung mikroskopischer Objekte und Scanmikroskopsystem
EP1186882B1 (fr) Procédé et dispositif pour la détection de lumière fluorescente dans la microscopie confocale à balayage
EP2761276B1 (fr) Circuit d'évaluation pour un détecteur optoélectronique et procédé d'enregistrement d'événements de fluorescence
EP2535756A1 (fr) Microscope et procédé de microscopie à fluorescence imagée
DE10251888A1 (de) Treiber für Pockelzelle und Verwendung der Pockelzelle in Lasersystemen
DE102004009068B4 (de) Faseroptische Verstärkung von Lichtimpulsen
DE10139754B4 (de) Beleuchtungsverfahren für ein Scanmikroskop und Scanmikroskop
DE102004017956A1 (de) Mikroskop zur Untersuchung der Lebensdauer angeregter Zustände in einer Probe
DE102007039498B4 (de) Verfahren und Vorrichtung zur Beleuchtung und/oder Bestrahlung eines Objekts oder einer Probe
DE102011122230B4 (de) Optikanordnung und Verfahren zum Untersuchen oder Bearbeiten eines Objekts
DE10029167B4 (de) Verfahren und Vorrichtung zur Temperaturstabilisierung optischer Bauteile
DE10033269B4 (de) Vorrichtung zum Einkoppeln von Licht mindestens einer Wellenlänge einer Laserlichtquelle in ein konfokales Rastermikroskop
WO2014095392A2 (fr) Système laser à courtes impulsions
DE102011086230A1 (de) Verfahren und Vorrichtung zur Beleuchtung und Detektion in der RESOLFT-Mikroskopie
WO2008086996A1 (fr) Dispositif d'éclairage à éléments optiques non linéaires pour produire une lumière laser dans un large domaine spectral avec une densité de puissance spectrale homogène
DE102014010185A1 (de) Verfahren zum Betrieb eines Laser-Scanning-Mikroskops
DE102013114580B4 (de) Lichtquellenvorrichtung
DE102008005337B4 (de) Laser-Scanning-Mikroskop mit variabler Lichtintensität und Steuerverfahren für ein solches
DE202013006817U1 (de) Gepulste Laserlichtquelle für die Fluoreszenzanregung
DE102009021993B4 (de) Scanmikroskop und zugehörige Verfahren
DE10313138A1 (de) Vorrichtung zur Beleuchtung eines Objekts
DE10220871A1 (de) Optisch-Parametrischer Oszillator und Verstärker
DE102007009659B4 (de) Verwendung eines diodengepumpten Festkörperlasers und Laser-Scanning-Mikroskop mit einem UV-Beleuchtungsstrahlengang

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

17P Request for examination filed

Effective date: 20100318

AK Designated contracting states

Kind code of ref document: A2

Designated state(s): AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MT NL NO PL PT RO SE SI SK TR

AX Request for extension of the european patent

Extension state: AL BA MK RS

17Q First examination report despatched

Effective date: 20100705

REG Reference to a national code

Ref country code: DE

Ref legal event code: R003

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE APPLICATION HAS BEEN REFUSED

18R Application refused

Effective date: 20120117